Nghien cicu YDHCT Viet Nam So 41 2014
x A C D I N H D O C T I N H C A P v A T A C D U N G H A G L U C O S E M A U
• • • •
C U A C A O L O N G SD T R E N T H U C N G H I E M
Studies on a c u t e toxicity a n d the effects of e x t r a c t SD on t h e p l a s m a glucose level on e x p e r i m e n t a l m o d e l
* Nguyin T r i n Thj Giang Huong ', ** Pham Thi Van Anh ', *** T r i n Quynh Trang \
**** Hoang Thanh Loan ^ ***** Nguyen Kilu Van ', ***** Dinh Quang Trucmg ' ' Tneang Dgi hoc Y Ha Noi, ' Trudng Dai hoc Duac Ha Noi
Tom Tat:
Xdc dinh doc tinh cdp vd tdc dung ha glucose mdu ciia cao long SD tren chuot nhdt trdng thuc nghiem. Cao long SD khong co doc tinh cdp, chua xdc dinh duac LDm cua thuSc thu tren chuot nhdt trdng bdng dudng udng theo phuang phdp Litchfield - Wilcoxon. Thuoc thu a lieu 10 g/kg vd 20 g/kg cd tdc dung ldm hg glucose huyit tren chuot binh thudng vd chuot bi gdy ddi thdo dudng bang alloxan.
Tir khoa: cao long SD, doc tinh c§p, ha glucose mau, thuc nghiem.
Summary
S T U D I E S O N A C U T E T O X I C I T Y AND T H E E F F E C T S O F E X T R A C T SD ON T H E P L A S M A G L U C O S E L E V E L O N E X P E R I M E N T A L M O D E L
The objective of the study was to evaluate the acute toxicity and the effect of extract SD on the plasma glucose level on mice. At lOg/kg bodyweight, extract SD had no acute toxicity and the
median lethal dose for oral route was not known. Extract SD with a dose of lOg/kg bodyweight and 20g/kg bodyweight, correspond to half of the clinical dose and the clinical dose had significantly reduced plasma glucose level on normal mice after oral route daily in a 4 week consecutive course.
The extract SD had significantly reduced plasma glucose level on alloxan induced diabetic mice after oral route daily during 20 days.
Keyword: extract SD, acute toxicity, reduction in plasma glucose levels, experiment.
Bja chi lien he: Pham Thj Van Anh - Truang Dai hoc Y Ha Noi Dja chi email: [email protected]
Phan bien khoa hoc: PGS.TS. Tran Quoc Binh
* PGS IS' **TS' *** SVY6" **** ThS' ***** KTV
Ngay nhan bai: 02/5/2014 Ngay phan bien: 16/5/2014 Ngay dang bao: 27/6/2014
Journal of research in viet nam traditional medicine and pharmacy No 41 2014
I. DAT V A N D E
Dai thao duong la mgt benh r6i loan chuyen hoa, la nguyen nhan gay tu vong hang dau tren thi gior [1].
Ben canh cac nhom thu6c dieu tri dai thao duorng dang dugc sii dung rgng rai tren the gioi nhu insulin, sulfonylure, biguanid ..., nhieu tac gia da di sau tim hieu tac dung ha glucose mau ciia cac cay con thu6c c6 san trong dan gian vi uu diem la ngu6n dugc lieu san co, de sii dung, gia thanh re, it tac dung kh6ng mong mu6n.
Thu6c thii SD g6m 6 vi: tri mau, sinh dia, cat can, mach m6n, hoang ky song, thuong truat, trong do sinh dia la vi chu cua bai thuoc. Theo y hgc c6 truyen bai thu6c co c6ng nang: Duong am, sinh tan, chi khat, ich khi. Chu tri: Nguoi am hu, hao khat, gay yeu, met moi, tao bon. Tai Viet Nam, cac nghien ciiu ve ve tac dung ha glucose mau rieng re ciia cac loai dugc lieu tren chuot binh thuong va chuot dai thao duong typ 1 cho ket qua t6t, tuy nhien chua co nghien ciiu nao danh gia sir ph6i hgp ciia cac
F
dugc lieu tren trong bai thu6c theo nguyen ly ngii hanh ciia Y hgc c6
\ F
truyen. Nghien ciiu dugc tien hanh voi muc tieu:
- Xdc dinh dgc tinh cap cua cao long SD
- Ddnh gid tdc dung ha glucose mdu cua cao long SD tren chupt
F
nhdt thirc nghiem
II. C H A T L I E U , D O I T U O N G V A
PHUONG P H A P N G H I E N C U U
F
2.1. Chat lieu nghien cuu.
F
2.1.1. Thuoc nghien ciru
* Thu6c tbii dugc bao che dang cao long tai khoa Dugc hgc C6 truyen, Dai hgc Dugc Ha Ngi.
2.1.2. Hoa chdt vd dicing cu nghien ciru
- Gliclazid vien nen 80mg do C6ng ty Dugc pham Ha Tay san xuat.
- Kit dinh lugng glucose hang Terumo, Nhat ban.
- Insulin Ig 10ml, 40UI/ml, biet dugc Scilin R ciia c6ng ty Bioton SA, BaLan.
Alloxan Ig 20g ciia Hang Bio Basic INC, Canada.
F
2.2. Doi tugng nghien cuu:
Chuot nhat trang chiing Swiss, khoe manh, ca hai gi6ng hgng lugng trung bmh 20 ± 2g do Vien Ve sinh dich te trung uong cung cap.
2.3. Phuong phap nghien cuu:
2.3.1. Nghien cieu dpc tinh cap
* Xac dinh LD50 cua cao long
> F F
toan phan SD tren chuot nhat trang bang duong u6ng theo phuong phap Litchfield - Wilcoxon va huong dan
•> F F
ciia T6 chiic Y te the gioi [5].
Truoc khi tien hanh thi nghiem, cho chuot nhin an qua dem. Timg 16
F F n, y
chuot nhat trang, m6i 16 it nhat 10
18
Nghien ciJcu YDHCT Viet Nam So 41 - 2014
con, dugc u6ng cao long toan phan
\ > >
SD theo lieu tang dan. Tim lieu cao
F F •y
nhat kh6ng gay chet chuot (0 %), lieu
F F F
thap nhat gay chet chuot hoan toan (100 %) va cac lieu trung gian. Theo
F
doi tinh trang chung ciia chuot va s6 lugng chuot chet 6 m6i 16 trong 72 gio. Tir do xay dung d6 thj tuyen tinh
9 F
de xac dinh LD50 ciia thu6c thii. Sau do tiep tuc theo doi tinh trang chung ciia chuot den bet ngay thii 7 sau khi u6ng thu6c thii lan dau.
2.3.2. Nghien ciiu tdc dung ha
F
glucose huyet tren chupt binh thir&ng
f F F
Chuot nhat trang ca hai gi6ng khoe manh dugc chia lam 4 16:
F F
L6 1: L6 chiing (u6ng nuoc cat) Lo 2: U6ng Gliclazid lieu 30 mg/kg.
F -y
L6 3: U6ng SD lieu 10 g dugc lieu/kg (Tuong duong V2 lieu lam sang)
L6 4: Uing SD lilu 20 g dugc lieu/kg (Tuong duong lieu lam sang)
Chuot dugc u6ng nuoc cat hoac
F -y
thu6c thii lien tuc trong 4 tuan vao cac bu6i sang. Truoc khi lay mau lam xet nghiem chuot dugc nhin doi qua
F r
dem, lay mau du6i chuot, tien hanh dinh lugng duong mau tai cac thoi diem to (chua u6ng thu6c), ti (sau u6ng thu6c thii 1 tuin), X^ (sau uIng thu6c thu 2 tuin), ts (sau uong thuoc thu 4 tuan).
2.3.3. Nghien cieu tdc dung ha glucose huyet tren chupt gay ddi thdo die&ng bdng alloxan:
Chuot nhat trang ca bai gi6ng khoe manh dugc chia lam 6 16:
F F
L6 1: L6 chiing (u6ng nuoc cat) L6 2: Tiem alloxan lieu
/ F
250mg/kg + u6ng nuoc cat
L6 3: Tiem alloxan lieu 250mg/kg + Tiem insulin lieu 1 Ul/kg
L6 4: Tiem alloxan lieu
F \
250mg/kg + Uong Gliclazid lieu 30 mg/kg.
L6 5: Tiem alloxan lieu
F y.
250mg/kg + U6ng SD lieu 10 g dugc lieu/kg (Tuong duong V2 lieu lam sang)
L6 6: Tiem alloxan lieu 250mg/kg + Uing SD lilu 20 g dugc lieu/kg (Tuong duong lilu lam sang)
F
Tat ca chuot 6 cac 16 dugc dinh lugng glucose mau lan 1 sau khi nhin doi qua dem. Cac 16 chuot tir 16 2- 6 dugc tiem alloxan lilu 250mg/kg.
Sau 72 gio tiem alloxan, dinh lugng glucose mau lan 2, chgn cac chuot bi dai thao duong co miic glucose khi d6i tren lOmmol/l dugc tham gia nghien ciiu. Cac 16 chuot dugc uIng nuoc cat hoac thulc thii trong 20 ngay sau khi gay mo hinh, dinh lugng glucose mau liic doi tai cac thoi dilm sau u6ng thu6c thu 10 va 20 ngay.
Journal of research in viet nam traditional medicine and pharmacy No 41 - 2014
III. KET QUA NGHIEN ClTU 3.1. Doc tinh cap
Sau khi u6ng thu6c thii cao long
F
SD, tat ca chuot trong cac 16 kh6ng CO hi6n tugng gi dac biet: an uIng, van dgng binh thuong, chuot kb6ng bi kho tho, di ngoai phan kh6, kh6ng thay xuat hien chuot chet trong vong 72 gio sau khi u6ng thu6c thu cao long SD. Mgt tuin sau khi uong
' ' -y F
thu6c thii tat ca cac chuot deu s6ng va kh6ng thay gi bat thuong 6 tit ca cac 16 chuot. Lieu t6i da chuot da dugc u6ng; 0,2ml/ lOg chuot trong 3
^ F /»> -y
lan lien tiep, m6i lan each nhau it
nhat 2 gio. Tuong duong 60ml/kg the trgng chuot. Nhu vay chuot da u6ng cao long toan phan SD voi lieu 1 lOg/kg the trgng chuot nhung kh6ng thay xuat hien dau hieu dgc tinh cap, kh6ng thay bat thuong gi sau Ituan ke tu khi u6ng thu6c thii lan dau. Vi kh6ng CO chuot chet a tat ca cac 16 nen chua xac dinh dugc LD50 ciia cao long SD tren chuot nhat trang theo duong u6ng bang phuong phap Litchfield - Wilcoxon.
3.2. Nghien cuu tac dung ha
F
glucose huyet tren chuot binh thuong:
Bang 1: Anh huo'ng cua cao long SD len nong do glucose mau cua chuot nhat trang F F
Lo chuot
•
Lo 1: U6ng nuoc cat Lo 2: Uong Gliclazid 30mg/kg
P(2-l)
Lo 3: Uong SD lieu 1 Og /kg
P(3-l)
Lo 4: U6ng SD lieu 20g/kg
P(4-l)
Nong do glucose mau mmol/1 (X ± SD^
T 6,44 ±
0,83 6,37 ±
1,27 6,56 ±
1,13 6,19±
2,29
^1tuan
5.98 ± 1,17 4,13
±1,18
<0,01 6,01 ± 0.92
>0,05 5,92 ± 0,63
>0,05
A 2 tuan
6,09 ± 0,63 3,28
±1,44
<0,001 6,93 ±
0,91
>0,05 6,85
±1,33
>0,05
^ 3 tuan
7,00 ± 1,19 3,34
±1,07
<0,001 6,56 ±
0,88
>0,05 5,55 ± 1,31
<0,05
|n = 10
* 4 tuan
6,84
±1,08 3,82 ± 0,88
<0,001 5,03 ±
1,01
<0,05 5,89 ± 0,84
<0,05 Ket qua bang 1 cho thay: cao long SD lieu 10 g/kg u6ng lien tuc sau 4 tuan lam n6ng do glucose mau giam xu6ng ro ret so voi 16 chiing (p<0,05). Cao long
•\ \ F
SD lieu 20 g/kg lam nong do glucose mau giam xu6ng ro ret so voi 16 chiing tai
t F F \ >
thoi diem sau uong thu6c 3 va 4 tuan (p<0,05). Miic ha glucose mau ca hai lieu gan tuong nhau. Tac dung ha glucose mau ciia ca hai lieu cao long SD deu kem hon gliclazid lieu 30mg/kg.
Nghien cihi YDHCT Viet Nam Sd41 2014 3.3. Nghien cuu tac dung ha glucose huyet tren chuot gay dai thao duong bang alloxan:
•> > '
Bang 2: Anh huong ciia cao long SD len nong do glucose mau cua chuot nhat trang gay dai thao duong bang alloxan
Lo chuot
L6 1 L6 2 L6 3 L6 4 L6 5 L6 6
Nong do glucose mau mmol/1 (X ± SD) T
6,08 ±1,48
>0,05 5,79 ±1,93 5,31 ±1,58
>0,05 6,48 ± 1,93
>0,05 5,86 ± 1,57
>0,05 6,05 ± 0,67
>0,'05
T72h
5,90± 0,58
<0,001 20,49 ± 7,07
19,99 ±4,39
>0,05 21,94 ±5,07
>0,05 20,14 ±5,43
>0,05 19,98 ±5,23
>0,05
TlO ngay
5,99 ±0,91
<0,001 20,01 ± 5,52 21,77 ±6,84
>0,05 9,78 ± 0,90
<0,01 15,35 ±2,58
>0,05 14,29 ± 4,97
<0,05
*^ 20 ngay
6,05 ± 1,04
<0,001 22,29 ± 6,37 22,57 ± 7,23
>0,05 8,30 ± 2,42
<0,001 15,64 ±3,58
<0,05 14,51 ±4,54
<0,05
P
P(l-2)
P(3-2)
P(4-2)
P(5-2)
P(6-2)
Ket qua bang 2 cho thay: Alloxan lieu 250mg/kg tiem mang bung sau 72 gio CO tac dung lam glucose mau tang cao, miic tang dat tren 3 lin so voi chiing va so voi thoi diem truoc khi tiem. Cao long SD 6 ca 2 lilu u6ng lien tuc trong 20 ngay lam ning do glucose mau giam xu6ng. Tac dung nay kem hon so voi insulin lilu
Idon vi/kg. Gliclazid lilu 30mg/kg kh6ng CO tac dung lam ha glucose mau tren m6 hinh gay dai thao duong thue nghiem bang alloxan.
4.1. Tren doc tinh cap
F F
Tren chuot nhat trang theo duong
F F \
u6ng, dung den lieu llOg/kg chua
F F
thay dau hieu dgc 6 chuot, kh6ng co chuot nao chet trong vong 72 gio sau
F F t F
u6ng thu6c. Day la the tich t6i da chuot CO the dung nap dugc, vi vay, chua xac dinh dugc LD50 ciia cao long SD tren chuot nhit tring theo
F
dudng u6ng.
4.2. Anh huo'ng cua cao long SD len nong do glucose mau:
Cao long SD lilu lOg dugc lieu/kg the trgng va 20 g dugc lieu/kg the trgng u6ng lien tuc trong 4 tuin
Journal of research in viet nam traditional medicine and pharmacy No 41 2014
da lam n6ng do glucose mau da giam mgt cach ro ret tir thofi diem sau u6ng thu6c 3 tuan. Cac nghien ciin ciia chiing toi ciing phii hgp voi cac tac gia khi nghien ciiu rieng tirng loai dugc lieu rieng nhu tri mau va sinh dia. Theo Nguyen Thi Huang Giang:
nghien ciiu tac dung ha duong huylt ciia mangiferin chiet xuat tir tri mau {Anemarrhena asphodeloides bunge)
r F F
tren chuot nhat trang cho thay boat chat nay o lieu lOOmg va 200mg/kg CO tac dung lam ha glucose mau tren chuot binh thuong [2]. Theo Dao Van Phan, ca tri mau va sinh dia deu c6 tac dung lam ha glucose mau tren chuot binh thudng nhung khong ro ret so vdi 16 chiing [4]. Nhu vay khi
ph6i hgp cac dugc lieu tren theo quan diem ngii hanh ciia Y hgc c6 truyen, trong do sinh dia la vi chinh d lieu bang Vl lieu du kien dung tren ngudi da CO tac dung lam ha glucose mau tren chuot nhat binh thudng.
V. KET LUAN
•
Chua xac dinh dugc LD50 ciia cao toan phan SD tren chuot nhat trang tren dudng uong.
Cao long SD lieu 10 g/kg/ngay va 20 g/kg/ngay lam giam n6ng do glucose mau d chuot nhat trang binh thudng.
Cao long SD lieu 10 g/kg/ngay va 20 g/kg/ngay lam giam n6ng do glucose mau d chuot nhat bi gay dai thao dudng bang alloxan.
TAI LIEU THAM KHAO 1. Ta Van Binh (2002), Nguai
y F
benh Ddi thdo dudng cdn biet, Nha xuit ban Y hgc Ha Ngi, tr 9 - 11, 13, 2 2 , 2 7 - 4 6 .
2. Nguyen Thi Huong Giang (2004), "Nghien ciiu tdc dung hg
F F
duang huyet cua Mangiferin chiet xuat tie Tri mdu {Anemarrhena asphodeloides bunge) tren chuot nhdt trdng", Luan van thac sy y hgc.
3. Hai Thugng Lan Ong "Le Huu Trac" (1995) Hai Thuang Y long Tdm ITnh, Nha xuat ban Y hgc.
4. Dao Van Phan, Nguyen Khanh Hoa, Nguyen Duy Thuan (2003), "Nghien ciiu sang lgc tac dung ha dudng huyet cua Sinh dia, Mong trau, Giao co lam va Tri mau".
Tap chi Nghien ciru y hoc 27, tr 1 - 6.
5. World Health Organization (2000), Working group on the safety and efficacy of herbal medicine.
Report of regional office for the western pacific of the World Health Organization.
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