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KHO ST HIU QU H NG HUYT

V CHNG OXY HA CA CAO CHIT CY NHU
(MORINDA CITRIFOLIA L.) CHUT
BNH TIU NG
i Th Xun Trang1, Nguyn Th Mai Phng1, V Th Ngc Dim1 v Quch T Hu1

ABSTRACT
Hyperglycemia increase in oxidative stress results in diabetic complication. This study
evaluated the anti-diabetic potential of ethanolic extracts from root, leaf, green and ripe
fruits of Morinda citrifolia by determining their antioxidant and antihyperglycemic
activities in vitro and in vivo. The antioxidant activity was determined through the
measurement of DPPH radical scavenging activity in vitro. Mice were induced diabetes
by using alloxan monohydrate. The extracts of Morinda citrifolia could suppress
hyperglycemia in diabetic mice by the oral intake. Finally, the antioxidant effect of these
plant extracts in the kidneys of experimental diabetic mice was observed through the total
antioxidant status (TAS) in kidney of mice. The results showed that oral administration of
ethanolic extracts (200 mg/kg 2 times/ day) could make lower the glucose level of blood
as well as the commercial antidiabetic drug (glucofast), and reduced oxidative stressmediated damage in diabetic kidneys. The results also proved that Morinda citrifolia
ethanolic extracts improved the DPPH radical scavenging activity in vitro.
Keywords: Diabetic, DPPH, TAS, antihyperglycemic, Morinda citrifolia
Title: Studies on hypoglycemic and antioxidant activities of ethnolic extracts from
Morinda citrifolia L. in diabetic mice

TM TT
Tng ng huyt lm gia tng s to thnh stress oxy ha l nguyn nhn ca nhng
bin chng phc tp bnh tiu ng (BT). Nghin cu ny kho st hiu qu iu tr
BT ca cao ethanol r, l, tri xanh v tri chn cy Nhu thng qua kh nng chng
oxy ha v h ng huyt in vivo v vi vitro. Hot ng chng oxy ha c xc nh
thng qua kh nng lm sch gc t do DPPH in vitro. Chut c gy bnh tiu ng
bng dung dch alloxan monohydrate (AM). Sau , chut BT c iu tr bnh vi
cao ethanol cc b phn cy Nhu. Thn ca nhm chut iu tr BT c kho st
hiu qu chng oxy ha thng qua kh nng chng oxy ha tng s (TAS). Kt qu cho
thy, sau khi chut BT ung cao chit cy Nhu vi liu lng 200 mg/kg 2 ln/ ngy
c ng huyt tng ng nhm chut BT c iu tr vi thuc tr tiu ng
glucofast v gim thiu stress oxy ha trong thn. Kt qu cng chng minh rng dch
chit ethanol cy Nhu ci thin kh nng lm sch gc t do DPPH in vitro.
T kha: Bnh tiu ng, DPPH, TAS, s tng ng huyt, cy Nhu

1 GII THIU
Hin nay BT ang pht trin rt nhanh v l nguyn nhn ca nhiu bin chng
phc tp (Bhat et al., 2008). S pht trin ca BT do nhiu nguyn nhn trong
nguyn nhn chnh l stress oxy ha. Stress oxy ha dn n s to thnh cc gc
1

Khoa Khoa hc T nhin, Trng i hc Cn Th

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t do l yu t chnh ca s pht trin bnh v nhng bin chng phc tp ca

BT (Tripathi and Chandra, 2009). Ngn chn s tin trin ca BT bng cch b
sung cc cht chng oxy ha t nhin hin din trong thc vt do cc cht chng
oxy ha ny c kh nng lm sch cc gc t do c hi cho c th t s stress oxy
ha (Pal et al., 2011).
Cy Nhu (Morinda citrifolia L.) v cc b phn ca cy c cc nh khoa hc
c bit quan tm trong nhng nm gn y. Cy Nhu cha nhiu hp cht chng
oxy ha nh beta- carotene, acid ascorbic, terpenoid, alkaloids, beta sitosterol,
carotene, polyphenol nh flavonoid, flavone glycosides, rutin (Ramamoorthy
and Bono, 2007). Cy Nhu c bit c hiu qu, gi thnh thp li t tc dng
ph nn s l mt trong nhng loi thuc thc vt tr BT trin vng
(Ramamoorthy and Bono, 2007).
2 PHNG TIN V PHNG PHP NGHIN CU
2.1 Phng tin
Thit b c s dng trong nghin cu gm my c quay chn khng Heidolph
(c), my o ng huyt ACCU CHECK Active, my ly tm lnh Mikro
220R (c), my o pH Metler Toledo, my lc 8 v tr Daiki SI001 (Hn Quc),
my o quang ph, my khuy t, my vortex.
Ha cht s dng trong th nghim gm ethanol (China), DPPH (2,2Diphenyl
1picrylhydrazyl) (Wako, Japan), hydrogen peroxide (H2O2) (Merck), EDTA
(Merck), Fe(NH4)2(SO4)2 (Merck), acid acetic, alloxan monohydrate (AM)
(Sigma), Trolox (Denmark), nc mui sinh l, sodium benzoate, TBA
(thiobarbituric acid) (Merck), thuc iu tr bnh tiu ng glucofast, diethyl
ether v mt s ha cht khc.
Vt liu th nghim l chut nht trng do vin Pasteur thnh ph H Ch Minh
cung cp. R, l, tri xanh v tri chn cy Nhu c thu hi tnh Kin Giang.
2.2 Phng php
2.2.1 Trch cao cy Nhu bng dung mi ethanol
Nguyn liu c s dng bao gm l, r, tri xanh v tri chn ca cy Nhu c
thu hi tnh Kin Giang. Cc mu c ra sch bng nc sau c ct nh,
ring, phi kh cho n khi trng lng khng i.
250 g mi loi l, r, tri xanh v chn ca cy Nhu kh c ngm trong ethanol
70% nhit phng. Sau thi gian 7 ngy, phn nc dch ngm mu c lc
v c ui dung mi di p sut km. Cao ethanol t cy Nhu sau khi loi b
dung mi c tr 4C s dng cho th nghim sau.
2.2.2 Kho st hot ng lm sch gc t do DPPH in vitro
Hot ng lm sch gc t do ca cc cao ethanol t cc b phn ca cy Nhu
c thc hin theo quy trnh ca Shirwaikar et al. (2006) c iu chnh nh sau:
Cao ethanol cc b phn ca cy Nhu c pha thnh cc nng l 0,25; 0,5;
0,75; 1; 1,5; 2 v 3 mg/ml trong ethanol. 200 l cao chit mi nng kho st
c thm vo 100 l DPPH 6.10-4M. Hn hp phn ng sau khi 60 pht nhit
phng v trong ti c o hp thu quang ph bc sng 517 nm. Hot
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ng lm sch gc t do DPPH c tnh ton tng ng M Trolox

(Koracevic et al., 2000).
2.2.3 Kho st kh nng h ng huyt ca cao ethanol cc b phn cy Nhu
Chut 9 tun tui khe mnh c tim dung dch alloxan monohydrate (AM)
nng 130 mg/kg trng lng chut gy BT. Sau khi chut BT n nh 7
ngy, kh nng h ng huyt ca cao ethanol l, r, tri xanh v tri chn cy
Nhu c xc nh bng cch cho chut BT ung thuc iu tr BT glucofast
(150 mg/kg trng lng chut) hoc cao ethanol t cc b phn ca cy Nhu (200
mg/kg trng lng chut). Chut BT ung cao ethanol l, r, tri xanh v tri
chn 0,1 ml2 ln/ngy trong 21 ngy iu tr. ng huyt c xc nh vo lc
8 9 gi sng trc khi chut c cho n. Sau khi o ng huyt 60 pht, chut
c cho n v ung nc bnh thng. Sau 21 ngy c iu tr bnh vi cc
cao chit, chut c cho nhn i qua m, thn c ly ra v x l cho cc th
nghim tip theo.
2.2.4 Kho st s chng oxy ha tng s (Total Antioxidant Status assay (TAS)) in
vivo
Kh nng chng oxy ha ca cao chit cy Nhu c thc hin da trn nguyn
tc sau: Dung dch sodium benzoate kt hp vi Fe(NH4)2(SO4)2 to ra cc O2(superoxide). Nu trong thn c hin din cc cht chng oxy ha th s kt hp
vi cc superoxide. Lng superoxide cn li s kt hp vi TBA thnh lp
TBA RS (thiobarbituric acid reactive substances) c o bc sng 532 nm.
Quy trnh phn ng c thc hin theo cc bc sau: 100 mg thn chut c
tch ra nghin mn vi 200 l dung dch m phosphate lnh nng 100 mM,
pH = 7,4. Sau khi ly tm trong 10 pht 10.000 vng/pht, phn dch ni trn
c s dng cho vic kho st hot tnh chng oxy ha ca cao chit (Bhat et al.,
2008).
TAS c xc nh theo phng php ca Koracevic et al. (2000) c hiu chnh
nh sau: 10 L dch thn sau khi ng nht (kidney homogenate) c pha long
vi 490 L dung dch m sodium phosphate nng 100 mM, pH = 7,4 c
cho vo hn hp gm 0,5 mL dung dch sodium benzoate 10 mM vi 0,2 mL Fe
EDTA (2 mL Fe EDTA c pha t 2 mM dung dch EDTA vi 2 mM dung
dch Fe(NH4)2(SO4)2). Sau 0,2 mL H2O2 10 mM c cho vo hn hp phn
ng, lc u v 37C trong 60 pht. Sau khi , 1 mL acid acetic 20% v TBA
(thiobarbituric acid) 0,8% trong NaOH c thm vo ng nghim. Hn hp phn
ng sau khi 100C trong 30 pht c ngui nhit phng. hp thu
quang ph ca phn ng c o bc sng 532 nm. TAS trong thn c tnh
ton tng ng nng mM Trolox (Erejuwa et al., 2011).
2.2.5 Thng k phn tch s liu
S liu c trnh by bng MEAN SEM. Kt qu c x l thng k theo
phng php ANOVA bng phn mm minitab 16.0.

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3 KT QU V THO LUN
3.1 S an ton ca cao ethanol cc b phn cy Nhu trn chut bnh thng
nh gi s an ton (khng gy c tnh cp) ca cao ethanol cc b phn cy
Nhu, chut nht trng bnh thng c cho ung cao ethanol nng
200 mg/kg trng lng. Kt qu v s thay i ng huyt ca chut c trnh
by trong hnh 1.
Theo kt qu hnh 1 cho thy, ng huyt ca chut ung cc cao chit v nhm
chut i chng thay i khng khc bit c ngha thng k gia cc nhm v
cc ngy ung cao chit trong sut qu trnh th nghim 21 ngy. Nh vy cao
ethanol cc b phn cy Nhu khng nh hng n ng huyt chut bnh
thng. Mt khc, trng lng chut sau 21 ngy ung cao ethanol cc b phn
cy Nhu cng khng khc bit c ngha thng k so vi nhm i chng (kt
qu khng trnh by).
Nng ng huyt (mg/dl)

160
140
120
100

Ngy 1

80

Ngy 7

60

Ngy 14

40

Ngy 21

20
0

i chng

L
Nghim thc

Tri xanh

Tri chn

Hnh 1: Nng ng huyt ca chut khi ung cc cao chit nng 200 mg/kg
trng lng

T tt c kt qu trnh by trn cho thy cao ethanol cc b phn cy Nhu khng

gy c trn chut bnh thng nng 200 mg/kg trng lng trong 21 ngy.
3.2 Kh nng h ng huyt ca cc cao chit t cc b phn cy Nhu trn
chut bnh tiu ng
Nng ng huyt ca chut sau 7 ngy c tim dung dch AM vo khoang
bng c trnh by trong bng 1. Kt qu cho thy nng ng huyt c s
khc bit c ngha thng k (P < 0,05) khi tim AM nng 130 mg/kg trng
lng so vi chut nhm i chng khng tim AM.
Bng 1: Nng ng huyt ca chut trc v sau khi tim alloxan monohydrate

Nghim thc

Trc khi tim AM

Sau khi tim AM

i chng

124,33 4,04

121,33b 1,15

Tim AM

18

133,44 13,95

421,94a 65,90

Ghi ch: cc ch ci theo sau trong cng mt ct khc nhau s khc bit c ngha thng k mc 5%.

Qua 21 ngy iu tr, kt qu cho thy nhm chut bnh thng ung nc ct
ng huyt gn nh n nh trong sut qu trnh th nghim (khng c khc bit
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c ngha thng k P > 0,05). i vi cc nhm chut BT khng c iu tr

xut hin chut cht ri rc sau 2 ngy v tt c chut cht sau 9 ngy th nghim.
Nhm iu tr bng thuc glucofast c nng ng huyt gim cao nht l
67,65%. Trong cc nhm chut iu tr bng cao chit cy Nhu th nhm iu tr
bng cao r c hiu qu cao nht (gim 58,01%), sau l nhm iu tr vi cao
tri chn (gim 51,62%), cao tri xanh (gim 41,43%) v cui cng l cao l (gim
34,21%).
Bng 2: ng huyt ca cc nhm nghim thc trc v sau khi iu tr bnh tiu ng

Nghim thc
Bnh thng
Khng tr bnh
Glucofast
R
L
Tri xanh
Tri chn

ng huyt (mg/dl)
Trc iu tr
Sau iu tr
121,33b 1,15
132,67c 11,85
464,33a 76,81
Cht
477,00a 28,58
154,33c 4,04
186,00bc 16,52
443,00a 53,81
a
250,67a 22,03
381,00 61,61
a
385,33 90,05
225,67ab 45,24
a
381,00 21,28
184,33bc 57,66

T l tng (+),
gim () (%)
+ 9,35
67,65
58,01
34,21
41,43
51,62

Ghi ch: cc ch ci theo sau trong cng mt ct khc nhau s khc bit c ngha thng k mc 5%.

Theo nghin cu ca Phm Th Lan Anh (2011) trn chut BT c gy bnh

bng AM v iu tr vi cao chit r Nhu vi liu lng 400 mg/kg trng lng,
s dng 0,1 ml/2 ln/ ngy trong 20 ngy iu tr ng huyt gim 71,62%. Nh
vy hiu qu h ng huyt ca cao ethanol r Nhu c s dng trong nghin
cu ny thp hn (58,01%) do liu lng cao chit c s dng thp hn (200
mg/kg trng lng chut).
Nhiu cao chit thc vt cng c chng minh c kh nng h ng huyt trn
chut bnh tiu ng. Trong kh nng h ng huyt ca mp ng
(Momordica charantia) (gim 51,2%), c ca ri (Trigonella foenum graecum) (gim
55,2%) (Tripathi and Chandra, 2009) tng ng vi cao r Nhu (gim 58,01%)
v cao tri chn (gim 51,2%) trong nghin cu ny. Tuy nhin kh nng h ng
huyt ca cao chit t cc b phn cy Nhu thp hn so vi nho (Vitis vinifera
variety Burgund mare) (gim 65,33%) (Chis et al., 2009) v cy l da (Pandanus
amaryllifolius R.) (gim 73,2% khi s dng nng 400 mg/kg trng lng chut)
(Phm Th Lan Anh, 2011).
3.3 Kho st hot tnh chng oxy ha ca cao chit cy Nhu
3.3.1 Kho st kh nng lm sch gc t do DPPH (2,2Diphenyl1-picrylhydrazyl)
ca cao chit t cc b phn cy Nhu in vitro
Xc nh kh nng chng oxy ha bng phng php DPPH l phng php n
gin, nhanh chng v t tn km. Trong nhng nm gn y phng php ny
cng c s dng nh lng cht chng oxy ha trong cc h thng sinh hc
phc tp (Prakash et al., 2000). DPPH l hp cht c mu tm c pht hin
bc sng 517 nm. Khi cc in t l ca cc gc t do DPPH kt hp vi hydro
t cht chng oxy ha th s hnh thnh nn DPPHH lc ny mu sc chuyn t
mu tm sang mu vng. S bin i mu ny tng ng vi lng electron kt
hp vi DPPH (Prakash et al., 2000). Do , kh nng lm sch gc t do ca mt
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cht cng cao th s hp thu quang ph c o 517 nm ca phn ng DPPH c

gi tr cng gim v ngc li.
Trolox (Vitamin E) l cht chng oxy ha chun thng c s dng so snh
kh nng lm sch gc t do ca cc cht cn c kho st. ng chun DPPH
c tnh theo M Trolox theo phng trnh ng chun l y = 0,0096x +
1,6226 (R2 = 0,9918).
Kh nng lm sch gc t do cc nng ca cao ethanol cc b phn cy Nhu
tnh tng ng theo M Trolox da vo phng trnh ng chun, kt qu
c trnh by trong bng 3.
Bng 3: Nng ca cao chit r, l, tri xanh v tri chn cy Nhu tnh tng ng
Trolox

Nng cao
chit (mg/ml)

Gi tr nng cao theo chun Trolox (M)

0,25
0,5
0,75

Cao r
21,38 4,25
34,14 3,71
50,19 3,67

Cao l
36,77 6,38
61,01 6,54
81,67 8,28

Cao tri xanh

8,68 1,99
28,22 0,93
65,49 0,55

Cao tri chn

59,5 4,38
122,39 2,8
150,8 0,43

1,0
1,5

74,31 2,61
93,59 2,93

100,5 7,21
130,44 4,87

70,41 1,97
100,66 0,95

155,11 0,6
155,67 0,38

2,0

105,05 1,36

145,66 5,62

137,73 0,94

160,35 0,49

3,0

133,64 1,56

153,96 4,98

157,55 0,6

162,78 0,5

0,9172 0,007 1,0252 0,001

0,2369 0,002

EC50 (mg/ml)

1,2531 0,003

Kt qu cho thy kh nng lm sch gc t do t l thun vi nng ca cao

chit, nng ca cao chit cng cao th kh nng lm sch gc t do cng ln v
ngc li. Kh nng lm sch 50% cc gc t do EC50 (Effective concentration of
50%) c tnh ton da vo th v kt qu c trnh by trong Bng 2. Trong
cao chit tri chn c kh nng lm sch gc t do cao nht (EC50= 0,2369
0,002) sau l cao l (EC50= 0,9172 0,007), cao tri xanh (EC50= 1,0252
0,001) v cui cng l cao r (EC50= 1,253 0,003).
Phn trm hp th DPPH ca cao ethanol cc b phn cy Nhu c th hin
hnh 2 cho thy khi nng cao chit tng th hp th DPPH cng tng.
Trong , phn trm hp th DPPH ca cao tri chn l ln nht nng 0,25
mg/ml (36,57%), thp nht l cao tri xanh (7,14%). nng 3 mg/ml cao chit,
hp th DPPH ca cao tri chn l 96,39%, thp nht l cao r 79,51%.

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120
100
80

cao r
cao l

60

cao tri xanh

40

cao tri chn

20
0
0,25

0,5

0,75

1,5

Nng cao chit (mg/ml)

Hnh 2: Kh nng lm sch gc t do ca cao ethanol cc b phn cy Nhu (%)

EC50 ca cao l trong phn ng DPPH theo nghin cu ca Thani et al.

(2010) (0,31 mg/ml) thp hn so vi cao l s dng trong th nghim ny
(0,91716 mg/ml) v c th cao l c s dng trong th nghim ca Thani et al.
(2010) l cao ethanol 95%. Dch chit ca l Nhu thu c t nhng k thut tch
chit khc nhau cng c s dng kho st kh nng lm sch gc t do
DPPH theo nghin cu ca Pak-Dek et al. (2011) cng ph hp vi gi tr EC50
ca cao l trong nghin cu ny.
3.3.2 Kho st kh nng chng oxy ha tng s (Total Antioxidant Status (TAS)
assay) in vivo
Thn l mt trong nhng c quan ca c th b nh hng bi stress oxy ha dn
n nhng bin chng phc tp, nn s loi b cc gc t do c hi bi nhng cht
c kh nng chng oxy ha t nhin nhm ngn chn nhng bin chng phc tp
ca BT cng l mt trong nhng mc tiu c la chn hng ti kim sot
BT hiu qu (Pah et al., 2011). Kh nng chng oxy ha ca cao chit cy Nhu
c kho st thn ca chut BT theo phng php TAS. Kt qu v kh nng
chng oxy ha ca cao ethanol t cc b phn cy Nhu c tnh tng ng
theo mM Trolox da vo ng chun v phng trnh ng chun y = 0,3923x
+ 1,1822 (R2 = 0,9902), kt qu c trnh by trong bng 4.
Kt qu bng 4 cho thy nhm chut BT khng c iu tr c kh nng
chng oxy ha thp nht (0,5240 0,0123 mM Trolox) c khc bit c ngha
thng k so vi cc nghim thc cn li (P < 0,05). Kh nng chng oxy ha c
ci thin cc nhm c iu tr bnh vi glucofast v cc cao chit. Trong
nhm chut iu tr bng glucofast c kh nng chng oxy ha tng ng vi
nhm chut bnh thng ung nc ct (khng c khc bit c ngha thng k
P > 0,05). Kh nng chng oxy ha ca nhm chut BT iu tr vi cao r v cao
tri chn tng ng nhau, nhm chut BT iu tr vi cao l tng ng vi
nhm iu tr vi cao tri xanh. Tuy nhin, cc nhm iu tr vi cao ethanol t
cc b phn cy Nhu u c khc bit c ngha thng k so vi nhm chut
bnh thng ung nc ct (P < 0,05).

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Bng 4: Hiu qu chng oxy ha ca cao chit t cc b phn cy Nhu c kho st

thn chut bnh tiu ng

Nghim thc

Hiu qu chng oxy ha c

tnh theo mM Trolox
1,1218a 0,0215
0,5240d 0,0123
1,0963a 0,0015
1,0536b 0,0051
1,0120c 0,0256
1,0251bc 0,0259
1,0522bc 0,0497

Bnh thng
Bnh khng tr
Glucofast
R
L
Tri xanh
Tri chn

Hiu qu chng oxy ha

c tnh theo t l %
88,33
40,48
86,29
82,88
79,55
80,59
82,76

Ghi ch: cc ch ci theo sau trong cng mt ct khc nhau s khc bit c ngha thng k mc 5%.

Lng gc t do kt hp vi cht chng oxy ha c trong thn ca chut tng

ng cc nghim thc c th hin trong bng 4 v hnh 3. Kt qu cho thy
phn trm lng gc t do kt hp vi cht chng oxy ha cao nht nhm chut
bnh thng ung nc ct (88,33 %) v thp nht nhm chut BT khng c
iu tr (40,48 %). i vi cc nhm chut BT c iu tr vi glucofast
(86,29%) v gim dn cc nhm chut BT iu tr vi cao chit cy Nhu ln
lt l r (82,88%), tri chn (82,76%) tri xanh (80,59%) v l (79,55%).
Gc t do kt hp vi cao chit (%)

100
80
60
40
20
0
Bnh Khng tr Glucofast
thng
bnh

Tri xanh Tri chn

Nghim thc

Hnh 3: Phn trm lng gc t do kt hp vi cht chng oxy ha trong thn

Theo nghin cu ca Erejuwa et al. (2011) chut BT c iu tr vi thuc iu

tr BT nh metformin hoc glibenclamide kt hp vi mt ong ci thin c
tnh trng stress oxy ha thn hiu qu hn so vi ch s dng thuc iu tr
metformin hoc glibebclamide.
T kt qu nghin cu t c cho thy cao chit t cc b phn ca cy Nhu c
hiu qu n nh ng huyt chut BT cng nh loi b c cc tc nhn
gy ra stress oxy ha thn ca chut BT. iu ny cho thy hiu qu ca vic
s dng cy Nhu nh liu php h tr trong iu tr BT. y chnh l thng tin
khoa hc hu ch cho cc bnh nhn BT la chn cy Nhu h tr iu tr
bnh. Nghin cu ny cng gp phn b sung c s khoa hc cho y hc c truyn
nhng d liu lin quan n vic ng dng cy Nhu trong iu tr bnh.

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4 KT LUN V KIN NGH

4.1 Kt lun
Cao ethanol ca cy Nhu c tc dng h ng huyt trn chut BT sau 21
ngy iu tr theo th t r (58,01%), tri chn (51,62%), tri xanh (41,43% v
l (34,21%) .
Hiu qu loi b gc t do ca cao chit Nhu theo th t tri chn (EC50 =
0,237 0,002), l (EC50 = 0,917 0,006), tri xanh (EC50 = 1,025 0,001) v
r (EC50 = 1,531 0,003).
Phn trm lng gc t do to ra thn ca chut bnh tiu ng kt hp vi
cc cht chng oxy ha trong cao chit cy Nhu theo th t ln lt tri chn
(82,76%), r (82,88%) tri xanh (80,59) v l (79,55%). Hiu qu loi b gc
t do ca cc cao chit Nhu tng ng vi glucofast (86,29%) v gn bng
vi chut khe mnh (88,33%).
4.2 Kin ngh
Kho st cc c ch h ng huyt ca cao chit cy Nhu trn chut bnh
tiu ng.
Kho st c ch chng oxy ha ca cao chit cy Nhu cc c quan khc
nhau ca chut bnh tiu ng.
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