O Nghien CLFU - Ky thuat
Nghien ciiu tac dung an than thirc nghiem ciia che pham sleepy care: Thong bao so 1
^<illycn Thj Minh Ngoc, Nguyen Thu HSng, t)inh Dai Do, Phting Thanh Hiro'ng, Nguyen Hoang Anh
Tnr&ng cigi hoc Dm/c Ha !\'6i
D a t v a n de
Mat ngu la roi loan t h i n kinh kha pho biln, gay hau qua v l syc khoe, tam iy, kinh te va c h i t lyong cu6c sing cho benh nhan va cho xa hoi Han e h l Idn nhIt cua viec sy dung cac thuoc an t h i n c6 nguon goc h6a duoc, nh6m du'oc iy quan trong trong dilu tn m i t ngu ia kha nSng gay le thuoc thulc va quen thulc Do d6, sy dung cac thulc va san phIm c6 nguIn g l c duoc iiSu hiSn dyoc coi la hydng tilp can bo sung va thay t h i cho cac thuoc CO nguon goo hoa d u o c ' ' ' Sleepy care, mot e h l phIm co nguIn g l c dyoc lieu g6m 4 thanh phan L-theanin, melatonin, cao valenan va cao common hops, da dyoc lu'u hanh tren thi tru'dng Viet Nam dudi dang thue pham ehu'c nang h i tro dieu tn mIt ngu Tuy nghien edu v l tac dung an thin cua tumg thanh phIn rieng biet trong o6ng thu'c da du'oc tiln hanh nhu'ng chya co nghien CU'U d l y dii nao ve tac dung ciia sy k i t hap 4 thanh phan trong c6ng thue eua che p h I m '^"^' Nghien edu nay du'oc thu'C hien vdi muc tieu danh gia tac dung an t h i n eua che pham sleepy care va tu'ng thanh phIn trong cong thu'C tren mot so m6 hinh thyc nghiem d chuot binh thydng n h i m cung cap them cac bing chdng v l tac dung du'OC ly cua c h i p h I m
D6i tu'O'ng va p h u ' c n g phap nghien CCPU
Nguyen lieu, hoa c h i t
C h i p h I m sleepy care, thanh,phIn cong thyc b a o g l m bot L-theanm 90-110 mg, bot melatonin 1.2-1.5 mg, cao valenan 45-55 mg, cao common hops 135-165 mg Diazepam (biet d y o c seduxen cua Gedeon Richter, Hungan), thiopental natn eua Rotexmedica, Ode, strychnin sulfat cua Cong ty co phIn Du'oc p h I m TW2
D6i tycvng nghien cii-u
Chuot n h I t t r l n g 2 g i l n g , chung Swiss albino khoe manh, 4 t u i n tuoi, k h l i l y o n g trung binh
20+2 g duoc cung d p bdi Vien Ve smh Dich t l trung yong Chuot d y o c chia 16 n g l u nhien 8- 16 con/16, nuoi trong d i l u kien nhiet d6 24±1°C, anh sang tu nhien, d u o c nu6i b i n g thdc an chuan, uIng nude ty do.
P h y c n g phap nghien CLPU
+ Diiu kien tiSn hanh: T i t ca cac thi nghiem d I u d y o e t i l n hanh trong phong neng biet, yen tTnh Thdi gian t i l n hanh thi nghiem trong khoang t y 8h-17h T r y d e khi t i l n hanh thi nghiem chuot d y o c lam quen vdi d i l u kien ph6ng thi nghiem trudc 1 gid. Chu6t d y o c u6ng thuoc trong 5 ngay lien tue Tien hanh cac thi nghiem vao ngay thy 5, sau khi cho chuot uong thulc 30 phut
+ Su" dung cac lidu sleepy care S C I , SC2, SC8, tyong yng vdi hlu b i n g 1x, 2x va 8x hlu ngoai suy ty Iilu sy dung tren ngudi Lieu cua tyng thanh p h I n L-theanm, melatonin, cao valerian va cao common hops d y o c ngoai suy t y lieu cua che pham toan p h I n va ly le p h I n tram cac thanh phan nay trong cong thyc. Lieu cua diazepam la 2 mg/kg the trong chuot Cac t h u l c thy d y o e pha trong CMC 0.5% d l tao h i n dich cho chuOt u6ng
+ Danh gia tac dung an thin cua sleepy care tren chuot binh t h y d n g
- Mo hinh leo day (m6 hinh Gnp) theo mo ta cua Jamal va c6ng sy '^' Chuot d y o c dat nhe nhang len day b i n g 2 chan trydC- Hoat dong cua chuot dyac quan sat trong 5 giay va d y a c danh gia theo thang d i l m : 0 diem ehu6t rai, 1 d i l m : chu6t giy nguyen ty the, 2 diem: chuot giy nguyen t y t h i va e l g i n g treo len, 3 d i l m chuot c6 g i n g treo len bang 2 chan trydc vdi 1 hoac ca 2 chan sau, 4 dilm, chu6t treo len bang ca 4 chan va q u i n duoi. 5 d i l m . chuot chay duoc tren day
- M6 hinh chy thap nang cao (elevated plus maze) theo m6 ta Emamghoreishi va cong sy''''
TAP CHi DUOC HOC - 01/2012 (SO 429 NAM 52)
© N g h i e n CLFU - Ky t h u a t
Dung cu thi nghiem dyoc cau tao la hinh chu' thap gom 2 tay kin kich thudc 30 x 5 x 15cm (dai x rong x cao), 2 tay hd kich thudc 30 x 5 x 0,2 cm, 2 tay kin va hd dat vu6ng g6c vdi nhau tao ra khoang trung tam co kich thu'dc 5 x 5 cm B6 dung cu duoc dat each nen nha 45cm. cyong d6 chieu sang trong thdi gian lam thyc nghiem dooc duy tri bang m6t bOng den 60V\/ Chuot duac dat nhe nhang vao viing trung tam, mat hydng v l canh tay hd, sau do du'oc ty do di chuyin kham pha trong 5 phut Chuot duoc coi la d trong 1 canh tay khi ca 4 chan d I u n i m trong canh tay do Sau khi tien hanh thi nghiem vdi moi chu6t, dung cu thi nghiem duoc lau sach trydc khi tien hanh voi chuot tilp theo de tranh lyu giO' mui Chi tieu danh gia ghi nhan s l lln ra vao va thdi gian lyu lai d cac vung canh tay (dong va/hoac md) eua chuot
- Anh hydng tren thdi gian ngu do thiopental xac dinh-theo m6 ta cua Db Trung Oam va D6 Thi Phu'ong'^' Chuot d y a c tiem tinh mach duoi thiopental hlu 40mg/kg Dat chuOt n I m nghieng, thai gian ngu do thiopental duo'c tinh tu' khi chuot b i t d I u ngu d i n khi chuot co phan xa lat minh trd day
Tac dung d l i khang co giat gay ra bdi strychnin theo mo ta ciia D l Trung Oam va 0 6 Thi Phyong'^' Chuot d y o c tiem d y d i da strychnin sulfat lieu 1 mg/kg Quan sat dap yng eua chuot Tren chu6t eo giat, ghi nhan ty le phIn tram chu6t co giat va thdi gian t i l m tang k l tu' khi tiem strychnin d i n khi chuot co giat Tren chu6t chet. ghi nhan ty le phan tram chu6t ehit va thdi gian t i l m tang ke t y khi tiem strychnin den khi ehuot co giat va c h i t
X y ly so lieu
DLT lieu duoc luu try, phan tich b i n g phan m i m SPSS 16 0 va du'oc bieu d i l n d u d i dang gia tn trung binh ± d6 lech chuIn (X ± SD) hoac ty le.% Qua phan tich k i l m chuIn Kolmogorow- Smirnov;/ cho thIy mot s l d y lieu khdng tuan theo phan b l chuan Vi vay m l u khong tuan theo phan b6 chuIn va cac mau ma b i l n co tinh chat khong lien tue d y o c k i l m dmh b i n g test
Bang 2: Anh huung cua sleep care len thai gian nang cao
phi tham so Kruskal - Wallis sau do la Mamn - Whitney U test d l so sanh s y khac biSt giua cac 16 Vdi cac m l u tuan theo phan bo chuan, sy dung k i l m dmh One-v/ay Anova vdi hau kiem (post-hoe) Dunnette T3 d l so sanh sy khac biet giya eac 16 So sanh ty le d y a c thyc hien bing k i l m dmh x^ S d khac biet d y o c coi la co y nghia thing ke khi p < 0,05
Ket qua nghien cu'u
Tac dung ciia sleepy care tren mo hinh leo day
Kha nang bam gid tren day cua chu6t d cae 16 d y o c trinh bay d bang 1 K i t qua cho thIy, sleepy care vdi hlu S C I co xu hydng !am giam kha nang bam giy tren day cua chuot nhyng sy khac biet chya dat mdc co y nghTa thong ke (p>0,05) a hlu cao han (SC2, SC8), sleepy care lam giam kha nang bam giy tren day cua ehu6t so vdi 16 chyng (p<0,05), SC8 c6 tac dung tyong dyong diazepam (p>0,05)
Bang 1: Anh hwong sleepy care len kha nang bam giij' tren day cua chuot
1.6 Nacr Diazepam
S C I ' SC2'
SC8*
So chuot/lo (n) 10 ,= 10
10 10
10
Biem 3,10 + 0,57 1,10±1,10 2,40 ± 0,70 2,40 ± 0,52
1,90 ±0,57 Pi.:
P I : Pi-
Pi-:
P2!
P
. = 0,000 ,= 0,052 rt = 0,029 1 = 0,011 i= 0,001 i = 0,105 Tac d u n g cua sleepy care tren mo hinh c h y t h a p nang cao
Diazepam va sleepy care d cac muc hlu S C I . SC2, SC8 c6 tac dung lam giam thdi gian chuot lyu lai d tay kin va tang thdi gian lyu lai d tay hd so vdi 16 chyng (p < 0,05). Sleepy care d cac mu'c lieu SC1 lam giam thai gian luu lai tay kin, tang thdi gian lyu lai tay hd tuong dyong diazepam (p>0,05) Sleepy care d hlu SC2, SC8 lam tang thdi gian lyu !ai d tay hd tyong dyong diazepam (p>0,05), giam thdi gian lyu lai tay kin it hon diazepam (p <0,05)
Iwu lai d tay kin va tay ho tren mo hinh chCr thap
n Th&i gian tay kin (giay) 10 231,30 + 42,94
9 132,44 ±22,89 P p i : = 0.000
Thai gian tay ha (giay) 8,70 + 11,38 48.56 + 34,40
P pi.2=0,001 149,10 + 27,20
P;3=0 000 ,400
p,-i=0.015 P;-3=0,211
T.\P CHi n i OC H O C - 01/2012 (SO 42') NAM 52)
O Nghien CLFU - Ky thuat
Thai gian tay kin (giay) Th&i gian tay ha (giay)
1 5 5 , 9 0 ± 17,01 p . - : ^ 0 , 0 0 0 p2^ = 0 . 0 2 2
p , ^ = 0 , 0 1 5 P2-1^0,400
SCB'' 1 8 0 . 2 0 + 4 6 , 5 6 pi-5^
92-i-
^ 0 , 0 3 4
. 0 . 0 4 3 28,0 + 20,92 p i i = 0 0 1 5
p : ' ; = 0 , 1 8 2 Anh hu'dng tren t h d i gian ngii do thiopental
Anh hydng eua sleepy care d cac mdc hlu khac nhau tren thdi gian ngu do thiopental d u o c tnnh bay trong hinh 1 6' 16 chu'ng dung n y d c mu6i smh ly, thdi gian ngu trung binh cua ehu6t ia 9 04 phut Khi dung sleepy care vdi I i l u thap (SCI SC2), thdi gian ngu do thiopental tang len
1,5-2 Ian (p < 0,05) a hlu cao hon (SC8) co xu hydng keo dai thdi gian ngu do thiopental ciia ehu6t so vdi 16 chyng nhyng s y khac biet chya dat mdc cO y nghTa t h i n g ke (p > 0,05) Sleepy care d cac mUc Iilu S C I . SC2 c6 tac dung keo dai thdi gian ngu do thiopental tu'ong d y y n g nhau ( p > 0,05)
Hinh 1: Anh hwong cua sleepy care len thdi gian ngu do thiopental Tac dung doi khang tren co giat do strychnin
Tac dung doi khang tren co giat do strychnin cua sleepy care d cac mu'C h l u khac nhau d u o c trinh bay trong bang 3 O tat ca cac 16, 100% ehuot d I u co giat sau khi tiem strychnin Sleepy care Iilu SC2, SC8 e6 xu hu'dng lam tang ty le s i n g , giam ty le c h i t so vdi 16 ehdng nhu'ng s u khac biet chu'a dat mu'c eo y nghTa
thong ke (p>0.05) Tren thdi gian tiem tang co giat, sleepy care d cac muc lieu SC2, SC8 co tac dung keo dai thdi gian tiem tang co giat ciia chu6t so vdi 16 chung (p<0,05) Sleepy care lieu S C I cO xu hydng keo dai thdi gian t i l m tang eo giat eua ehuOt so vai 16 chyng nhyng sy khac biet chya dat mdc co y nghTa t h i n g ke (p>0,05)
Bang 3: Anh huong cua sleepy care len ty le chuot co giat. chet va thai gian tiem tang co giat do liiu cao strychnin
L6
NaCI
S C l '
S C 2 '
S C 8 "
n
12
10
11
11
Ti le chuot
(%)
1 0 0 100
1 0 0
100 Chuot CO zo giat
giat Th&i gian tiem tang (phut)
6 , 4 7 + 1,02
7,92 + 3,00
10.49 ± 2 , 9 7
9 , 1 2 ± 2 , 9 0 P th&i gian tiem tang
pi-2 = 0,418 P:-3 = 0,061
Pi.3 = 0,003 Pi-i = 0,013 P3-. = 0,478
Chuotchet Ty le chuot chet
(%)
2 5
3 0
9
9 Pi P3 P2- P'- P3 Pi
p
= 0 , 5 8 3
= 0 , 2 5 6
= 0 , 2 5 6
= 0 , 3 2 9
= 0,762
= 0,329 Tac d u n g an than cua ti>ng thanh phan
trong che pham sleepy care Mo hinh chu' thap nang cao
So vdi 16 chu'ng u6ng nu'O'C muoi smh iy.
melatonin lam tang thoi gian lyu lai d tay hd.
valerian lam giam thdi gian lyu lai d tay kin, tang s6 Ian ra vao cung nhy thdi gian lyu lai d tay hd con common hops lam giam thai gian lyu lai tay kin va tang thai gian luu lai d tay hd Tac dung lam tang thoi gian lu'u lai d tay hd eua melatonin
I V i ' C l l i n i ] O C H O C - ' " ' 2 0 1 2 ( S O 4 2 9 NA.M 52)
© N g h i e n CLFU - Ky t h u a t
va common hops tuong dyong sleepy care va diazepam Rieng valerian s l l l n vao tay hd tu'ong
dyong diazepam va thdi gian lyu lai d tay nay tyong duong vdi sleepy care va diazepam.
i i;' I l l H l
Hinh 2: Tac dung cua sleepy care va cac thanh phan trong cong thirc tren sd Ian vao tay ho (tren. trai). thoi gian luu lai tay ho (tren. phai) va thai gian Iwu lai tay kin (dwoi) Tren th&l gian ngu do thiopental' Trong
cae thanh phIn cO trong c6ng thdc chi eo valerian c6 tac dung keo dai thdi gian ngu do thiopental ro ret han so vdi nhom chung va du'ong vdi che p h I m toan phan Cac thanh phan con lai (L-theanm, melatonin va Common hops) CO xu hudng keo dai thdi gian ngu do thiopental so vdi 16 chung nhu'ng su' khac biet chya dat mCrc co y nghTa th6ng ke (p>0,05) (bang 4)
Bang 4: Anh hudng cua tu'ng thanh phin trong cong thirc len thai gian ngu do thiopental
Thai gian ngu do thiopental (phut)
scr
L-theanin""
8.20 i 5,41 j
2,24 1,83
= 0,000
= 0,247
= 0,010
= 0.428 Chops' 7 5,58 + 1,54 p,.r,= 0.169
Ban luan
De tao CO' sd cho viec ap dung che pham sleepy care trong ho tro dieu tn m I t ngu, trong khuon kho cua nghien cii'u nay, chung toi lu'a chon mot so mo hinh kinh d i l n d l danh gia tac dung an than thyc nghiem cua che pham bao gom mo hinh leo day (tac dung an than, gian
co), mo hinh chy thap nang cao (tac dung giai lo), d6i khang co giat gay ra bdi strychnin (tac dung u'C e h l t h i n kmh trung yong), anh hydng tren thdi gian ngu do thiopental (tac dung an thin) '^' Tren cac m6 hinh nay, e h l pham sleepy care da the hien tac dung an t h i n th6ng qua tac dung gian co lam giam kha nang bam giu' tren day, tang s6 l l n vao va lyu lai d tay hd dong thai giam thoi gian iu'u lai d tay kin, d l i khang co giat gay ra bdi strychnin, keo dai thdi gian ngu do thiopental cua dong vat thyc nghiem Tac dung an than cua sleepy care phu thuoc vao hlu s y dung Vdi mdc lieu tyong dyang vdi Iilu s d dung tren ngydi (SCI), sleepy care co xu hyong t h i hien tac dung an t h i n tren tat ca cac mo hinh tuy nhien tac dung t h i hien r5 ret nhat tren mo hinh chy thap nang cao va keo dai thai gian ngu do thiopental Tac dung gian co va d l i khang co giat gay ra bdi strychnin cua c h i p h I m chi the hien khi s d dung d mdc lieu cao hon SC2, SC8
O l tim hieu vai tro cua tdng thanh p h I n trong cong thue lien quan den tac dung an than cua c h i pham sleepy care, chung t6i da danh gia tac dung cua cac thanh phan nay tren hai m6 hinh chy thap nang cao va keo dai thai gian ngu cua thiopental vdi lieu tyong du'ong co so sanh vdi che p h I m loan p h I n K i t qua cho thIy melatonin, valenan, common hops vdi mdc hlu
T A P C H i D L O C HOC-01/2012 (SO 42') N ^M 52)
© N g h i e n CLFU - Ky t h u a t
tuong dyong vdi lieu s d dung trong che pham toan phan co tac dung an than, giai lo thdng qua viec lam tang s l Ian vao va thai gian lyu lai d tay hd cijng nhy giam thdi gian lyu lai d tay kin tren mo hinh chO' thap nang cao so vdi nhom chumg Tac dung nay cua cac thanh phan tuong du'ong vol tac dung ciia e h l pham toan p h I n L- theanin chu'a the hien tac dung tren m6 hinh nay Khi danh gia anh hu'dng len thdi gian ngu do thiopental, ehi eo valenan t h i hien tac dung keo dai ro ret thai gian nay so vdi nhom chdng trong khi do cae thanh p h I n con lai e6 xu hu dng keo dai thdi gian ngu nhyng chya dat mu'c co y nghTa t h i n g ke Vi vay, co t h i so bo ket luan valerian la thanh p h I n chinh tao nen tac dung an than ciia sleepy care Cao valerian ehiet x u l t tu' r l cay Valenan officinalis L da du'oc s d dung rOng rai trong dieu tn m I t ngu va giai lo au Acid valeric ia thanh phan hoa hoc chinh va cung la thanh phan quan trong tao nen tac dung an t h i n ciia valerian d y o c gia thuyet do iam tang n l n g d6 c h i t dan truyin ye e h l GABA d than kmh trung u'O'ng '^'
Ket luan
1 Sleepy care eo tac dung gian eo' (mo hinh leo day), giai lo (mo hinh chu' thap nang cao, an thin (keo dai thdi gian ngii do thiopental) va do c h i than kmh trung y a n g ( d l i khang vdi co giat gay ra bdi strychnin) Tac dung an t h i n ciia sleepy care phu thuoc vao Iilu sii' dung Sleepy care vdi mii'c lieu b i n g 2 ian I i l u d i l u tn tren ngu'di (SC2) t h i hien tac dung an t h i n tren t i t ea cac m6 hinh tren
2 Valenan la thanh phan co tac dung an than r6 ret nhIt trong s l 4 thanh p h I n eua sleepy care t h i hien d tac dung lam tang s6 l l n ra vao va thdi gian lu'u lai d tay h d dong thoi giam thdi gian lyu lai d tay kin, keo dai thdi gian ngu do thiopental Cac thanh phan con iai (L- theanin. melatonin, cao common hops) cung t h i hien tac dung d m6 hinh chu' thap nang cao tren m6t s l ehl tieu thdi gian lu'u iai d tay kin (Common hops), thdi gian lyu lai d tay h d (melatonin, common hops) nhu'ng chya iam thay d l i co y nghTa thdi gian ngu do thiopental
Summary
Sedative properties of Sleepy Care capsules (a combination of L-theanin melatonin valerian
extract. common hops extract) was experimentally investigated by various models Sleepy Care displayed in a dose-dependent manner muscle relaxant activity (Grip strength test), anxiolytic activity (elevated plus maze model), hypnotic activity (potentiation of thiopental sleeping time) and CNS depression activity (at high doses, antagonism of strychnine-induced convulsions) Valrian extract was the most potent component, which increased open arm entries and the time spent on the open arm in EPM. prolonged thiopental- induced sleeping time All together these results provided expenmental evidences for the use of Sleepy Care as complementary in treatment of sleep disorders
Tai lieu tham khao 1 Wheatley D, Medicinal plants for insomnia a review of their pharmacology, efficacy and tolerabilily, Journal of Phsychopharmacology. (2005) 19, pp 414 -421
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7 Emamghoreishi M, Khasaki M , Aazam M 'Conandium sativum evaluation of its anxiolytic effect in the elavated plus-maze", Journal of Ethnopharmacology, (2005) 96, pp 365-370
8 Do Trung Oam, Do Thi Phuong, "Tac dung an thin cua senin, bot alcaloid la sen". Tap chi Duoc hoc. (2006)368. tr 19-22
9 Vogel H G (Editor). "Drug discovery and evaluation Phannacological assay' 2'"' edition, Springer-Verlag Berlin (2008)
