B Y T

TRNG I HC DC H NI
**********

KHNG TRNG QUN

NGHIN CU TC DNG GII LO U

THC NGHIM CA STEPHANIA SINICA
DIELS. V STEPHANIA DIELSIANA Y.C. WU,
H TIT D MENISPERMACEAE

KHA LUN TT NGHIP DC S

H NI - 2013

B Y T
TRNG I HC DC H NI

KHNG TRNG QUN

NGHIN CU TC DNG GII LO U

THC NGHIM CA STEPHANIA SINICA
DIELS. V STEPHANIA DIELSIANA Y.C. WU,
H TIT D MENISPERMACEAE

KHA LUN TT NGHIP DC S

Ngi hng dn:

1.TS. o Th Vui
2.TS. Nguyn Hong Anh
Ni thc hin:
B mn Dc lc

H NI 2013

LI CM N
Bng tt c s chn thnh v lng bit n su sc, em xin gi li cm n ti
TS. Nguyn Hong Anh, TS. o Th Vui, Ths. Nguyn Thu Hng, B mn
Dc lc, TS. Nguyn Qunh Chi, B mn Dc liu, TS. Nguyn Quc Huy,
B mn Thc vt, trng H Dc H Ni.
L nhng ngi thy, ngi c trc tip hng dn, tn tnh ch bo v
gip em trong sut qu trnh thc hin v hon thnh kha lun.
Em xin gi li cm n ti anh inh i , k thut vin ti b mn Dc
lc trng H Dc H Ni nhit tnh gip v trc tip thc hin nhiu cng
vic xuyn sut trong qu trnh tin hnh ti.
Nhn y em cng xin gi li cm n ti:
Cc thy c, cc anh ch k thut vin trong b mn Dc lc to mi
iu kin, gip em c th hon thnh kha lun mt cch tt nht.
Cc thy c trong Ban gim hiu nh trng, cc phng ban, cc b mn
trng i hc Dc H Ni tn tnh dy d, gip em trong qu trnh hc tp
ti trng.
Em cng xin by t lng cm n chn thnh ti nhng ngi thn trong gia
nh, nhng ngi bn lun kp thi ng vin, ng h em trong sut qu trnh
hc tp ti trng cng nh trong thi gian thc hin kha lun.
Em xin chn thnh cm n.

H Ni, thng 5 nm 2013

Sinh vin
Khng Trng Qun

MC LC
Trang
DANH MC CC K HIU, CH VIT TT
DANH MC CC BNG
DANH MC CC HNH
T VN ........................................................................................................1
CHNG I. TNG QUAN ..................................................................................3
1.1. Chi Stephania Lour.: Thnh phn ha hc v cc tc dng dc l tm
thn/thn kinh ....................................................................................................3
1.1.1.Thnh phn ha hc .................................................................................3
1.1.2.Hm lng alcaloid ton phn v L-tetrahydropalmatin trong cc loi bnh
vi Vit Nam .................................................................................................3
1.1.2.1.Alcaloid ton phn .............................................................................3
1.1.2.2.Hm lng L-tetrahydropalmatin (Rotundin).....................................4
1.1.3.Tc dng dc l hng thn kinh ca cc loi thuc chi Stephania Lour.
.........................................................................................................................6
1.2. V hai loi Stephania dielsiana Y.C.Wu v Stephania sinica Diels............9
1.2.1.c im thc vt ....................................................................................9
1.2.2.Thnh phn ha hc ...............................................................................10
1.2.3.Cc nghin cu v tc dng dc l hng thn kinh v tm thn..........14
CHNG II. NGUYN LIU V PHNG PHP NGHIN CU............. 15
2.1.Nguyn liu .................................................................................................15
2.1.1.Dc liu ............................................................................................... 15
2.1.2.Ha cht v thit b ................................................................................15
2.1.3.ng vt thc nghim ............................................................................16
2.1.4.Chun b thuc th .................................................................................16
2.2.Phng php nghin cu ...........................................................................17

2.2.1.S thit k nghin cu .......................................................................17

2.2.2.Ni dung nghin cu ..............................................................................17
2.2.3.iu kin tin hnh th nghim............................................................... 18
2.2.4.Liu s dng trong nghin cu ............................................................... 18
2.2.5.M hnh gy stress cho chut bng phng php nui c lp .................19
2.2.6.nh gi tc dng gii lo u trn m hnh EPM .....................................19
2.2.7.nh gi tc dng gii lo u trn m hnh ETM .....................................21
2.2.8.nh hng trn thi gian ng do thiopental. ..........................................22
2.3.X l s liu ................................................................................................ 23
CHNG III. KT QU V BN LUN ....................................................... 24
3.1.nh gi tc dng gii lo u ca dch chit nc ton phn S. dielsiana v
S. sinica .............................................................................................................24
3.1.1.Trn m hnh EPM chut khng chu stress ........................................24
3.1.2.Trn m hnh EPM chut chu stress do c lp....................................26
3.1.3.Trn m hnh ETM ................................................................................28
3.2. nh gi tc dng an thn ca dch chit nc ton phn ca S. dielsiana
v S. sinica ........................................................................................................29
3.2.1.Trn chut khng chu stress ..................................................................29
3.2.2.Trn chut chu stress do c lp ............................................................. 30
3.3. nh gi tc dng gii lo u v an thn ca alcaloid ton phn t S.
dielsiana v S. sinica .........................................................................................31
3.3.1. Chit xut alcaloid t c 2 loi Stephania..............................................31
3.3.2.nh gi tc dng gii lo u ca alcaloid ton phn t S. dielsiana v S.
sinica ..............................................................................................................32
3.3.3.nh gi tc dng an thn ca alcaloid ton phn t 2 loi Stephania ....35
3.4.Bn lun......................................................................................................36
KT LUN V XUT ................................................................................. 41
TI LIU THAM KHO

DANH MC CC K HIU, CH VIT TT

Akt

Protein kinase B

BDNF

Yu t kch thch thn kinh n t no (Brain-derived

neurotrophic factor)

CREB

Protein lin kt tng phn p ng vi AMP vng (Cyclic-AMP

response element-binding protein)

CRF

Yu t gii phng corticotropin (Corticotropin-releasing factor)

DZP

Diazepam

EPM

Ch thp nng cao (Elevated plus maze)

ERK

Protein chuyn gc phosphat iu ha tn hiu ngoi bo

(Extracellular signal-regulated kinase)

ETM

Ch T nng cao (Elevated T maze)

GABA

Gamma amino butyric acid

GAD

Ri lon lo u lan ta (Generalized anxiety disorder)

IL-1

Interleukin 1 beta

NF-B

Yu t nhn lm giu chui nh kappa ca t bo B hot ha

(Nuclear factor kappa-light-chain-enhancer of activated B cells)

NMDA

N-methyl-D-aspartat

PD

Ri lon hong s (Panic disorder)

SPD

Stepholidin

THP

Tetrahydropalmatin

Rf

H s lu

MeOH

Methanol

DANH MC CC BNG
Bng

Tn bng

Trang

1.1

Kt qu nh lng alcaloid ton phn (Nguyn Tin Vng)

1.2

Kt qu nh lng alcaloid ton phn (L nh Mi)

1.3

Kt qu nh lng THP (Nguyn Tin Vng)

1.4

Kt qu nh lng THP (L nh Mi)

1.5

Kt qu nh lng THP (Nguyn Quc Huy)

3.1

3.2
3.3
3.4

3.5

3.6

3.7

Tc dng ca dch chit nc ton phn v diazepam trn thi gian

lu v s ln ra tay m ca chut khng chu stress trn m hnh
EPM
Tc dng ca dch chit nc ton phn v diazepam trn s ln
vo tay kn v tng s ln lu ti tay kn, tay m ca chut khng
chu stress trn m hnh EPM
Tc dng ca dch chit nc ton phn v diazepam trn s ln v
thi gian lu ti tay m ca chut chu stress trn m hnh EPM
Tc dng ca dch chit nc ton phn v diazepam trn s ln
vo tay kn v tng s ln lu ti tay kn, tay m ca chut chu
stress trn m hnh EPM
Tc dng ca dch chit nc ton phn v clomipramin trn chut
khng chu stress th hin qua cc ch tiu nh gi trn m hnh
ETM
Tc dng ca alcaloid ton phn, diazepam v tetrahydropalmatin
trn s ln v thi gian lu ti tay m ca chut khng chu stress
trn m hnh EPM
Tc dng ca alcaloid ton phn, diazepam v tetrahydropalmatin
trn s ln vo tay kn v thi gian lu ti tay kn, tay m ca chut
khng chu stress trn m hnh EPM

24

25
27
27

29

33

34

DANH MC CC HNH
Hnh

Tn hnh

Trang

1.1

Sc k ca dch chit alcaloid ton phn ca 6 loi thuc chi

Stephania Lour. (UV 254 v 365 nm)

2.1

Hnh nh c loi Stephania sinica Diels.

15

2.2

Hnh nh c loi Stephania dielsiana Y.C.Wu.

15

2.3

Hnh nh chut nui c lp gy stress

19

2.4

M hnh ch thp nng cao

20

2.5

M hnh ch T nng cao

21

2.6

Dng c ch T nng cao

21

3.1

3.2
3.3

3.4

nh hng trn thi gian ng do thiopental ca diazepam (DZP) v

dch chit nc ton phn (DC) ca 2 loi Stephania trn chut
bnh thng
nh hng trn thi gian ng do thiopental ca diazepam (DZP) v
dch chit nc ton phn (DC) ca 2 loi Stephania trn chut chu
stress do c lp
Sc k lp mng phun thuc th hin mu c i chiu vi cht
chun tetrahydropalmatin
nh hng trn thi gian ng do thiopental ca diazepam (DZP),
tetrahydropalmatin (THP) v alcaloid ton phn ca 2 loi
Stephania trn chut bnh thng

30

31
32
35

T VN
Ri lon lo u (anxiety disorders) l s lo s qu mc trc mt tnh hung
xy ra, c tnh cht v l, lp li v ko di gy nh hng n s thch nghi vi
cuc sng ca ngi bnh. y l mt trong nhng ri lon tm l c tnh ph bin
cao, c xu hng ngy cng gia tng, thng kt hp vi nhiu ri lon khc nh
mt ng, trm cm, ri lon n ung, ri lon hnh vi [14]. Hin nay, phng php
iu tr ri lon lo u ch yu l phi hp liu php tm l vi thuc gii lo u
(anxiolytics). Cc thuc gii lo u c ngun gc ha dc chnh c s dng l
cc dn cht ca benzodiazepin, buspiron v cc thuc chng trm cm c ch chn
lc ti thu hi serotonin hoc chng trm cm 3 vng [17]. Hn ch ln nht ca cc
thuc c ngun gc ha dc l tc dng ph, kh nng l thuc thuc v phn ng
cai thuc xy ra khi dng iu tr. Thm vo , gi thnh cao ca cc thuc ny
cng l vn cn quan tm khi s dng thuc hng ngy trong mt thi gian di
cho bnh nhn. V vy, s dng cc thuc v sn phm c ngun gc dc liu hin
c coi l hng tip cn b sung v thay th cho cc thuc c ngun gc ha
dc trong iu tr ri lon lo u [47].
Chi Stephania Lour. l mt trong nhng chi ln nht thuc h Tit d
(Menispermaceae) vi khong 60 loi, phn b ch yu vng nhit i v cn
nhit i ca chu , chu Phi v chu c. Trong lnh vc dc l thn kinh v tm
thn, nghin cu v cc loi trong chi ch yu tp trung trn tc dng chng lon
tm thn, hng iu tr Alzheimer, an thn v gii lo u. Nhiu nghin cu ha hc
v dc l c thc hin lm sng t cc hng tc dng trn trong nhiu
alkaloid c hot tnh sinh hc c phn lp v xc nh cu trc [48].
Ti Vit nam, cc loi ca chi Stephania Lour. c nghin cu kh y
v ha hc v tc dng sinh hc hng an thn trn c s chit tch Ltetrahydropalmatin v kinh nghim s dng ca nhn dn [6], [16]. Hng nghin
cu ca ti tp trung vo nh gi tc dng dc l thn kinh v tm thn ca hai
loi Stephania c pht hin Ba v (H ni) v Phong Nha (Qung Bnh) l
Stephania sinica Diels. (bnh vi tn ngn) v Stephania dielsiana Y.C. Wu. (c

dm) [6]. Quan st ca Phm Thanh K, Nguyn Quc Huy cho thy dch chit S.
dielsiana c tc dng gim hot ng ca chut v ko di thi gian ng do
thiopental gp 3 ln v 8 ln so vi nhm chng khi dng liu tng ng 1,25g/kg
v 2,5g/kg [10]. Nghin cu ban u v ha hc ca Quyn, Nguyn Quc Huy
v cng s cng cho thy c s khc bit v thnh phn alcaloid gia cc loi
Stephania, trong khi t l L-tetrahydropalmatin S. sinica l khong 2,43% th S.
dielsiana ch l 0,4% [6]. Nhng kt qu ny gi cho hng nghin cu su hn
v tc dng an thn v gii lo u ca S. sinica v S. dielsiana ng thi tm hiu
xem liu tc dng ny c ph thuc vo s c mt ca L-tetrahydropalmatin trong
dc liu hay khng.
Xut pht t thc t , ti c trin khai vi mc tiu nh gi tc dng
gii lo u v an thn thc nghim ca dch chit nc v alcaloid ton phn ca
Stephania sinica Diels. v Stephania dielsiana Y.C. Wu. trn chut bnh thng v
chut chu stress do c lp.

CHNG I. TNG QUAN

1.1. Chi Stephania Lour.: Thnh phn ha hc v cc tc dng dc l tm
thn/thn kinh
1.1.1. Thnh phn ha hc
Thnh phn ha hc ca chi Stephania Lour. gm c alcaloid, flavonoid, tinh
bt, ng kh, acid malic, enzym oxydase....Trong alcaloid l thnh phn chnh
v c quan tm nhiu nht [48]. Cc alcaloid phn lp t cc loi trong chi
Stephania Lour. c th xp vo 9 nhm: benzylisoquinolin, bisbenzylisoquinolin,
aporphin, proaporphin, protoberberin, morphinan, hasubanan, stephaoxocan v
eribidin [6]. Trong s ny L-tetrahydropalmatin (THP) l alcaloid c nghin cu
nhiu nht pht trin ng dng sn xut thuc an thn, gy ng.
1.1.2. Hm lng alcaloid ton phn v L-tetrahydropalmatin trong cc loi bnh
vi Vit Nam
1.1.2.1. Alcaloid ton phn
Nm 2000, Nguyn Tin Vng cng b hm lng alcaloid ton phn t
3 loi bnh vi nghin cu nh sau :
Bng 1.1: Kt qu nh lng alcaloid ton phn (Nguyn Tin Vng)
Stt

Loi

Ni thu hi

Hm lng alcaloid ton phn (%)

S. glabra(Roxb.) Miers

Ninh Bnh

2,96

S. kuinanensis H.S.Lo

Lng Sn

4,41

Stephania sp3.

Qung Ninh

2,32

Nm 2000, L nh Mi kho st hm lng alcaloid ton phn trong 5

loi bnh vi nghin cu :

Bng 1.2: Kt qu nh lng alcaloid ton phn (L nh Mi)

Hm lng alcaloid
Stt

Loi

Ni thu hi

S. brachyandra Diels.

Sapa-Lo Cai

10,8

S. glabra (Roxb.) Miers

Mai Chu-Ha Bnh

3,4

S. sinica Diels.

Tam ip-Ninh Bnh

3,6

S. kwangsiensis H.S.Lo

Cm Ph-Qung Ninh

3,5

S. hainanensis H.S.Lo

H Nam, Ninh Bnh

2,6

ton phn (%)

Nm 2003, V Xun Giang nh lng alcaloid ton phn loi

S.viridiflavens H.S. Lo thu hi Sn La l 2,92% (trch theo Nguyn Quc Huy
[6]). Nm 2004, Phng Loan xc nh hm lng alcaloid ton phn trong
c loi S. brachyandra Diels. l 10,38%.
1.1.2.2. Hm lng L-tetrahydropalmatin (Rotundin)
Nm 2000, Nguyn Tin Vng xc nh hm lng THP t 3 loi bnh vi
nghin cu :
Bng 1.3: Kt qu nh lng THP (Nguyn Tin Vng)
Stt

Loi

Hm lng THP (%)

S. glabra (Roxb.) Miers

0,59

S. kuinanensis H.S.Lo

3,06

Stephania sp3.

1,52

Nm 2000, L nh Mi kho st hm lng THP trong 5 loi bnh vi

nghin cu :

Bng 1.4: Kt qu nh lng THP (L nh Mi)

Hm lng
Hm lng
Stt

Loi

S. brachyandra Diels.

THP (%)

THP/alcaloid ton
phn (%)

5,1

50,1

2,1

50,0

S. glabra (Roxb.)
2

Miers

S.sinica Diels.

1,4

46,4

S.kwangsiensis H.S.Lo

1,8

50,5

S.hainanensis H.S.Lo

0,6

18,3

V Xun Giang xc nh hm lng THP v palmatin trong loi S.

viridiflavens H.S. Lo thu hi Sn La l: 0,63% v 0,32% . Phm Th Duyn (2004)
nghin cu nh lng THP bng phng php sc k lng hiu nng cao (HPLC)
cho thy hm lng THP trong loi S. dielsiana (thu hi Ba V-H Ty), loi S.
viridaflavens (thu hi Sn La) v Stephania sp. (trong vin Balok) ln lt l:
0,40%; 0,62% v 1,63%. Phng Loan xc nh hm lng THP ca loi S.
brachyandra Diels. c l 3,7% v thn l 0,67%.
Nm 2010, Nguyn Quc Huy tin hnh nh lng THP trong 3 loi
nghin cu [6]:
Bng 1.5: Kt qu nh lng THP (Nguyn Quc Huy)
Stt

Loi

Hm lng THP (%)

S. brachyandra Diels.

3,69

S. dielsiana Y. C. Wu.

0,40

S. sinica Diels.

2,43

C th thy r c s khc bit rt ln v hm lng alcaloid v THP gia cc

loi chi Stephania Lour. pht hin ti Vit Nam. Kt qu kho st trn sc k lp
mng ca Quyn v Nguyn Quc Huy cng th hin r iu ny

Hnh1.1.Sc k ca dch chit alcaloid ton phn ca 6 loi thuc chi Stephania
Lour. (UV 254 v 365 nm)
BR: S. brachyandra, GL: S. glabra, KW: S. kwangsiensis, SI: S. sinica; SP1:
Stephania sp.1 (Mu Ha Bnh), DI: S. dielsiana, R: L-tetrahydropalmatin
Dung mi trin khai sc k: Cloroform : Aceton : NH4OH [8 : 2 : 0,1]
1.1.3. Tc dng dc l hng thn kinh ca cc loi thuc chi Stephania Lour.
Trong lnh vc dc l thn kinh v tm thn, nghin cu v cc loi trong
chi Stephania ch yu tp trung trn tc dng chng ri lon tm thn, tc dng
hng iu tr Alzheimer v tc dng an thn v gii lo u.
(-)- Stepholidine (SPD) phn lp t S. intermedia H.S. Lo c chng
minh c hot tnh i khng receptor D2 ca dopamin vng bng mi (ventral
tegmental area) v nhn vng (nucleus accumbens), kch thch hot ng ca
tyrosinhydroxylase, iu ha gii phng dopamin v c hiu qu chng lon thn
trn m hnh gy tm thn phn lit ng vt thc nghim [52]. Trn lm sng,
gip ci thin c triu chng m tnh v dng tnh bnh nhn tm thn phn lit
ng thi t gy tc dng ngoi thp [52]. Bi-acetylated l-stepholidine, mt dn cht

c sinh kh dng tt hn SPD, vi c tnh ch vn trn receptor D1 v receptor 5HT1A, i khng trn receptor D2, lm gim kch thch v ci thin chc nng vn
ng, nhn thc chut cng trong m hnh gy tm thn phn lit thc nghim bi
phencyclidin [25].
Tetrahydropalmatin (THP), mt alcaloid c tm thy hu ht cc loi
thuc chi Stephania, c i lc gn cao vi receptor 1A v 2A-adrenergic, D1, D2 v
D3 ca dopamin, receptor 5-HT1A, 5-HT1D, 5-HT4 v 5-HT7 ca serotonin [34]. Tc
dng i khng ca THP trn receptor dopamin gim dn theo th t D1, D2, D3.
THP c tc dng i khng receptor 1-adrenergic, ch vn trn receptor 2adrenergic v l cht iu ha d lp th dng tnh ca receptor GABAA [26], [51].
THP i khng tc dng hot ha cc receptor serotonin, noradrenalin v dopamin
ca cocain, ng thi tc dng trn receptor 5-HT1A v -adrenergic gip gim nh
tc dng ngoi thp [37]. THP c tc dng gim cc triu chng trong qu trnh cai
nghin cocain v heroin trn thc nghim chut cng [53]. THP c ch s ph
thuc tm thn gy ra bi oxycodon thng qua nh hng ti s phosphoryl ha
CREB (cyclic-AMP response element-binding protein) v ERK (Extracellular
signal-regulated kinases) nhn vng v hi hi m (hippocampus) chut cng
[35].
Theo hng tip cn sng lc dc liu c tc dng c ch
acetylcholinesterase c tim nng trong iu tr bnh Alzheimer, dch chit c loi
S. suberosa Forman. nng 0,1 mg/ml c ch trn 90% hot tnh
actetylcholinesterase [29]. Ba alkaloid protoberberin (stepharanine, cyclanoline v
N-methyl stepholidine) phn lp t loi S. venosa (Blume) Spreng. c hot tnh c
ch mnh acetylcholinesterase (tim nng trong iu tr sa st tr tu v bnh
Alzheimer) vi gi tr IC50 tng ng l 14,1; 9, 23 v 31,3 M [28]. Tetrandrin
c chit xut v phn lp t loi S. tetrandrae S. Moore c ch hot ng ca
NF-B, gim ng k s to thnh cc yu t gy vim IL-1 v TNF-, bo v t
bo thn kinh khi c tnh gy ra bi amyloid- protein, ci thin tnh trng sa st
tr nh v gim kh nng hc nh trn chut cng (m hnh m cung nc Morris),

to tim nng trong iu tr bnh Alzheimer [27]. Nh c tnh ch vn trn

receptor D1 v i khng trn receptor D3, SPD lm gim nh ri lon vn ng
bnh nhn mc bnh Parkinson [52].
Dch chit nc ca 3 loi S. baicalensis, S. tetrandra, S. miltiorrhiza c ch
receptor N-methyl-D-aspartat (NMDA), c tc dng bo v t bo thn kinh to
tim nng trong iu tr t qu. Nghin cu su hn v tc dng dc l v c ch
tc dng ca mt s alcaloid cho thy stepholidin (SPD) thng qua con ng tn
hiu PI3-K/Akt lm tng nng Akt phosphoryl ha (pAkt), c ch apoptosis, bo
v t bo thn kinh khi phi nhim vi H2O2 [55].
Tetrandrin, mt alkaloid nhm bisbenzyllisoquinoline phn lp t S.
tetrandrae S. Moore cng c chng minh c tc dng an thn th hin trn
kh nng ko di thi gian ng do pentobarbital trn chut nht vi c ch thng
qua nh hng trn dn truyn serotonin h thn kinh trung ng [58]. Dch chit
S. venosa liu 20mg/kg dng ng ung tng s ln ra tay m, tng thi gian lu
ti tay m trn chut cng, th hin tc dng gii lo u. Mc liu ny khng nh
hng ti thi gian bt ng trong th nghim chut bi cng bc lin quan n
tc dng chng trm cm [40]. Tetrandrin o ngc tc dng lm gim hm lng
serotonin v noradrenalin hi hi m gy ra bi reserpin, th hin hot tnh chng
trm cm thng qua nh hng trn dn truyn cc amin giao cm v tng nng
BDNF (brain-derived

neurotrophic

factor). Hot tnh chng trm cm ca

tetrandrin c th hin thng qua vic lm gim thi gian bt ng ca chut nht
2 m hnh th nghim chut bi cng bc v test treo ui [23].
Tc dng c ch thn kinh trung ng ca dl-tetrahydropalmatin (THP), mt
alkaloid c phn lp t nhiu loi trong chi Stephania c chng minh trn
cc m hnh dc l thc nghim. liu thp (0,5 10 mg/kg), THP c tc dng
gii lo u r rt, tc dng an thn v gin c ch th hin liu cao (50 mg/kg). Tc
dng gii lo u ca tetrahydropalmatin b i khng bi flumazenil, mt cht cnh
tranh v tr gn ca benzodiazepin trn GABAA receptor, chng t c ch gii lo u

ca THP thng qua, t nht mt phn, tng tc vi v tr gn ca benzodiazepin

trn receptor GABAA [26], [32]. Nguyn Tin Vng tin hnh thm d tc dng
ca l-THP trn in no th. Tt c cc th c tin hnh t in cc vo vng
cm gic, vn ng v th li thn no. Sau khi cho th ung l-THP vi liu
100mg/kg th trng trong 7 ngy, thnh phn cc sng in no trong v no vng
cm gic-vn ng v trong th li thn v no c nhng bin ng r. S tng
thnh phn sng chm delta v gim thnh phn sng nhanh beta chng t rng lTHP c tc dng tng cng c ch t bo thn kinh v bn cu i no v th
li thn no [16]. Phm Duy Mai v cng s cng nghin cu tc dng an thn
ca THP, kt qu cho thy gindarin hydroclorid (l-THP hydroclorid) mc liu
30mg/kg v 100mg/kg c tc dng ko di thi gian ng gp do thiopental gp 1.5
ln so vi nhm chng (trch theo Nguyn Quc Huy [6]). Cng theo Nguyn Tin
Vng, l-THP liu 40mg/kg ko di thi gian ng do pentobarbital gp 8 ln so vi
nhm chng [16].
1.2. V hai loi Stephania dielsiana Y.C.Wu v Stephania sinica Diels.
1.2.1. c im thc vt
Stephania dielsiana Y.C.Wu.: Tn thng dng: C dm. Tn khc: Bnh
vi nha tm, c tm (Ty, Tuyn Quang), c g p. Phn b: Lo Cai, Yn Bi,
Bc Cn, Thi Nguyn, Sn La, Ha Bnh, Ph Th, H Ty, Bc Giang, Bc Ninh,
Qung Ninh. Dy leo nh, sng nhiu nm. R c to. Thn leo cun di khong 3m.
Thn non mu tm hng nht. Ngn non, cung l v cung cm hoa c mu tm
hng. Hoa n tnh khc gc. Cm hoa c do 3-5 xim nh hp thnh xim tn kp.
Hoa nh, c cung ngn; i 6, mu tm, xp 2 vng, cnh hoa 3, hnh qut trn,
mu vng cam. Ct nh ngn, bao phn 6, dnh thnh a 6 . Cm hoa ci gm 7-8
u nh, cung rt ngn, xp thnh dng u. Hoa nh; i 1, mu tm hng; cnh
hoa 2, hnh qut trn, mu vng cam v c cc vn tm; bu hnh trng, u nhy c
4-5 thy dng di. Qu hnh trng o, hi dt. Ht hnh trng ngc ct u, c l
thng gia, trn lng ht c 4 hng gai nhn, cong [6].

10

Stephania sinica Diels.: Tn ng ngha: Stephania rotunda Lour.. Tn

thng dng: Bnh vi, bnh vi Trung Quc. Tn khc: C mt Trung Quc, dy
li tin bc. Phn b: Lng Sn, Qung Ninh, H Ty, Ha Bnh, H Nam, Ninh
Bnh ti B Ra- Vng Tu. Dy leo cun, thn c rnh dc. R c to, mc n c
hoc thnh m. L c phin hnh lng, khng lng, mt di mu nht, cung di
n 30cm. Cm hoa c, xim tn kp. Hoa c: i 6, cnh hoa 3. Cm hoa ci
dng u. Hoa ci: i 1, cnh 2. Cm qu hnh cu. Ht hnh mng nga, dc theo
chiu lng c 4 hng g [6].
1.2.2. Thnh phn ha hc
Theo Nguyn Quc Huy, trong c 2 loi bnh vi nghin cu u cha
alcaloid, tinh bt, ng kh t do, acid hu c, trong alcaloid l thnh phn
chnh [6].
Hm lng alcaloid ton phn l 3,6% [12] vi S. sinica v bin thin t
2,68% n 4,85% vi S. dielsiana ty thuc vo tui ca cy [7]. Trong khi t l Ltetrahydropalmatin S. sinica l 2,430,04% th S. dielsiana ch l 0,400,01%
[6].
Hin phn lp c 5 alcaloid t loi S.sinica, bao gm: ltetrahydropalmatin (1) , runanin (2), cepharanthin (3), corydalmin (4) v sinomenin
(5) [6], [9], [39].
OCH3
H3CO
H3CO

O
+

N
H3C

N
CH3
O

OCH3
OCH3

O
O

O
H3CO

CH3

11

H3CO

H3CO

H3CO

H3CO

H
OCH 3

OCH 3

OCH 3

OH

1
H3CO

HO
N-CH3
O
OCH3

5
Vi S.dielsiana hin c 15 alcaloid c phn lp t c ca loi ny, bao
gm : dehydrostesakin (6), oxocrebanin (7), 1,2 methylen-dioxy- 8 methoxy- 7
oxodibenzoquinolin (8), l-tetrahydropalmatin, oxostephanin (9), dehydrocrebanin
(10), ayuthianin (11), cephamorphinanin (12), aknadinin (13), liriodenin (14),
(-) corydalmin, nor-canelillin (15), sinomenin, stephanin (16), dehydrostephanin
(17), crebanin (18) [6], [11], [21], [56].

12

O
O

HO

N
OCH 3

H3CO

CH3

OCH 3

7
O

O
N

H3C

O
H3CO

OCH3

OCH3

15

H3CO

O
N

H3CO

H3C

OCH3

OCH3

H 3CO

9
O

H3C

CH3
H
OH

H3CO

CH3

OCH 3

10

11

13

H3CO

HO

HO

N-CH3

NH
HO

OCH 3

OCH 3
OCH 3

OH

12

13

H3CO

N
H3CO

H
OCH3

OH

14

4
H3CO

HO
HO

NH
H3CO

N-CH 3

OH
O
OCH 3

15

5
O

H3C

O
N

H3C

H3CO

H 3CO

16

17

14

1.2.3. Cc nghin cu v tc dng dc l hng thn kinh v tm thn

Tm kim trn c s d liu Pubmed/Medline vi cc t kha Stephania
sinica Diels. v Stephania dielsiana Y.C. Wu. cha tm thy bt c cng b no v
tc dng sinh hc lin quan n tc dng an thn v gii lo u ca hai loi cng b
bi cc tc gi nc ngoi.
Trong nc, Nguyn Quc Huy v cng s xc nh c liu LD50 ca
c loi S. dielsiana l 22,2 g/kg v th tc dng chng vim v gim au thc
nghim ca loi ny [8]. Theo , c 2 mc liu 2,5 g/kg v 5 g/kg u khng c tc
dng gim au theo c ch trung ng theo phng php gy au bng mm nng.
Theo hng tip cn sng lc dc liu c tc dng c ch acetylcholinesterase c
tim nng trong iu tr bnh Alzheimer, Quyn v cng s pht hin dch
chit methanol ca Stephania sinica c tc dng c ch mnh nht enzym ny (hot
c ch 85% nng 250 g/ml) ng thi phn lp c 2 alkaloid l ltetrahydropalmatin v sinomenin trong phn on c tc dng [13]. Quan st ban
u ca Phm Thanh K v Nguyn Quc Huy cho thy t mc liu 2,5 g/kg dch
chit dc liu S. dielsiana quan st thy s gim hot ng ca chut. Dch
chit dc liu c tc dng ko di thi gian ng do thiopental gp 3 ln v 8 ln so
vi nhm chng khi dng liu tng ng 1,25 g/kg v 2,5 g/kg [10].

15

CHNG II. NGUYN LIU V PHNG PHP NGHIN CU

2.1. Nguyn liu
2.1.1. Dc liu
C ca loi bnh vi Stephania dielsiana Y.C.Wu., h Tit d
(Menispermaceae) thu hi ti Ba V, H Ni v c ca loi bnh vi Stephania
sinica Diels., h Tit d (Menispermaceae) thu hi ti vn quc gia Phong Nha-K
Bng, Qung Bnh. Mu dc liu c Nguyn Quc Huy, B mn Thc vt,
Trng i hc Dc H Ni nh tn khoa hc v lu gi tiu bn ti phng tiu
bn ca B mn. Dc liu c ra sch, ct nh, phi v sy kh.

Hnh 2.1. Hnh nh c loi S. sinica

Hnh 2.2. Hnh nh c loi S. dielsiana

2.1.2. Ha cht v thit b

Diazepam

(bit

dc

Seduxen

ca

Gedeon

Richter,

Hungari),

tetrahydropalmatin (Rotundin) chun phn tch ca cng ty CP Tm Hiu c

(98,4%), clomipramin (bit dc Clomidep 25 mg ca Cng ty Sun Pharmaceutical
Industries, n ), thiopental natri ca Rotexmedica (c) do cng ty c phn
Dc phm TW2 phn phi.
Dung dch NaCl 0,9%, CMC 0,5%, NH4OH bo ha, cloroform, thuc th
Mayer, thuc th Dragendorf.
Bnh chit Soxhlet, B mn Dc liu , trng H Dc H Ni.
Bm, kim tim nha, ng h bm giy, cn phn tch, cn k thut.

16

Cc c m 100, 250 ml, ng ong 50, 100 ml, chy, ci v mt s dng c

dng trong chn nui v pha ch ca b mn Dc lc, trng H Dc H Ni.
2.1.3. ng vt thc nghim
+ Chut nht trng ging c, chng Swiss albino khe mnh, khi lng
trung bnh 18-22 g c cung cp bi Vin V sinh Dch t Trung ng.
+ Chut cng trng c 2 ging, chng Wistar, khi lng 200-250 g do Hc
vin Qun Y cung cp. Chut c chia l ngu nhin 8-10 con/l, nui trong iu
kin nhit 241oC, nh sng t nhin, c nui bng thc n chun, ung nc
t do.
2.1.4. Chun b thuc th
+ Chit alcaloid: chit bng dung mi hu c trong mi trng kim theo
Dc in Vit Nam IV [3].
Cn chnh xc khong 30,0 (g) bt dc liu chuyn vo mt ti giy lc.
Thm m bng dung dch amoniac 6N (TT), yn trong 40 pht. Chuyn ti giy
lc vo bnh Soxhlet, thm 200 ml cloroform (TT), chit ti khi dch chit lc vi
acid sulfuric cho phn ng m tnh vi thuc th Mayer th dng li. Ly dch chit
c cch thy ti cn, ha tan hon ton cn trong dung dch acid sulfuric 5N (TT).
Lc qua giy lc vo bnh gn. Kim ha dch lc bng amoniac m c (TT) n
pH 10. Chit bng cloroform 4 ln. Gp dch chit cloroform, bc hi cch thy n
cn kh.
+ Dch chit ton phn trong nc thu c bng phng php sc vi dung
mi l nc, bay hi dung mi n dng cao lng 1:1.
+ Dng hn dch: phn tn diazepam, rotundin, cn alcaloid trong CMC
0,5% theo t l thch hp ri pha long bng nc ct t nng mong mun.

17

2.2. Phng php nghin cu

2.2.1. S thit k nghin cu
C ca 2 loi Stephania
sinica v Stephania
dielsiana

Dch chit nc ton phn

Gii lo u, chut
khng chu
stress, EPM

Gii lo u,
chut khng
chu stress,
ETM

Gii lo u,
chut chu
stress, EPM

An thn, chut
khng chu
stress

An thn,
chut chu
stress

Alcaloid ton phn

Gii lo
u, chut
khng
chu
stress,
EPM

An thn,
chut
khng
chu
stress

2.2.2. Ni dung nghin cu

Thit k 4 t th nghim vi cc ni dung sau: nh gi tc dng gii lo u
(m hnh ch thp nng cao, elevated plus maze, EPM) v an thn (tng cng thi
gian ng do thiopental) trn chut bnh thng ca dch chit ton phn vi dung
mi l nc ca 2 loi Stephania cc mc liu khc nhau c i chiu vi tc
dng ca diazepam (t 1). Tc dng gii lo u ca dch chit ton phn sau
c nh gi su hn trn chut chu stress do c lp (m hnh EPM) (t 2) v
trn ri lon lo u lan ta v ri lon hong s trn chut bnh thng (m hnh ch
T nng cao, elavated T maze, ETM) (t 3). Cui cng, nh gi tc dng an thn
(tng cng thi gian ng do thiopental) v gii lo u (m hnh EPM) ca alcaloid
ton phn t 2 loi Stephania c i chiu vi tc dng ca tetrahydropalmatin trn
chut bnh thng (t 4).

18

2.2.3. iu kin tin hnh th nghim

Tt c cc th nghim u c tin hnh trong phng ring bit, yn tnh.
Thi gian tin hnh th nghim trong khong t 8h-17h. Vi m hnh EPM, chut
nht trng c ung thuc trong 5 ngy lin tc v tin hnh cc th nghim vo
ngy th 5, sau khi cho chut ung thuc 60 pht. Vi m hnh ETM, chut cng
trng c ung thuc lin tc trong 21 ngy. Vo ngy th 20, chut c cho tip
xc trc vi m hnh bng cch cho di chuyn t do trn tay m trong 30 pht.
Tin hnh th nghim vo ngy th 21 sau khi cho chut ung thuc 60 pht.
Cng chiu sng trong thi gian lm thc nghim c duy tr bng hai bng
n 60W.
2.2.4. Liu s dng trong nghin cu
Trong th nghim thm d liu (t 1), s dng 4 mc liu ca dch chit
ton phn ca 2 loi Stephania l 30 mg/kg, 100 mg/kg, 300 mg/kg v 1000 mg/kg
tnh theo dc liu kh. Cc th nghim sau (t 2 v t 3) s dng mc liu c
hiu qu c xc nh trong t 1. Liu ca cc alcaloid ton phn (t 4) c
qui i theo liu tng ng vi liu ca dch chit ton phn trong nc c tc
dng. Liu ca cht i chiu diazepam c chn mc 2 mg/kg th trng chut
theo nghin cu ca Dng Phc An v cng s [1], mc liu ny ph hp vi
mc liu ca diazepam dng iu tr lo u trn ngi trng thnh dng ng
ung (4 - 15 mg/ngy) [4], quy i ra liu ca chut nht trng. Mc liu ca chng
dng clomipramin c s dng trong nghin cu l 25 mg/kg, s dng di ngy
(21 ngy) theo nghin cu ca Poltronieri v cng s [44]. Mc liu ny ph hp
theo mc liu quy i t mc liu dng trn ngi iu tr ri lon hong s [4]
sang liu trn chut cng. Mc liu 1 mg/kg ca tetrahydropalmatin c la chn
l mc liu thp nht th hin tc dng gii lo u r rt trong nghin cu ca Leung
WC v cng s [32]. Cc thuc th c phn tn trong CMC 0,5% to hn dch
cho chut ung.

19

2.2.5. M hnh gy stress cho chut bng phng php nui c lp

M hnh gy stress bng phng php nui c lp c trin khai theo m t
ca Ojima v cng s [41]. Chut nht trng khe mnh, ging c chng Swiss
albino 4 tun tui nhn v c nui n nh trong iu kin phng th nghim 5
ngy, sau gy stress c lp cho chut bng cch nui tch ring 1con/lng. L
by n c nui song song 10-15 con/lng trong cng thi gian vi l chut c
nui c lp. Chut c cho n thc n chun, ung nc t do. Cc th nghim
c tin hnh sau 5 tun gy stress.

Hnh 2.3. Hnh nh chut nui c lp gy stress

2.2.6. nh gi tc dng gii lo u trn m hnh EPM
M hnh ch thp nng cao theo m t ca Lister [33].

20

Hnh 2.4. M hnh ch thp nng cao

Dng c th nghim
Dng c bng g gm 2 tay kn kch thc 30 x 5 x 15cm (di x rng x cao),
2 tay m kch thc 30 x 5 x 0,2 cm, 2 tay kn v m t vung gc vi nhau v
t cch nn nh 60 cm to thnh hnh ch thp, cng chiu sng trong thi
gian lm thc nghim c duy tr bng mt bng n 60W.
Phng php tin hnh
Chut c chia ngu nhin thnh cc l v cho ung thuc th lin tc
trong 5 ngy, mi ngy 1 ln vo 8h.
Ngy th 5 sau khi ung thuc 60 pht, chut c t nh nhng vo vng
trung tm, mt hng v cnh tay m, sau c t do di chuyn khm ph trong
5 pht. Chut c coi l trong 1 cnh tay khi c 4 chn u nm trong cnh tay
. Sau khi tin hnh th nghim vi mi chut, dng c th nghim c lau sch
bng khn tm cn trc khi tin hnh vi chut tip theo trnh lu gi mi. Ch
tiu nh gi: ghi nhn thi gian lu li tay m, s ln ra vo cc vng cnh tay
(ng v/hoc m) ca chut.

21

2.2.7. nh gi tc dng gii lo u trn m hnh ETM

M hnh ch T nng cao theo m t ca Graeff v cng s [24].

Hnh 2.5. M hnh ch T nng cao

Dng c th nghim

Hnh 2.6. Dng c ch T nng cao

22

Dng c th nghim bng g c cu to l hnh ch T gm 1 cnh tay kn

kch thc 50 x 12 x 40cm (di x rng x cao) v 2 tay m kch thc 50 x 12 x 1
cm, tay kn c t vung gc vi 2 tay m. B dng c c t cch nn nh
50cm, cng chiu sng trong thi gian lm thc nghim c duy tr bng mt
bng n 60W.
Phng php tin hnh
Pre-test: trc ngy th nghim 1 ngy, chut c phi nhim vi mt
trong hai cnh tay m trong vng 30 pht, mt thanh g chn ngang trc li vo
ca cnh tay kn v mt thanh chn ngang u gn cnh tay m m bo chut
c tch bit vi phn cn li ca dng c. Chut c ung thuc sau khi phi
nhim 30 pht.
T-test: cc th nghim c tin hnh 24h sau pre-test, trc khi tin hnh th
nghim 2 pht, chut c t vo trong mt chic lng sch v kh chun b
trc. Sau chut c t vo cui cnh tay kn, mt hng v trung tm t
do khm ph m hnh. Ghi nhn thi gian k t khi t chut ti khi chut ri tay
kn bng c 4 chn (thi gian tim tng nn, baseline latency), sau ghi nhn li
thi gian ny trong 2 ln t chut vo m hnh lin tip (n trnh 1 v n trnh 2,
avoidance 1 v avoidance 2). Sau hun luyn n trnh, chut c t li vo m
hnh vo phn cui tay m bn phi, mt hng v pha trung tm v ghi nhn thi
gian chut ri tay ny bng c 4 chn (chy trn, escape).Thi gian ti a cho mi
th nghim l 5 pht. Gia cc th nghim, dng c c lau sch bng cn trnh
lu gi mi. Trong khong thi gian 30 giy gia cc th nghim chut c t li
vo lng chun b sn.
2.2.8. nh hng trn thi gian ng do thiopental
Xc nh theo m t ca Trung m v Th Phng [5].
Chut c tim tnh mch ui thiopental liu 40mg/kg. t chut nm
nga, thi gian ng do thiopental c tnh t khi chut bt u ng n khi chut
c phn x lt mnh tr dy.

23

2.3. X l s liu
D liu c lu tr, phn tch bng phn mm SPSS 16.0 v c biu din
di dng gi tr trung bnh sai s chun (X SE). Qua phn tch kim chun
Kolmogorow- Smirnow cho thy mt s d liu khng tun theo phn b chun. V
vy, mu khng tun theo phn b chun v cc mu m bin c tnh cht khng
lin tc c kim nh bng test phi tham s Kruskal Wallis sau l Mann
Whitney U test so snh s khc bit gia cc l. Vi cc mu tun theo phn b
chun, s dng kim nh One way Anova vi hu kim (post hoc) Dunnette T3
so snh s khc bit gia cc l. S khc bit c coi c ngha thng k khi p
< 0,05.

24

CHNG III. KT QU V BN LUN

3.1. nh gi tc dng gii lo u ca dch chit nc ton phn S. dielsiana v
S. sinica
3.1.1. Trn m hnh EPM chut khng chu stress
Thi gian lu ti tay m v s ln ra tay m ca cc l chut th nghim
trong m hnh ch thp nng cao c trnh by trong bng 3.1; s ln vo tay m
v tng s ln ra tay kn, tay m c trnh by trong bng 3.2.
Bng 3.1. Tc dng ca dch chit nc ton phn v diazepam trn thi gian lu
v s ln ra tay m ca chut khng chu stress trn m hnh EPM
Tn l

9
9

Thi gian lu ti
tay m (giy) (a)
8,11 3,85
31,89 12,04

28,11 7,68

19,88 6,28

25,89 4,18

10

34,10 7,15

22,75 9,48

44,50 17,32

53,44 16,52

10

45,60 12,50

NaCl
2
DZP
D30

D100
D300

D1000
S30

S100

S300

S1000

10

pa
p1-2 = 0,020
p1-3 = 0,010
p2-3 = 0,730
p1-4 = 0,093
p2-4 = 0,423
p1-5 = 0,011
p2-5 = 0,489
p1-6 = 0,006
p2-6 = 0,661
p1-7 = 0,139
p2-7 = 0,423
p3-7 = 0,167
p1-8 = 0,015
p2-8 = 0,888
p4-8 = 0,279
p1-9 = 0,006
p2-9 = 0,387
p5-9 = 0,436
p1-10 = 0,006
p2-10 = 0,400
p6-10 = 0,529

S ln ra
tay m (b)
1,11 0,48
3,56 0,75
3,33 0,55
2,50 0,57
3,56 0,41
2,40 0,54
3,38 1,10

3,63 0,89

2,56 0,41

2,70 0,60

pb
p1-2 = 0,008
p1-3 = 0,008
p2-3 = 1,000
p1-4 = 0,044
p2-4 = 0,321
p1-5 = 0,004
p2-5 = 0,546
p1-6 = 0,053
p2-6 = 0,211
p1-7 = 0,093
p2-7 = 0,481
p3-7 = 0,541
p1-8 = 0,021
p2-8 = 0,963
p4-8 = 0,382
p1-9 = 0,031
p2-9 = 0,387
p5-9 = 0,136
p1-10 =0,035
p2-10 =0,278
p6-10 =0,631

*Ghi ch. DZP: Diazepam 2 mg/kg; D30, 100, 300, 1000: S. dielsiana 30, 100, 300, 1000 mg/kg;
S30, 100, 300, 1000: S. sinica 30, 100, 300, 1000 mg/kg.

25

Bng 3.2. Tc dng ca dch chit nc ton phn v diazepam trn s ln vo tay
kn v tng s ln lu ti tay kn, tay m ca chut khng chu stress trn m hnh
EPM

Tn l

S ln vo

p(c) so snh

Tng s ln lu

p(d) so snh

tay kn (c)

vi l NaCl

ti tay kn, tay m

vi l NaCl

NaCl

8,78 1,41

9,89 1,69

DZP

13,78 2,01

0,063

17,33 1,97

0,019

D30

11,11 1,12

0,258

14,44 1,18

0,063

D100

13,38 1,49

0,074

15,88 1,41

0,027

D300

10,33 0,88

0,387

13,89 0,98

0,094

D1000

10

11,00 1,62

0,325

13,40 1,78

0,243

S30

7,88 1,06

0,541

11,25 1,99

0,815

S100

9,63 1,71

0,699

13,25 2,35

0,236

S300

7,89 1,14

0,863

10,44 1,33

0,796

S1000

10

6,80 1,05

0,387

9,50 1,46

0,842

*Ghi ch. DZP: Diazepam 2 mg/kg; D30, 100, 300, 1000: S. dielsiana 30, 100, 300, 1000 mg/kg;
S30,100,300,1000: S. sinica 30, 100, 300, 1000 mg/kg.

26

Nhn xt: Diazepam liu 2mg/kg th hin tc dng gii lo u r rt, lm tng
c ngha thi gian lu ti tay m, tng s ln ra tay m so vi nhm chng ung
nc mui sinh l. Ngoi tr mc liu 100 mg/kg ca S. dielsiana v liu 30 mg/kg
ca S. sinica, tt c cc mc liu cn li u th hin tc r rt trn c s ln di
chuyn vo tay m v thi gian lu li tay ny. cng mt mc liu, tc dng ca
2 loi Stephania khc nhau khng c ngha thng k v tng ng vi tc dng
ca diazepam (2 mg/kg). Ch s s ln vo tay kn ca cc l khng c s khc bit
c ngha so vi l chng NaCl, diazepam v S. dielsiana mc liu 100 mg/kg lm
tng c ngha tng s ln lu ti tay kn, tay m; cc mc liu cn li khng c s
khc bit c ngha so vi l chng ch tiu ny. Mc liu ti thiu c tc dng:
30 mg/kg vi S. dielsiana v 100 mg/kg vi S. sinica c la chn cho cc th
nghim tip theo.
3.1.2. Trn m hnh EPM chut chu stress do c lp
Chut b gy stress do nui c lp trong thi gian 5 tun sau tin hnh cho
ung thuc trong 5 ngy. Kt qu thi gian lu li tay m v s ln vo tay m ca
cc l chut chu stress do c lp v l nui by n trong m hnh ch thp nng
cao c trnh by trong bng 3.3; s ln vo tay kn v tng s ln lu ti tay kn,
tay m c trnh by trong bng 3.4.

27

Bng 3.3. Tc dng ca dch chit nc ton phn v diazepam trn s ln v thi
gian lu ti tay m ca chut chu stress trn m hnh EPM
Thi gian lu ti
Tn l

NaCl - by n
2

NaCl - c lp
3

DZP c lp

S ln ra
pa

tay m (giy) (a)

tay m (b)

19,44 12,91

2,11 1,20

10

p1-2=0,043

p1-2=0,043

10

87,40 41,80

p2-3=0,023

1,20 0,49

p2-3=0,023

p2-4=0,280

10

D30 c lp

3,80 2,89

p2-4=0,280
0,50 0,31

p3-4=0,143

p3-4=0,247

p2-5=0,001

S100 c lp

pb

10,13 2,57

p2-5=0,001
1,25 0,25

p3-5=0,297

p3-5=0,408

Bng 3.4. Tc dng ca dch chit nc ton phn v diazepam trn s ln vo tay
kn v tng s ln lu ti tay kn, tay m ca chut chu stress trn m hnh EPM
Tng s ln
Tn l

S ln vo

p(c) so snh vi

lu ti tay

p(d) so snh

tay kn (c)

NaCl c lp

kn v tay

vi NaCl c lp

m
NaCl-by n

6,33 1,42

0,133

8,44 1,71

0,065

NaCl - c lp

10

4,10 2,00

DZP c lp

10

4,60 1,68

0,971

5,80 1,48

0,165

D30 c lp

10

8,60 1,49

0,052

9,10 1,57

0,052

S100 c lp

10,13 0,72

0,034

11,38 0,75

0,027

4,10 2,00

*Ghi ch. DZP c lp : l chut nui c lp ung diazepam; D30 c lp: l chut nui c lp ung
S. dielsiana 30 mg/kg; S100 c lp: l chut nui c lp ung S. sinica 100 mg/kg

28

Nhn xt: Chut chu stress do c lp biu hin trng thi lo u r rt, th
hin s gim s ln vo tay m v thi gian lu li tay m so vi nhm by n
(p < 0,05). Diazepam (2 mg/kg) vn th hin tc dng r rt gii lo u trn chut
chu stress, lm tng s ln vo tay m cng nh thi gian lu li tay ny (p <
0,05). Trong 2 loi Stephania, ch c S. sinica mc liu th th hin tc dng gii
lo u, lm tng c ngha s ln vo v thi gian lu li tay m so vi l chng
chu stress ung nc mui sinh l. S. dielsiana khng lm thay i c ngha cc
ch tiu ny. S. sinica lm tng c ngha s ln vo tay kn v tng s ln lu ti
tay kn,tay m; diazepam v S. dielsiana khng lm thay i c ngha 2 chi s
ny.
3.1.3. Trn m hnh ETM
p ng n trnh nguy him (s cao khi khm ph tay m) lin quan n
tc dng chng ri lon lo u ton th, th hin bng thi gian tim tng chut
ri tay kn khm ph tay m ti thi im ban u (baseline), ln t chut th
nht (avoidance 1), ln t chut th hai (avoidance 2) v p ng chy trn khi b
phi nhim mi trng nguy him trn tay m lin quan n tc dng chng ri
lon hong s, th hin bng thi gian tim tng chut ri tay m chui vo tay
kn (escape) l chng ung nc mui sinh l v cc l th thuc trong vng 21
ngy c trnh by trong bng 3.5. Liu ca 2 dc liu s dng trong th nghim
c qui i tng ng t liu c tc dng trn chut nht sang chut cng c
xc nh trong th nghim trc trn m hnh EPM chut nht khng chu stress.

29

Bng 3.5. Tc dng ca dch chit nc ton phn v clomipramin trn chut khng
chu stress th hin qua cc ch tiu nh gi trn m hnh ETM

Tn l
NaCl

Clomipramin
25 mg/kg

Avoidance 1

Avoidance 2

Escape (giy)

(giy)

(giy)

204,22 47,90

286,67 13,33

267,78 32,22

20,67 5,56

86,89 40,47

243,67 37,87

143,22 49,62

160,67 44,49

p = 0,297

p = 0,666

p = 0,161

p = 0,002

139,50 44,44

226,40 39,03

270,40 29,60

123,20 39,50

p = 0,447

p = 0,447

p = 1,000

p = 0,013

112,11 47,14

161,89 39,30

225,44 34,81

34,22 13,39

p = 0,297

p = 0,031

p = 0,340

p = 0,489

S. dielsiana
17.5 mg/kg

Baseline (giy)

10

S. sinica
9
58.3 mg/kg

*Gi tr p so snh vi l nc mui sinh l

Nhn xt: Kt qu cho thy, clomipramin (25 mg/kg) v S. dielsiana (17,5

mg/kg) lm tng c ngha thi gian tim tng ca p ng chy trn (escape) m
khng lm thay i p ng n trnh (avoidance) so vi l chng. Ngc li, S.
sinica c tc dng lm gim r rt thi gian tim tng ca p ng n trnh
(avoidance 1) nhng khng lm thay i p ng chy trn. Clomipramin v dch
chit ton phn 2 loi Stephania u khng lm nh hng n thi gian tim tng
khm ph tay m ti thi im ban u (baseline) so vi l chng.
3.2. nh gi tc dng an thn ca dch chit nc ton phn ca S. dielsiana
v S. sinica
3.2.1. Trn chut khng chu stress
nh hng ca dch chit nc ton phn t 2 loi Stephania cc mc liu
khc nhau v ca diazepam ln thi gian ng do thiopental chut khng chu
stress c trnh by trong hnh 3.1.

30

Thoi gian ngu (phut)

25

**

20
15

**

10

**

** **
** *

5
0
0

30 100 300 1000 30 100 300 1000 mg/kg

NaCl DZP DC S. dielsiana

DC S. sinica

Hnh 3.1: nh hng trn thi gian ng do thiopental ca diazepam (DZP) v dch
chit nc ton phn (DC) ca 2 loi Stephania trn chut bnh thng, * p < 0,05,
**

p < 0,01 so vi nhm chng ung nuc mui sinh l.

Nhn xt: Diazepam (liu 2 mg/kg) c tc dng ko di r rt thi gian ng

do thiopental, lm tng thi gian ny ln 3,44 ln so vi l chng ung nc mui
sinh l (p< 0,001). Dch chit S. dielsiana tt c cc mc liu th u c tc dng
ko di thi gian ny t 1,7 - 2,0 ln so vi l chng trong khi dch chit S. sinica
th hin tc dng t r rt hn (ko di thi gian ng do thiopental t 1,23 - 1,51 ln
so vi chng), c xu hng gim dn tc dng khi liu tng dn. mc liu thp
(30 mg/kg v 100 mg/kg), tc dng ca 2 loi Stephania tng ng nhau trong
khi mc liu cao hn (300 mg/kg v 1000 mg/kg), tc dng ca S. dielsiana th
hin r hn S. sinica (p < 0,05). Tc dng ca 2 loi Stephania tt c cc mc liu
th u km hn diazepam (p < 0,05). Mc liu ti thiu c tc dng an thn ca c
2 loi Stephania: 30 mg/kg v 100 mg/kg c la chn cho cc th nghim tip
theo.
3.2.2. Trn chut chu stress do c lp
Chut b gy stress do nui c lp trong thi gian 5 tun sau tin hnh cho
ung thuc trong 5 ngy. Kt qu thi gian ng do thiopental ca cc l chut chu
stress do c lp ung nc mui sinh l, diazepam, dch chit ton phn ca 2 loi
Stephania v l nui by n c trnh by trong hnh 3.2.

31

Thoi gian ngu (phut)

25

**

20
15
10

*
5
0
0
NaCl

0
NaCl

Nuoi bay dan

2
DZP

30

100

S. dielsiana

30

100

mg/kg

S. sinica

Nuoi co lap

Hnh 3.2: nh hng trn thi gian ng do thiopental ca diazepam (DZP) v dch
chit nc ton phn (DC) ca 2 loi Stephania trn chut chu stress do c lp, * p
< 0,05 so vi l chng nui by n ung nc mui sinh l, ** p < 0,05 so vi
nhm chng nui c lp ung nuc mui sinh l.
Nhn xt: Chut chu stress gy ra bi c lp c biu hin gim thi gian ng
do thiopental (24,5%) so vi nhm by n (p < 0,05). Trn nhm chut chu stress
do c lp, c 2 loi Stephania u khng th hin tc dng tng cng thi gian ng
do thiopental c 2 mc liu th. Ngc li, diazepam vn th hin tc dng r rt,
lm ko di thi gian ng do thiopental gp 2,49 ln so vi nhm chng ung nc
mui sinh l cng iu kin chu stress c lp (p < 0,01).
3.3. nh gi tc dng gii lo u v an thn ca alcaloid ton phn t S.
dielsiana v S. sinica
3.3.1. Chit xut alcaloid t c 2 loi Stephania
Trin khai sc k lp mng alcaloid ton phn 2 loi Stephania v cht
chun trn cng mt bn mng:
Cn alcaloid ton phn v cht chun (tetrahydropalmatin) c ha tan
trong cloroform. H dung mi khai trin CHCl3 : MeOH (9:1) [6]. Hin mu vi
thuc th Dragendorf. Kt qu cc vt alcaloid thu c th hin trn hnh 3.3.

32

Hnh 3.3. Sc k lp mng phun thuc th hin mu c i chiu vi cht chun

tetrahydropalmatin
C : tetrahydropalmatin; S : Stephania sinica ; D : Stephania dielsiana
Nhn xt: C th thy r vt ln nht ca Stephania sinica c Rf trng vi
vt ca cht chun tetrahydropalmatin, trong khi Stephania dielsiana khng quan
st thy vt no c Rf trng vi Rf ca cht chun.
Hiu sut chit alcaloid vi S. sinica v S. dielsiana tng ng l 7,1% v
2,8%.
3.3.2. nh gi tc dng gii lo u ca alcaloid ton phn t S. dielsiana v S.
sinica
Tc dng gii lo u ca alcaloid ton phn t 2 loi S. dielsiana v S. sinica
mc liu tng ng vi liu ca dch chit ton phn c tc dng xc nh trong
th nghim trc c nh gi trn m hnh ch thp nng cao chut khng chu
stress. Kt qu v thi gian lu li tay m v s ln vo tay m ca cc l chut th
nghim trong m hnh ch thp nng cao c trnh by trong bng 3.6; s ln vo
tay kn v tng s ln lu ti tay kn, tay m c trnh by trong bng 3.7.

33

Bng 3.6. Tc dng ca alcaloid ton phn, diazepam v tetrahydropalmatin trn s

ln v thi gian lu ti tay m ca chut khng chu stress trn m hnh EPM
Tn l

11

Thi gian lu ti
tay m (giy) (a)
3,27 1,70

13

41,23 15,47

20,75 3,63

NaCl

p(a)

S ln ra tay
m (b)
0,36 0,20

p(b)

Diazepam
2 mg/kg

Rotundin
1 mg/kg 3
4

S.sinica
7,14 mg/kg

13

37,85 21,31

S.dielsiana
0,84 mg/kg

12

13,75 4 ,08

p1-2 = 0,001
p1-3 = 0,000

3,69 0,75

2,13 0,67

p1-2 = 0,000
p1-3 = 0,004

p2-3 = 0,916

p2-3 = 0,121

p1-4 = 0,001

p1-4 = 0,001

p2-4 = 0,650

2,23 0,56

p2-4 = 0,081

p3-4 = 0,595

p3-4 = 0,804

p1-5 = 0,032

p1-5 = 0,027

p2-5 = 0,123
p3-5 = 0,181

1,58 0,42

p2-5 = 0,026
p3-5 = 0,571

*Liu alcaloid quy i theo liu dch chit ton phn tng ng da vo hiu sut chit alcaloid t
c 2 loi Stephania.

34

Bng 3.7. Tc dng ca alcaloid ton phn, diazepam v tetrahydropalmatin trn s

ln vo tay kn v thi gian lu ti tay kn, tay m ca chut khng chu stress trn
m hnh EPM.

Tn l

S ln vo
tay kn (c)

NaCl

p(c) so vi

Tng s ln vo

p(d) so vi

nhm chng

tay kn, tay m

nhm chng

NaCl

(d)

NaCl

11

7,73 0,91

8,09 0,95

13

8,46 1,22

0,459

12,15 1,53

0,072

4,38 1,56

0,026

6,50 2,15

0,206

13

9,54 1,12

0,134

11,77 1,55

0,082

12

9,00 0,59

0,169

10,58 0,69

0,051

Diazepam
2mg/kg
Rotundin
1mg/kg
S.sinica 7,14
mg/kg
S.dielsiana
0,84 mg/kg
*Liu alcaloid quy i theo liu dch chit ton phn (tng ng 2 mc liu 20 mg/kg v
100 mg/kg) da vo hiu sut chit alcaloid t c 2 loi Stephania.

Nhn xt: diazepam (2 mg/kg) v tetrahydropalmatin (1 mg/kg) th hin tc

dng gii lo u r rt thng qua kh nng tng c ngha s ln vo tay m v thi
gian lu li tay ny so vi l chng ung nc mui sinh l. mc liu tng
ng vi liu 30 mg/kg (S. dielsiana) v 100 mg/kg vi (S. sinica) tnh theo dch
chit ton phn, cn alcaloid ton phn cng lm tng r rt c hai ch tiu quan st
(p < 0,05). Tc dng ca cn alcaloid ton phn v tetrahydropalmatin mc liu
th c xu hng yu hn tc dng ca diazepam nhng s khc bit cha t mc
c ngha thng k tr tc dng ca S. dielsiana trn ch tiu s ln vo tay m.
Tetrahydropalmatin khng lm thay i tng s ln vo tay kn, tay m so vi nhm

35

chng NaCl. Diazepam v 2 loi dc liu khng lm thay i c ngha s ln

vo tay kn v tng s ln vo tay kn vo tay m so vi nhm chng NaCl.
3.3.3.nh gi tc dng an thn ca alcaloid ton phn t 2 loi Stephania
Tc dng an thn ca alcaloid ton phn t 2 loi S. dielsiana v S. sinica
mc liu tng ng vi liu ca dch chit ton phn c tc dng xc nh trong
th nghim trc c nh gi trn nh hng ti thi gian ng do thiopental
chut khng chu stress. Kt qu c trnh by trong hnh 3.4.
20
Thoi gian ngu (phut)

**
**

15

**

10

**

**

0
0

NaCl

DZP

THP

0,84

2,80

S. dielsiana

2,14

7,14

mg/kg

S. sinica

Hnh 3.4: nh hng trn thi gian ng do thiopental ca diazepam (DZP),

tetrahydropalmatin (THP) v alcaloid ton phn ca 2 loi Stephania trn chut
bnh thng. * p < 0,05, ** p < 0,01 so vi nhm chng ung nuc mui sinh l.
Nhn xt: mc liu tng ng vi liu 30 mg/kg v 100 mg/kg tnh theo
dch chit ton phn, cn alcaloid ca c 2 loi Stephania u lm tng c ngha
thi gian ng do thiopental (1,31 - 1,87 ln so vi l ung nc mui sinh l). Tc
dng ny km hn c ngha so vi tc dng ca tetrahydropalmatin ( c 2 mc
liu th ca 2 loi S. dielsiana v S. sinica) v ca diazepam ( mc liu thp ca
cn alcaloid t S. sinica). Tc dng ca S. dielsiana mnh hn so vi S. sinica
mc liu thp (p < 0,05). Trong khi mc liu cao, tc dng ca 2 loi
Stephania tng ng nhau.

36

3.4. BN LUN
Gii lo u l tc dng dc l thn kinh quan trng c nhiu nghin cu
tp trung tip cn nhm tm ngun dc liu c hiu qu, t tc dng ph thay th
hoc b sung cho cc thuc ha dc ang c s dng trong iu tr [57]. Nhng
kt qu nghin cu v ha hc v dc l tm thn ca cc loi trong chi Stephania
Lour. ni chung, 2 loi S. sinica v S. dielsiana mi c tp trung nghin cu
Vit nam ni ring l c s cho vic nh gi tc dng gii lo u v an thn thc
nghim c thc hin trong nghin cu ny.
V tc dng gii lo u, kt qu t 3 m hnh dc l thc nghim s dng
trong nghin cu cho thy dch chit ton phn ca S. sinica v S. dielsiana u th
hin tc dng gii lo u. Tuy nhin c s khc bit gia mc v c im gii lo
u gia 2 dc liu ny.
Trn m hnh ch thp nng cao (EPM), m hnh kinh in c s dng
ph bin sng lc tc dng gii lo u da trn p ng lo u khng iu kin ca
ng vt thc nghim do s cao khi khm ph cnh tay m v xu hng lu li
ni n np an ton (tay kn) trong m cung ch thp [18], dch chit ton phn ca
S. dielsiana v S. sinica u lm tng s ln di chuyn vo tay m v thi gian lu
li tay ny so vi l chng, th hin tc dng gii lo u tng t thuc i chiu
diazepam (bng 3.1). Kh nng tng thi gian lu li tay m c th lin quan n
tc dng an thn qu mc lm gim hot ng t nhin ca chut c loi tr do
s ln khm ph tay kn v tng s ln khm ph tay kn v tay m ca m cung
(bng 3.2; bng 3.4) cc nhm th thuc trn chut bnh thng v chut chu
stress do c lp u tng hoc gim khng c ngha so vi nhm chng dng
nc mui sinh l [22], [45]. Trong khong liu la chn th trn m hnh (t 30
mg/kg n 1000 mg/kg), tc dng gii lo u t ph thuc vo liu, ng thi khng
c s khc bit v mc tc dng gia 2 loi cng mt mc liu s dng.
La chn liu ti thiu c tc dng nh gi tip trn m hnh EPM
chut chu stress do c lp, kt qu cho thy ch c S. sinica th hin tc dng gii

37

lo u trn cc ch tiu nh gi tng t nh diazepam (bng 3.3). Chut chu stress

do c lp th hin trng thi lo u r rt hn nhm chut nui by n, th hin r
rt trn s ln ra tay m v thi gian lu li tay ny [2]. M hnh ny c Korte v
cng s nh gi c nhiu u im hn so vi m hnh kinh in trn chut khng
chu stress cho php nh gi hnh vi tng cng s hi, tng cng tnh trng lo u
c to ra trc trn ng vt thc nghim bi tc nhn gy stress, phn nh
cm xc bt an ca ng vt khi ng trc e da hoc nguy him [30]. S thay
i v hnh vi ca chut chu stress, c gi thuyt lin quan n hot ha h
noradrenergic trung ng v yu t gii phng corticotrophin (CRF) vng di
i [41] hay gim kh nng iu bin ca GABA trong vic m knh Cl-, c th
thay i tc dng gii lo u ca cc thuc [49]. iu ny cho thy, c th phi s
dng mc liu cao hn vi S. dielsiana, nh quan st vi ch phm Sleepcare
trong nghin cu trc y quan st c tc dng gii lo u ca dch chit trn
chut chu stress do c lp [2].
M hnh ch thp nng cao kinh in (EPM) vi ch tiu nh gi chnh da
trn s ln vo v thi gian lu li tay m ca ng vt thc nghim c coi l m
hnh trn ln, khng cho php phn bit cc th lo u khc nhau [54]. V vy,
Graeff v cng s pht trin m hnh ch T nng cao (ETM), ci tin da trn
m hnh EPM kinh in phn bit p ng chy trn (escape) khi ng vt thc
nghim c tip xc vi tay m (lin quan n ri lon hong s) v p ng n
trnh c ch (inhibitory avoidance) sau ghi nh phi nhim vi vng nguy him
(lin quan n ri lon lo u ton th) p dng nh gi tc dng gii lo u ca
thuc. Theo , s hi c iu kin (conditioned fear) hay p ng n trnh c ch
c tnh l thi gian chut ri tay kn trong 3 th nghim lin tip (avoidance
1,2,3), cn s hi khng iu kin (unconditioned fear) l khong thi gian chut
trn thot khi tay m hay p ng chy trn. Theo Deakin v Graeff, s hi c
iu kin c lin quan n ri lon lo u lan ta (Generalized Anxiety DisorderGAD) ngi, trong khi s hi khng iu lin lin quan n ri lon hong s
Panic Disorder (PD) [20]. Vic cho ng vt thc nghim tip xc vi m hnh

38

(trong vng 30 pht) 24 h trc khi tin hnh th nghim gip gim thi gian chy
trn do tng nhy pht hin tc dng chng hong s ca cc thuc c
thc hin trong nghin cu [50]. Trn m hnh ny, dch chit ton phn ca S.
dielsiana lm tng c ngha thi gian tim tng ca p ng chy trn (escape)
khi chut c t tay m, th hin tc dng chng ri lon hong s tng t
thuc i chng chng trm cm 3 vng clomipramin [44], m khng lm thay i
p ng n trnh (avoidance) so vi l chng (bng 3.5). Ngc li, dch chit ton
phn ca S. sinica li c tc dng lm gim r rt thi gian tim tng ca p ng
n trnh (avoidance 1) nhng khng lm thay i p ng chy trn, th hin tc
dng chng ri lon lo u lan ta tng t cc dn cht benzodiazepin v buspiron
[24]. Nh vy, c th gi thuyt v s khc bit v thnh phn c hot tnh v c
ch tc dng gii lo u gia 2 loi S. sinica v S. dielsiana.
Thnh phn hot cht chnh to nn tc dng gii lo u ca 2 loi Stephania
l alcaloid. Alcaloid ton phn ca ca c hai dc liu vi mc liu tng ng
vi liu ti thiu c tc dng ca dch chit ton phn u lm tng c ngha s
ln vo tay m v thi gian lu li tay m tng t diazepam v tetrahydropalmatin
(bng 3.6) trong khi khng lm thay i c ngha s ln vo tay kn v tng s ln
vo tay kn, tay m so vi nhm chng NaCl (bng 3.7). Sinomenin v
tetrahydropalmatin, 2 alcaloid phn lp c t c S. sinica v loi S. dielsiana [6],
[16] u c chng minh c tc dng gii lo u trn cc m hnh thc nghim
[19], [32]. liu thp (0,5 - 10 mg/kg), THP c tc dng gii lo u r rt trn m
hnh ch thp nng cao, trong tc dng liu ti thiu c tc dng l 1 mg/kg
c chng ti la chn s dng trong nghin cu ny. mc liu cao hn (50
mg/kg), tc dng gii lo u ca THP gim r rt, thay th bng tc dng an thn v
gin c. Tc dng gii lo u ca tetrahydropalmatin b i khng bi flumazenil,
mt cht cnh tranh v tr gn ca benzodiazepin trn receptor GABAA, chng t c
ch gii lo u ca THP thng qua, t nht mt phn, tng tc vi v tr gn ca
benzodiazepin trn receptor GABAA [26], [32]. Tuy nhin, d liu hin ti cha gii
thch c s khc bit v mc tc dng v c tnh gii lo u ca 2 loi

39

Stephania. S khc bit c th mt phn lin quan n hm lng

tetrahydropalmatin trong hai loi Stephania, nhng cng khng loi tr vai tr ca
cc alcaloid khc vi c ch tc dng khng thng qua trung gian GABAA nh
tetrahydropalmatin [58] gii thch tc dng trn ri lon hong s ca S.
dielsiana tng t cc thuc tc dng thng qua h serotoninergic [43].
V tc dng an thn, kt qu nghin cu cho thy dch chit ton phn ca
S. sinica v S. dielsiana u c tc dng an thn th hin trn kh nng ko di thi
gian ng do thiopental, ng thi alcaloid l thnh phn chnh to tc dng an thn
ca 2 dc liu mc liu th hin tc dng gii lo u. Tuy nhin, trn chut chu
stress do c lp, dch chit ca c 2 loi Stephania u khng th hin tc dng an
thn lm o ngc tc dng rt ngn thi gian ng do thiopental gy ra bi stress
mc liu th hin r tc dng an thn trn chut bnh thng.
Ko di thi gian ng do thuc ng barbituric l th nghim kinh in thm
d tc dng an thn ca thuc [36]. Kt qu trong nghin cu ny khng nh tc
dng an thn ca dch chit ton phn, ca alcaloid ton phn v ca
tetrahydropalmatin (hnh 3.1; hnh 3.2 v hnh 3.4), nhng mc liu thp hn rt
nhiu so vi cc nghin cu trc y cho thy tm quan trng ca vic la chn
liu c tc dng. Nguyn Quc Huy v cng s ghi nhn tc dng ko di thi gian
ng do thiopental gp 3 ln v 8 ln tng ng khi dng liu 1,25 g/kg v 2,5 g/kg
dch chit S. dielsiana (LD50 = 22,2 g/kg) [10]. Nguyn Lan Thy Ty v cng s
quan st thy Stilux-60 (L-tetrahydropalmatin) th hin tc dng ko di thi gian
ng do thiopental gp khong 7 ln khi dng liu 48 mg/kg trn chut nht trng
[15].
Tng t tc dng gii lo u, tc dng an thn ca 2 dc liu ch yu c
gii thch do hot tnh ca alcaloid trong c tetrahydropalmatin. Tuy nhin, tc
dng ny, c bit vi S. dielsiana cn c th c vai tr ca cc alcaloid khc do
hm lng rt thp tetrahydropalmatin loi ny. Tc dng an thn ca
tetrahydropalmatin c chng minh c lin quan n kh nng hot ha

40

receptor GABAA [26]. Trn chut chu stress do c lp, khc vi diazepam, c 2 loi
Stephania u khng th hin tc dng an thn thng qua kh nng ko di thi gian
ng do thiopental 2 mc liu 30 mg/kg v 100 mg/kg (hnh 3.2). Chut chu stress
do c lp trong nghin cu ny vi nhng biu hin thay i v hnh vi v tm l:
tng hot ng vn ng t nhin, tng tm l s hi vi nhng khu vc cao v
c chiu sng trong m hnh ch thp nng cao, c thi gian ng do thiopental
ngn hn so vi nhm chut nui by n [2]. Kt qu t nghin cu Kurmar v
Kulkarni cng cho thy chut chu stress do c lp c biu hin ko di thi gian
tim tng v rt ngn thi gian ng gy ra bi pentobarbital, tng hot ng vn
ng (di chuyn, ng ln) ca chut so vi chut nui by n [31]. S thay i
ny cng c th lm thay i p ng ca ng vt thc nghim vi cc thuc an
thn [46]. C kh nng c ch hot ha h noradrenergic trung ng v yu t gii
phng corticotrophin (CRF) vng di i km theo s gim kh nng iu bin
ca GABA trong vic m knh Cl- lin quan n cht ch vn ngc ni sinh ca
receptor benzodiazepine chut chu stress do c lp khng b o ngc bi 2
loi Stephania [41],[42].
Nh vy, kt qu nghin cu trong ti ny chng minh c tc dng
gii lo u v an thn ca 2 loi S. sinica, S. dielsiana cng nh alcaloid ton phn v
tetrahydropalmatin trn mt s m hnh thc nghim. S khc bit v mc v
c im tc dng gii lo u gia S. sinica v S. dielsiana cn c nghin cu su
hn xc nh vai tr v c ch tc dng gii lo u ca cc alcaloid c hot tnh
gii lo u ca 2 loi ny.

41

KT LUN V XUT
KT LUN
T kt qu thu c trn cc m hnh dc l thc nghim c thc hin trong
ti chng ti rt ra mt s kt lun sau:
1. Dch chit ton phn trong nc ca S. sinica v S. dielsiana cc mc liu
t 30 mg/kg n 1000 mg/kg th trng chut th hin tc dng gii lo u trn
m hnh ch thp nng cao chut bnh thng tng t diazepam, tuy
nhin ch S. sinica c tc dng gii lo u trn chut chu stress do c lp.
Dch chit ton phn ca S. sinica mc liu tng ng 100 mg/kg trn
chut nht c tc dng chng ri lon lo u ton th trong khi S. dielsiana
mc liu 30 mg/kg c tc dng chng ri lon hong s tng t
clomipramin trn m hnh ch T nng cao.
2. Dch chit nc ton phn ca S. sinica v S. dielsiana cc mc liu c tc
dng gii lo u (30- 1000 mg/kg th trng chut) th hin tc dng an thn
trn m hnh ko di thi gian ng do thiopental chut bnh thng nhng
khng c tc dng trn chut chu stress do c lp.
3. Alcaloid ton phn t c hai loi ( mc liu tng ng vi mc liu 30
mg/kg tnh theo dc liu i vi S. dielsiana v 100 mg/kg tnh theo dc
liu i vi S. sinica) u c tc dng gii lo u tng t diazepam v
tetrahydropalmatin trn m hnh ch thp nng cao chut bnh thng.
cc mc liu tng ng vi mc liu 30 mg/kg v 100 mg/kg tnh theo dc
liu, alcaloid ton phn ca c 2 loi u th hin tc dng an thn.
XUT
1. Nghin cu chit xut, phn lp cc alcaloid c hot tnh an thn, gii lo u
khc ngoi tetrahydropalmatin.
2. Nghin cu c ch dc l phn t v vai tr ca cc alcaloid trong tc dng
gii lo u ca 2 loi Stephania.

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