NgUen ciht YDHCT Viet Nam So 47 - 2015
^BQC TINH BAN TRlTOfNG DIEN VA TAC DUNG CAITHIEN TRI N H 6 CtJA HOAX HUYET AN NAG TREN THlTC NGHIEM Sub-chronic toxicity and memory-enhancing activity of Hoat Huyet An Nao in
experimental animals
* Tran Minh Hilu \ ** Pham Thi Van Anh \
*** TrSn Quoc Binh \ **** TrSn Th6 Linh '
Benh vien Yhoc c6 truyen Trung uang, ^ Tncffng Dai hoc Y Ha Noi
Nghien cieu duac thi^c Men nhSm muc tieu: (}) Xdc dinh doc tinh ban trucmg diin cua vien nang Hoat huyet an nao tren thuc nghiem (2) Danh gia tac dung cdi thien tri nh& cua vien nang Hoat huyet an nao(HHAN) tren thuc nghiem. Phtrffng phdp nghiin cuu: Nghien cim doc tinh ban trirong dien theo hu&ng dan cua WHO, tho udng thudc thu Id Hoat huyit an nao liiu 0,36 g/kg/ngdy va lieu 1,08 g/izg/ngay trong 8 ludn lien tuc. Ket qua: Hoat huyit an nao khong lam dnh huang den thi trgng, chitc nang tgo mdu cua tho; khong lam thay doi chicc nang gan, chuc nang than; khong gay ton thiccmg ve hinh thai khi quan sat dai the vd vi the cdc co quan gan, than tho. Thuoc thu Hoat huyet an nao cd 2 lieu 0,72g/kh/ngay vd 2,16g/kglngdy tren chuot nhdt trang cung da the hen tac dung cdi thiin tri nha tren mo hinh Morris water maze thong qua khd nang lam gidm thai gian vd quang duang tim thdy chdn di, lam tdngphdn tram thai gian bai cua chuot trong 'A be c6 chdn di.
Kit luan: Vien nang HHAN an todn vd co tdc dung cdi thien tri nha tren thuc nghiem Tir khoa: Hoat huyet an nao, d^c tinh, cai thien tri nhor, Morris water maze, thirc nghiem.
Summary
SUB-CHRONIC TOXICITY AND MEMORY-ENHANCING ACTIVITY OF HOAT HUYET AN NAO IN EXPERIMENTAL ANIMALS
Objectives: 1) To study on the sub-chronic toxicity of "Hoat huyet an nao' capsules (HHAN) in experimental animals. 2) To evaluate the memory-enhancing effect of HHAN In experiment Metltods: In sub-chronic toxicity study, groups of 10 Newzealand white rabbits were administered orally HHAN at doses of 0,36g/kg/day and 1,08 g/kg/day for a period of 60 days. Results: HHAN did not cause damage to general state, hematopoietic, liver and kidney function in rabbits. No histological changes were observed in organs of rabbits. Effect of HHAN on memory improvement in mice was evaluated using Morris water maze model HHAN at doses of 0,72g/kg/day and 2,16g/kg/day significantly improved memory ofm ice, indicated by decrease in escape latency during training and increase in time spent in target quadrant during retrieval. Conclusions HHAN capsules is safe and had memory-enhancing activity in experiment.
Keywords: Hoat huyet an nao, toxicity, memory enhance, Morris water maze, experiment.
Dia Chi lien h^: Tr^n Minh HiSu - Benh vien Y hoc c5 truyen TW Ngay nhan bai: 20/10/2015 Bja chi email: [email protected] Ngay phan bien: 09/11/2015 Phan bien khoa hpc: PGS.TS. Vu Nam Ngay dang bao: 18/12/2015
• TTiS.BS; ** TS.DS; *** PGS.TS, **** BS;
Journal of research in viet nam traditional medicine and pharmacy I. DAT VAN DE
Tai biSn mach.mau nao (TBMN) la mot van de lan cua Y hoc cac nirac trong nhieu thap ky qua. Benh co the gay tu vong nhanh chong ho^c de lai nhieu di chung nang ne nhu liet van dong, roi loan ca tr6n....NhGng nam gan day, TBMN a nuac ta co chieu huong gia tang va tre boa, anh huong nhieu den chat lugng song va lam tang chi phi chiia benh gay ton kem cho gia dinh, xa hoi [1],[2],[3]. Bai thudc nghiem phuong "Hoat huyet an nao"
(HHAN) g6m 11 vi thu6c co tac dung hoat huyet boa u, khu phong trir dam, phu hop de diSu tri Nh6i man nao.
Tren co so tac dung dugfc ly hien dai cua cac vi thuoc va bien cbumg luan tri chiing tning phong hoac ban than hit toai, chung toi tien hanh nghien cmi tac dung dieu tri phuc hoi chiic nang benh nhan NMN sau giai doan d p ciia bai thu6c HHAN. Tnioc khi danh gia tac dung cua bai thuoc tren lam sang, chung toi tien hanh danh gia bai thu6c ve tinh an toan cung nhu tac dung tren thirc nghiem.
Mvtc tieu:
1. Xdc dinh doc tinh ban trw&ng dien cua vien nang Hoat huyit an nao tren thuc nghiem
2. Danh gia tdc dung cdi thien tri' nh& cua vien nang Hoqt huyit an nao tren thiec nghiem
U. CHAT LIEU, DO! TU*gfNG VA PHU'ONG PHAP NGHIEN CUtJ
2.L Chat lieu nghien cvu:
2.1.1 Bai thuSc HHAN: H3ng hoa, xuyen khung, sinh dia, nguu tit, dao nhan, dia long, dan sam, cam thao, xich thuac, duong quy, hoang ky; san xult tai TT Duoc BY YHCTTW, dong nang 0,5g/vien, lo 60 vien.
2.1.2. Hoa chat, dung cu phtfc vyi nghien c^u: cac hoa chat, may moc chuyen khoa
2.2. Doi tiTffng nghien cihi::
- Tho chung Newzealand White, long trang, trong lugfng 1,8-2,5 kg do Trung tam cung cap dong vat thi nghiem Dan Phucmg - Ha Tay cung cdp
- Chuot nhat trang chung Swiss ca 2 giong, thuan chung, khoe manh, trong luong 25 gam do Vien Ve sinh Djch te Trung uang cung c4p.
2.3. Phu-ong phap nghien ciru:
2.3.1. Boc tinh ban trir&ng diin (theo doi trong 8 tudn): Theo huang din ciia WHO
Tho duac chia lam 3 16, m6i 16 10 con, moi con nhot rieng mot chu6ng.
L6 chung: uong nuac cat 3ml/kg/ngay
L6 tri 1: uong HHAN lieu 0,36g/kg/ngay(7/eM co tdc dung tuang duong tren nguai, tinh theo he s6 3)
- L6 tri 2: u6ng HHAN hku 1,08 g/kg/ngay (gkp 3 \kn 16 tri 1)
Nghien citu YDHCT Viet Nam So 47-2015 Cdc chi tieu theo doi tru&c, sau 4
tudn, 8 tudn uong thuoc:
Tinh trang chung, th8 trong cua tho.
CT mau (BC,HC,TC,Hct,Hb), AST, ALT, Alb, Creatinin, Bil TP, Cholesterol TP.
- Mo benh hoc: sau 8 tulin u6ng thuoc, kiem tra nglu nhien cau true vi thi gan, than.
2.3.2. Tdc dung cdi thien tri nh&
tren mo hinh Morris water maze [4]
Thu nghiem tien hanh trong 6 ngay: Gd 1: huan luyen trong 5 ngay dau, Gd 2: ngay thu 6 thu nghiem kha nang hoc va nha ciia chuot, gom 5 16, moi 16 10 con:
- L6 1 (chung trang): Tiem mang bung nuac muoi sinh ly 0,lml/10g
- L6 2 (mo hinh): Tiem mang bung scopolamin lieu Img/kg, 0,lml/10g
- L6 3 (chung duong): Uong thu6c chung duong Donepezil lieu 2,4 mg/kg, 0,2 ml/lOg, 30 phiit sau tiem mang bung Scopolamin lieu 1 mg/kg, 0,lml/10g
- L6 4 (HHAN lidu thSp): U6ng HHAN lieu 0,72g/kg/ngay (lieu tuong duang 1dm sang), 30 phut sau tiem mang bung scopolamin lieu 1 mg/kg, 0,1 ml/1 Og
- L6 5 (HHAN li6u cao): Uong HHAN lieu 2,16g/kg/ngay (gap 3 lan 16 tri 1), 30 phut sau tiem mang bung scopolamin lieu 1 mg/kg, 0,lml/IOg
Chuot dirge tiem va cho uong thu6c trong 6 ngay lien tiep.
Cac chi tieu danh gia:
- GD 1: thai gian chuot tim thSy chan de ben do; chieu dai quang duong tim thay chan de ben d6, phhn tram thofi gian chuot a goc Vt co ben d6.
- GD 2: % thai gian trong 1 phut chuot trai qua trong 14 be truoc do dat chan de.
3.4. Xfr ly so lieu: Cac s6 lieu nghien cuu dugc xu ly th6ng ke theo phuang phap T-test Student, bieu dien dang: X± SD, s\x khac biet co y nghta khi p < 0,05.
III. KET QUA
3.L Doc tinh ban tru'o'ng dien 3.1.1. Tinh trgng chung: trong 8 tuan, tho a ca 3 16 hoat dong binh thuang, nhanh nhen, mat sang, long mugl, an uong tot, phan kho. Trong lugng tho a ca 3 16 deu tang. Dp tang trong lugng tho giua 16 chung va 16 4,5 khong co sir khac biet (p > 0,05).
3.1.2. Danh gia chuc nang tgo mdu:
Journal of research in viet nam traditional medicine and pharmacy No 47 - 2015 Bang 1. Anh hu-ong cua HHAN den so lu-tfng hong cau trong mau tho
Thoi gian
Tnldrc uong thuoc Sau 4 tuan p (tnioc - sau)
Sau 8 tuan p (tnrac - sau)
So lirong hong cau ( T/I) Lo chu-ng
4,90 ± 0,46 4,84 ±0,53
>0,05 5,20 ± 0,55
>0,05
Lo tri 1 Jc±SD 5,08 ± 0,50 5,04 ±0,67
>0,05 5,54 ±0,56
>0,05
Ldtri2 'x±SD 4,94 ±0,52 4,77 ±0,51
>0,05 5,06 ± 0,40
>0,05
P (T-test)
>0,05
>0,05
>0,05 Nhan xet: sau 4, 8 tuin uong HHAN, s6 luong HC khong c6 su thay doi voi
p >0,05. " — 3
% a
"a
1
12 10 8 6 4 2 0
Tnioc uong thuoc
Sau4 tudn
1
Sau tui n 8
• Chung - O L d t r i l
B L d t r i 2
g
e
1
40 -, 30 - 20 10
0 -
III
S3u4 t u ^ Sau 8 tuan• Oiung QLdtril BLdtri2
Bieu do 1. Anh hirong cua HHAN den ham luong huyet sac to
Bieu do 2. Anh hir&ng cua HHAN den hematocrit trong mau tho Nhan xet: sau 4, 8 tuan uong HHAN, luong Hb va Hct trong mau tho kh6ng co su thay doi vai p >0,05
Bang 2. Anh hvong cua HHAN den so lircnig bach cau trong mau tho
Thol gian
Tnroc uong thuoc Sau 4 tuan p (truoc - sau)
Sau 8 tuan p (truoc - sau)
So lu-ong bach cau (G/1) Lo chirng
Z ± 5 D 6,96 ±1,26 6,61 ± 1,84
>0,05 7,05 ±1,47
>0,05
Lo tri 1 'x±SD 6,15 ±1,29 7,03 ±1,76
>0,05 6,62 ±1,43
>0,05
Lo tri 2 J±SD 7,42 ± 2,47 6,76 ± 2,09
>0,05 6,90 ± 2,24
>0,05
P (T- test)
>0,05
>0,05
>0,05 Nhan xet: sau 4, 8 tuiin u6ng HHAN, so lucmg BC khong co svt thay d6i voi p>0,05.
Nghien elm YDHCT Viet Nam So 47-2015 Bang 3. Anh huang cua HHAN den so lutfng tieu cau trong mau tho
Thoi gian
Truoc uong thuoc Sau 4 tuan p (tnrac - sau)
Sau 8 tuan p (tnroc - sau)
So lugng tieu c^u (G/1) Lo chung
X±SD 311,80 ±64,94 250,90 ±97,94
>0,05 332,20 ±48,32
>0,05
Lo tri 1 'X±SD 297,50 ±31,93 246,70 ± 85,43
>0,05 295,20 ±51,77
>0,05
Lo tri 2 'X±SD 304,80 ± 23,86 271,70 ±41,78
>0,05 293,90 ±63,42
>0,05
P (T- test)
>0,05
>0,05
>0,05 Nhan xet; Sau 4, 8 tuan uong HHAN, so luong TC khong co sir thay doi voi p>0,05.
3.1.3 Anh hirdng ten chu-c nang gan, than:
^
£
60 40 20 0
Truocuong Sau4tuan S a u S t u ^ ttiudc
• C3iung Q L o t r j l B L d t r i 2
T f u 6 c u ^ g S a u 4 t u ^ S a u S t u ^ thuoc
•Chiitng Q L d t r i l Q L d t n 2
Bieu d6 3 & 4. Anh hu-OTig cua HHAN den ho?t do transaminase trong mau tho Nhan xet: sau 4, 8 tuSn u6ng HHAN n6ng do ALT, AST trong mau tho khong co su thay doi vai p >0,05.
- Anh huong cua HHAN len n6ng do Billirubin TP, Albumin va Cholesterol toan phlin: kit qua thu duac sau 4, 8 tulin u6ng HHAN, cac chi s6 tren co su thay d6i khong CO y nghia vai p >0,05.
lalffl
Trucrc Sau 4 Sau 8 uong tuka tuSn thudc• Chiing DLotri 1 • L 6 U i 2
Sau 4, 8 tuan uong HHAN, nong do Creatinin khong c6 su thay doi voi p >0,05.
Bieu do 5. Anh hirong ciia HHAN den nong do creatinin trong man tho
Journal of research in viet nam traditional medicine and pharmacy 3.1.4. Hinh dnh gidiphdu vi thi gan vd than (HE x 400)
Nhan xet: Hinh anh vi the gan, than tho binh thuang sau 45 ngay uong HHAN 3.2. Tac dung cat thien tri nhdr tren mo hinh Morris water maze Bang 4. Anh htfOTig cua Hoat huyet an nao den thoi gian tim thay chan de
L o t h i nghiem
L o l P n - N 2 * i
L6 2 P(2-N21 P (2.11-1 L o 3 P (2.N21-2
P13-2, L o 4 P14-N21
P(4-21 P 14-31 L6 5 P (5 - N2)
P 15-21 P15-31 P(5-4)
Thoi gian tim thay chan de (giay) Ngay 1 (Nhin
thay chan de) 38,7 ±11,0 97,0 ± 24,7
<0,00I 54,1 ± 14,5
<0,001 62,0 ± 18,5
<0,01
>0,05 59,7 ± 22,4
<0,01
>0,05
>0,C5
Ngay 2 X±SD 48,0 ±15,0 99,3 ±18,8
<0,001 67,5 ±15,4
<0,001 66,8 ± 12,3
<0,001
>0,05 61,0±18,8
<0,001
>0,05
>0,05
Khong nhin Ngay 3 X±SD 38,7 ±15,5
>0,05 87,2 ± 26,6
>0,05
<0,001 53,5 ± 13,6
<0,05
<0,001 62,3 ± 8,8
>0,05
<0,05
>0,05 55,0 ±15,2
>0,05
<0,01
>0,05
>0,05
hay chan de Ngay 4 X±SD 34,5 ± 8,5
< 0,001 81,3 ±23,3
>0,05
<0,001 49,2 ± 12,1
<0,0I
<0,00I 51,5 ± 10,9
>0,05
<0,01
>0,05 50,5 ± 12,7
>0,05
<0,0I
>0,05
>0,05
Ngay 5 J±SD 23,2 ± 6,6
< 0,001 80,9 ±23,4
>0,05
<0,001 33,4 ±11,2
<0,01
<0,001 37,6 ±7,1 -
<0,0I J
<0,001 ,
>0,05 i 33,7 ±10,8 •
<0,001 i
<0,01
>0,05
>0,05
Nghien ctm YDHCT Viet Nam
* (Ghi chu: N2. Kdt qua ngay thu 2)
NhSn xet: Chuot uong thu6c HHAN cr cac lieu lam sang gap 3, gap 5 ISn liSu lam sang CO tac dung cai thien tri nho. So sanh voi 16 chiing va 16 2 d6u co y nghia voi p<0,01
Bang 5. Anh huong ciia HHAN den quang duong tim thay chan de
Lo thi nghiem
L o l
P(l-N2)
L6 2
P(2 - N2) P ( 2 - l )
L6 3
P(3 - N2) P(3-2)
Lo4
P(4 - N2) P(4-2) P{4.3)
LBS
P(5 - N2) P(5-2) P(5-3) P(5-4)
Quang duffug tim chan de (met) Ngay 1
(Nhin thay chan de)
'X±SD 12,5 ± 2,5 20,1 ±7,2
<0,001 15,0 ±2,7
<0,05 14,1 ±4.42
<0,05
>0,05 14,5 ±3,0
<0,05
>0,05
>0,05
Khong nhin thay chan de Ngay 2
J ± S D 12,7 ± 6,4
25,0 ±6,7
<0,001 18,0 ±6,2
<0,05 18,5 ±5,0
<0,05
>0,05 18,4 ±5,4
<0,05
>0,05
>0,05
Ngay 3 Y ± S D 11,1 ±3,2
>0,05 23,1 ±6,1
>0,05
<0,001 15,8 ±5,7
>0,05
<0,01 16,5 ±5,2
>0,05
<0,05
>0,05 17,1 ±5,5
>0,05
<0,05
>0,05
>0,05
Ngay 4 X±S£>
9,7 ±1,7
>.0,05 22,2 ± 4,7
>0,05
<0,001 13,1 ±4,7
<0,05
<0,01 14,4 ±5.8
>0,05
<0,01
>0,05 12,5 ±4,1
<0,05
<0,05
>0,05
>0,05
Ngay 5 'X±SD ' 8,2 ±2,4
<0,05 19,9 ±6,1
>0,05
<0,001 8,9 ± 3,5
<0,01
<0,01 12,2 ±4.6
<0,01
<0,01
>0,05 10,4 ± 2,6
<0,01
<0,001
>0,05
>0,05 Nhan xet: Quang duong tim thay
chan de ciia chuot: a 16 chung sinh hoc gi^m tung ngay, tuy nhien su khac biet c6 y nghia th6ng ke 6 ngay
5 so vai ngay 2 (p<0,05). L6 m6 hinh dung scopolamin, quang duang tim thay chan de khong co su khac biet so voi ngay 2 a tat ca cac ngay
Journal of research in viet nam traditional medicine andpharmaiy nghien cuu 3, 4 va 5 (p>0,05). So
voi 16 chung sinh hoc, scopolamin lam keo dai quang duong tim thay chan de 6 tat ca cac ngay tu 1-5, sir khac biet co y nghia thong ke (p<0,001). Chuot 6 16 3 dung donepezil va 2 16 dung HHAN (16 4 va 5) deu nit ngan quang duong tim
thay chan de so vai 16 m6 hinh o tat ca cac ngay nghien ctiu (p<0,05 va p<0,01).
Mijc do tac dung giita HHAN liSu thip va lieu cao la tuang duong nhan va tuong duong voi 16 dung donepezil lieu 2,4 mg/kg 6 cac thai diem nghien ciiu (p>0,05).
Bang 6. Anh hirong cua HHAN den ti le phan tram thoi gian chuot boi trong VA be trird'c do dat chan de
Lo thi nghiem Lo 1 (Chiing sinh hoc)
Lo 2 (M6 hinh) Lo 3 (Chiing duong)
Lo 4 (HHAN thip) Lo 5 (HHAN cao)
% Tg boi trong % be CO chan de 27,2 ± 8,5
9,2 ±3,8 23,1 ±5,8 22,2 ± 7,5 21.5 ±4.5
p so vol 161
<0,01
>0,05
>0,05
>0,05
p so vffi 16 2
<0,01
<0,01
<0,01
p so vol 16 3
>0,05
>0,05
p so voi 16 4
>0,05 Nhan xet: Thofi gian boi cua chuot
16 2 trong % be co chan de so voi 16 chung sinh hoc co sir khac biet voi (p<0,01). Lo 4, 16 5 chuot uong HHAN CO tac dung lam tang tri nhof cua chuot. Chuot boi trong 1/4 be co chan de lau hon. So sanh voi 16 mo hinh CO sukhac biet vai p<0,01.
IV. BAN LUAN
4.1. Doc tinh ban tru'o'ng dien 4.1.1. Kit qua bdng 1,2,3 vd bieu do 1,2 cho thay: cac chi s6 TB mau ciia tho 6 ca hai 16 tri deu khong co su thay doi vai p >0,05. Kit qua nay
phu hop voi cac nghien cuu bai thuoc nghiem phuang truac day: Hoat huyet dan cua Tran Quoc Binh, Nhi chi thang cua Nguyen Boi Huong deu thay cac thu6c YHCT hau nhu kh6ng gay anh huang den chuc nang tao mau.
4.1.2. Anh hu&ng cua HHAN len chuc nang gan, than tho:
De danh gia muc dp t6n thuang te bao gan, nguoi ta thuong dinh luong nong dp cac enzym c6 nguon goc tai gan CO trong huyet thanh. Su tang nong do cac enzym nay thuong gan lien voi doc tinh cua thuoc do su buy
Nghiin c&u YDHCT Viit Nam So 47-2015 hoai te bao gan [5], [6]. Kit qua bi6u
do 3,4 cho thay sau 4, 8 tulin u6ng HHAN, a ca 16 tri 1 va 16 tri 2 hoat do ALT, AST, Albumin, Bil TP, Choles TP diu nam trong gioi han binh thuang. Dieu nay cho th4y thu6c HHAN an toan va khong gay doc voi gan tho tren thuc nghiem
Creatinin mau la chi tieu tin cay va quan trong hon ure mau, nen hien nay diing de danh gia va theo doi churc nang th|n. Ket qua bieu d6 5 cho thiy nong dp creatinin trong mau tho sau u6ng HHAN khong co su thay d6i so voi 16 chiing va giua hai thoi diem truac va sau khi uong thuoc thu (p>0,05)
4.1.3. Hinh dnh md benh hoc tho:
Giai phau dai the va vi the gan than la tieu chuan vang de danh gia t6n thuang 2 co quan chuyen hoa va thai trCr thuoc.
Tren tat ca cac tho nghien cuu, khong quan sat thay co thay doi benh ly nao ve mat dai the ciia cac co quan.
Hinh anh vi the gan, than kh6ng co sir khac biet giua 16 chung va 16 nghien cuu.
Cac k6t qua trSn da mot lln nua kh§ng dinh sir an toan ciia HHAN tren
thuc nghiem va la mot gpi y de co the diing thu6c tren lam sang trong cac nghien cuu sau nay.
4.2. Tac dung cai thien tri nho^
tren mo hinh Morris water maze Ket qua bang 4, 5 va 6 cho thay, tren m6 hinh Morris water maze, dung scopolamin {16 2) tren chupt nhat trang da lam keo dai thai gian va quang duang tim thay chan de, lam giam ty le boi trong VA be co chan de, qua do cho thay scopolamin da gay suy giam tri nha tren chuot nhat trang. Tren mo binh gay suy giam tri nho bang scopolamin, sau do dung donepezil lam thuoc chiing duong cho thay donepezil lieu 2,4mg/kg giam ro ret thai gian va quang duong tim thay chan de, lam tang ro ret phan tram thai gian boi ciia chuot trong '/4 be co chan de.Thu6c thu la HHAN ca 2 lieu 1,44 vien/kg/ngay va 4,32 vien/kg/ngay da lam giam thofi gian va quang duang tim thay chan de, lam tang phan tram thoi gian boi ciia chuot trong VA be co chan de, tac dung giua 2 lieu la tuong duong nhau va tuang duong voi donepezil lieu 2,4mg/kg.
Journal of research in viet nam traditional medicine and pharmacy
V. KET LUAN
1. HHAN kh6ng gay doc tinh ban tniong diin tren tho khi cho tho u6ng lieu 0,36 g/kg/ngay (liSu co tac d\ing tuong duong liSu ding tren nguoi) va liSu cao gip 3 ISn (1,08 g/kg/ngay) trong 8 tuSn lien tuc.
2. HHAN CO tac dung cai thien tri nha tren mo hinh Morris water maze, HHAN ca 2 li6u 0,72g/kg/ngay va 02,16g/kg/ngay da lam giam thoi gian va quang duong tim thay chan dS, lam tang phan tram thai gian boi ciia chupt trong VA be co chan de.
TAI LIEU THAM KHAO
1. Nguyen Van Bang (2000), Tai bien mack man nao, Nha xuSl ban Y hoc
2. Organisation for Economic Co-operation and Development (2002), Guideline for testing of chemicals: Acute Demal Irritation/Corrotion, OECD 404.
3. World Health Organization (2000), Worthing group on the safety and efficacy of herbal medicine.
Report of regional office for the
western pacific of the World Health Organization.
4. Varvel SA, Lichtman AH (2002), Evaluation ofCBl receptor knockout mice in the Morris water maze, J Pharmacol Exp Ther, 301(3), 915-924
5. Nguyen The Khanh, Ph?m Tir Duong (2001), Xet nghiem sCi dung trong lam sang, Nha xuat ban Y hoc.
6. Vii Dinh Vinh (2001), Huang dan sic dung cdc xet nghiem sinh hoa, Nha xult ban Y hoc, 115-287.
