Nghin cu mt s tc dng dc l ca dch chit cy hon ngc

ti sinh vin NCKH Chuyn ngnh: Ha sinh Thc hin: Sinh vin Phng Th Hng Lng Hng dn: PGS.TS. Lng Th Hng Vn

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t vn
Gc t do (GTD) l mt trong nhng nguyn nhn gy ra s lo ha v rt nhiu cc loi bnh nh: ung th, vim khp, tim mch, i tho ng, nhim c Cc GTD c th c loi b nh cc cht ho hc c bit c gi chung l cc cht chng oxy ho (COXH), chng c nhiu trong cc loi rau, c, qu v mt s cy thuc. Cy HN vn c dn gian s dng nh mt cy thuc c kh nng cha rt nhiu bnh khc nhau. Nhng mt s tc dng thc ca chng nh chng oxy ha, c ch mt s vi khun cn cha c chng minh mt cch y .

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Mc tiu nghin cu
1.

Xc nh hm lng mt s cht c tc dng chng oxy ha trong cy Hon ngc

Xc nh kh nng khng mt s vi khun c hi ca dch chit t l v r cy Hon ngc .

2.

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Tng quan ti liu

1.

2.
3. 4. 5.

Gii thiu chung v cy Hon Ngc Gc t do v nguyn nhn hnh thnh gc t do Mt s hng gii quyt gc t do i cng v cc cht chng oxy ho c im ca mt s cht chng oxy ho thng c dng trong dc phm.

I TNG NGHIN CU

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i tng nghin cu Mt s b phn ca cy HN c nghin cu gm c: + L cy HN

+ R cy HN

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PHNG PHP, VT LIU NGHIN CU

1.

Phng php nghin cu:

S dng phng php thc nghim, phn tch, m t, so snh cc mu c lp v so vi i chng.

2.

Vt liu nghin cu
S dng 2 chng vi khun l Escherichia. coli ( E.coli ) v Bacillus. subtilis (B. Subtilis) th hot tnh khng sinh ca l v r cy hon ngc.

3. Ho cht:

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Gm cc ho cht chnh nh: NaCl, Pepton, petrol ete, bubfer, ethanol, kali hidroxid, Natri ascobat, methanol, - tocopherol, ascobic acid chun gc...

4. Thit b nghin cu:

+ My cc ph + My sc k lng hiu cao nng (HPLC) + T m nui vi khun + Bung nui cy vi khun + Cn phn tch + Ni kh trng t + V mt s thit b my mc khc

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H thng my cc ph

H thng my sc k lng hiu cao nng

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K thut tin hnh

thut thu dch chit t l, r cy HN. K thut xc nh hot tnh khng khun ca dch chit t cy HN. K thut xc nh hm lng vitamin E bng my sc k lng hiu cao nng. K thut xc nh hm lng vitamin C bng my cc ph.

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1. Hm lng mt s cht COXH trong cy HN

Biu hm lng vitamin C trong cc mu dch chit
Hm lng VTM C (mg/kg kh)

45 40 35 30 25 20 15 10 5 0

42.0854

Hm lng

0.8888
L ti L kh

4.6478

0.0717
R ti R kh
Tn mu

Hm lng vitamin C trong cc mu dch chit (mg/kg kh)

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Bi u hm l- ng VTM E trong c c mu d chi ch t Hm l- ng 50 VTM 40 E(mg/ 30 kg kh 20 10 0 L t- i Rt- i Rkh L kh M u 60
53.0729

Hm l- ng

13.3977 0 0

Hm lng vitamin E trong cc mu dch chit (mg/kg kh)

2. Kt qu th tc dng khng khun

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nh chp mt mu th nghim th hot tnh khng khun sau 30 gi nui cy. Mu i chng (gia) Cc mu th dch chit cy HN cc nng khc nhau (xung quanh)

2.1. SoTi liu kh s ti: wWw.SinhHoc.edu.vn cc snh chia nng khng khun ca mu dch chit cy HN gia hai loi vi khun
d(cm)
1.4 1.2 1 0.8 0.6 0.4 0.2 0 0,125 0,25 0,5 Nng (g/ml)

Biu so snh kh nng khng khun ca dch chit l ti cy HN chit bng nc 1.333 ct

Vi khun B. subtilis . Vi khun E. coli

0.867 0.733 0.333 0.533

0.433

Kh nng khng khun ca dch chit l ti hon ngc (chit bng nc ct) trn hai loi vi khun

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Biu so snh kh nng khng khun ca dch chit l ti cy HN chit bng NaCl 0.9 % d (cm)
0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 0.9 0.833
Vi khun B. subtilis Vi khun E. coli

0.6 0.533 0.433 0.5

0,125

0,25

0,5

Nng (g/ml)

Kh nng khng khun ca dch chit l ti HN (chit bng NaCl 0.9%) trn hai loi vi khun

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Biu so snh kh nng khng khun ca dch chit r ti cy HN chit bng nc ct
Vi khun B. subtilis .

d(cm) 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0

0.967

Vi khun E. coli . i

0.7330.7
0.567 0.533

0.767

0,125

0,25

0,5 Nng (g/ml)

Kh nng khng khun ca dch chit r ti (chit bng nc ct) trn hai loi vi khun

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Biu so snh kh nng khng khun ca dch chit r ti cy HN chit bng NaCl 0.9%
d(cm)

1.2 1 0.8 0.6 0.4 0.2 0 0.7 0.533 1 0.733 1.033

Vi khun B. subtilis . Vi khun E. coli .

0.677

0,125

0,25

0,5
Nng (g/ml)

Kh nng khng khun ca dch chit r ti (chit bng NaCl 0.9%) trn hai loi vi khun

Ti liu chia s ti: wWw.SinhHoc.edu.vn 2.2. So snh kh nng khng khun ca cc mu dch chit cy HN th trn B. subtilis
Bi so s nh ho t t kh ng vi khun B.subtilis u nh ca d chi l v rt- i c HN chi bng n- c ct ch t y t d (cm) 0.867 0.733 1.333 0.967 HLT HRT

0.8 0.6 0.4 0.2 0

0.733 0.533

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun B. subtilis ca dch chit l v r ti cy HN chit bng nc ct

Bi so s nh ho t t kh ng vi khun B.subtilis u nh ca d chi l v rt- i c HN chi bng NaCl ch t y t 0.9% NLT 1.033 1 0.9 d (cm) NRT 1
0.8 0.6 0.4 0.2 0

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0.7 0.433 0.5

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun B. subtilis ca dch chit l v r ti cy HN chit bng NaCl 0.9%

Ti liu chia s ti: wWw.SinhHoc.edu.vn 2.3. So snh kh nng khng khun ca cc mu dch chit cy HN th trn E. coli

Bi so s nh ho t t kh ng vi khun E.coli ca u nh d chi l v rt- i c HN chi bng n- c ct ch t y t d (cm)

1

0.8 0.6 0.4 0.2 0

0.7 0.567 0.333 0.433 0.533

0.767

HLT HRT

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun E. coli ca dch chit l v r ti cy HN chit bng nc ct

Ti liu chia s ti: wWw.SinhHoc.edu.vn Bi so s nh ho t t kh ng vi khun E.coli ca u nh d chi l v rt- i c HN chi bng NaCl 0.9% ch t y t
d (cm) 1
0.8 0.6 0.4 0.2 0

0.533 0.533 0.6 67

0.677

0.833 0.733

NLT NRT

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun E. coli ca dch chit l v r ti cy HN chit bng NaCl 0.9%

3.4. So snh liu chia s khng khun ca cc mu dch Ti hot tnh ti: wWw.SinhHoc.edu.vn chit khi chit cc dung mi khc nhau
Bi so s nh ho t t kh ng vi khun B.subtilis ca d u nh ch chi l t- i c HN gia hai dung mi kh c nhau t y d (cm)
1.4 1.2 1 0.8 0.6 0.4 0.2 0

1.333 0.867 0.5 0.9

n- c c t NaCl 0.9%

0.733 0.433

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun B.subtilis ca dch chit l ti cy HN gia hai dung mi khc nhau

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Bi so s nh ho t t kh ng vi khun B.subtilis ca u nh d chi rt- i cy HN gia hai dung mi kh c nhau ch t d (cm) 1.033 n- c ct
1.2 0.8 0.6 0.4 0.2 0

0.967

NaCl 0.9%

0.7 0.533

0.733

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun B.subtilis ca dch chit r ti cy HN gia hai dung mi khc nhau

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Bi so s nh ho t t kh ng vi khun E.coli ca d u nh ch chi l t- i cy HN gia hai dung mi kh c nhau t d (cm) n- c ct

1.2 0.8 0.6 0.4 0.2 0 1

0.833 0.533 0.333 0.6 0.433 0.533

NaCl 0.9%

0.125

0.25

0.5

Nng (g/ml)

Hot tnh khng vi khun E. coli ca dch chit l ti cy HN gia hai dung mi khc nhau

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Bi so s nh ho t t kh ng vi khun E.coli ca d u nh ch chi rt- i cy HN gia hai dung mi kh c nhau t d (cm)

1.2 1 0.8 0.6 0.4 0.2 0

n- c ct
0.7 0.677 0.533 0.767

NaCl 0.9%
0.733

0.576

0.125

0.25

0.5 Nng (g/ml)

Hot tnh khng vi khun E. coli ca dch chit r ti cy HN gia hai dung mi khc nhau

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Nhn xt chung

Cc kt qu nghin cu ca chng ti cho thy: + Cc b phn ca cy HN c tc dng dc l r rng bi n c kh nng c ch cc vi khun gy bnh v n c cha mt s cht COXH. + Hm lng cc cht COXH trong l kh cao nhng trong r th rt t. + Cng t nghin cu ca chng ti cho thy cy HN ch c tc dng dc l tt khi cn ti. Khi sy kh th tc dng ca n

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1.

V hm lng cc cht COXH trong cy HN:

- Tt c cc mu l v r u cha vitamin C. Trong , mu l c hm lng vitamin C cao hn rt nhiu so vi mu r; mu ti c hm lng cao hn nhiu so vi mu kh (P < 0.001). - Vitamin E ch c trong mu l (c ti v kh) vi hm lng kh cao trong hm lng vitamin E l ti cao hn rt nhiu so vi l kh, cn trong r (c ti v kh) khng tm thy vitamin E (P < 0.001).

2.

V kh nng khng khun ca dch chit cy HN

- Tt c cc mu ti (l v r) ca cy HN chit bng nc ct v NaCl 0,9% u c kh nng khng hai chng vi khun th nghim (B. subtilis v E.coli), cn i vi mu kh (l v r) u khng c kh nng khng hai chng vi khun th nghim ni trn.

Ti liu chia s ti: wWw.SinhHoc.edu.vn - Hot tnh khng B. subtilis ca cc mu dch chit cy HN ti c s khc nhau: + Mu l ti chit bng nc ct c hot tnh cao hn so vi mu l ti chit bng NaCl 0.9% v cao hn mu r ti chit bng nc ct (p<0,05). + Mu r ti chit bng NaCl 0.9% c hot tnh cao hn so vi mu r ti chit bng nc ct v cao hn mu l ti chit bng NaCl 0.9% (P < 0.05).

- Hot tnh khng E. coli ca tt c cc mu ti (l v r) c chit bng nc ct v NaCl 0.9% u nh nhau (P > 0.05). - Cc mu l ti chit bng nc ct v r ti chit bng NaCl 0.9% c hot tnh khng B. subtilis cao hn so vi E. coli (P < 0.05). - Cc mu l ti chit bng NaCl 0.9% v r ti chit bng nc ct c hot tnh khng hai loi vi khun trn l nh nhau (P > 0.05).

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Kin ngh

Nn tip tc th hot tnh khng khun ca cy HN vi cc chng vi khun khc. Tip tc xc nh hm lng mt s thnh phn ho sinh khc c kh nng COXH v khng sinh trong cy HN nh selenium, carotenoit, flavonoit, alkaloit... Th nghim tc dng COXH v khng khun ca cy HN trn ng vt thc nghim