Nghien CIFU - Ky thuat
Danh gia doc tinh cua Cereneed-caps tren dong vat thu'c nghiem v
Dinh Thj Tuylt Lan ', Tran Thanh Tiing ^"
' Trtfdng Dgi hpc Y Dugc Thdi Binh
• Trudng Dgi hpc Y Hd Npi 'E-mail: [email protected]
S u m m a r y
Acute and subchronic toxixity of Cereneed-caps were evaluated in experimental animals, the former by the Litchfield - Wilcoxon's method in Swiss mice and the latter by the guidance of World Health Organization in Wistar rats. The mice treated with Cereneed-caps at daily oral dose of 49.28 g/kg showed no symptoms of acute toxicity, LD^ was not identified. In rate, treatment with Cereneed-caps at daily doses of 334.8 mg/kg and 1674 mg/kg for 4 consecutive weeks exhibited no significant changes in general status, body weight and number of erythrocytes, hemoglobin concentration, hematocrit, mean corpuscular volume, number of white blood cells, percentage of types of white blood celts, platelets count, actMty of AST, ALT, total bilirubin, albumin, total cholesterol and creatinin levels. In addition, Cereneed-caps did not cause any change in histology of the liver and kidney in experimental rats. In conclusion, Cereneed-caps proved no acute toxicity in mice and showed no change in haematological indice, renal and hepatic function tests in rats.
Keywords: Cereneed-caps, acute toxicity, subchronic toxicity, experimental animals.
D0t van dl
Sa sut Iri tug vd bdnh Alzheimer Id nhdng nguydn nhan chlnh d i n d i n tinh frgng phy tiiuOe d ngudi eao t u i i . Bieu hign cua bgnh khdng nhung gdy suy giam fri nhd md cdn dnh hudng den nhieu ITnh v y c nhdn thue khdc nhau, kdm tiieo vdi nhdng r i i logn v l hdnh vi, gdy dnh hudng nghidm trpng d i n khd ndng hogt ddng hdng ngdy vd chat lupng eude sdng cua bdnh nhdn •''. Bdn cgnh nhdng phuang phdp d i thign frl nhd cua y hpc hign dgi, phuang phdp d i l u frj diing cdc ehe p h i m d ngudn gdc t u dupc ligu Id hudng nghidn cdu cGa nhilu nhd khoa hpc. Vidn nang cung Cereneed-caps la c h l p h i m c^ thdnh p h i n g i m 10 vj d u p c ligu gdm: hdng hoa {Flos Carthamitinctorii). xich I h u ^ c {Radix Paeoniae), duang quy {Radix Angelicae sinensis), xuydn khung (Rhizoma Ligustici wallichii), sinh dja {Rhizoma Rehmanniae), ehf xdc {FnjctusAurantii). sdi hd {Radix Bubleuri). nguu t i t {Radix Achyranthis bidentatae), cam tiiao (Radix Glycynhizae), bgch qud (Ginkgo biloba extract) ^.
Trong do nhilu vj dupc lidu nhu: bgch qud, duang quy. nguu t i t . . . da dupe chdng minh c6 tde dyng d l thidn tri nhd trdn ti^n tiiye nghigm va tren bgnh nhdn sa siit frl tijg P-*'. Tuy nhidn, cho d i n nay ehua ed nghidn cdu ndo v l tinh an todn vd hidu qud frdn tri nhd khi phdi hgp 10 vj ttiudc ndy vdi nhau.
o l xdc djnh tinh an todn eua Cereneed-caps,
chung tdi tiln hanh nghien cdu vdi myc tidu: Ddnh gia dpc linh d p vd dOc tinh bdn frudng dien eua Cereneed-caps tren dpng vgt thyc nghidm.
D o i tipgrng v a p h i f o n g p h a p n g h i e n CLPU Nguyen ligu
Thudc nghien cdu: Cereneed-caps do Cdng ty Cd p h i n Dupe Vgt t u Y t l Hai Duang san x u l t dgt tidu c h u i n c a sd.
- Moi vidn nang cdng chda: 15 mg cao kho bgch qua vd 450 mg cao kho hon hpp l u a n g duang vdi 9 g dupe ligu bao gdm: h i n g hoa (0,76 g), xich ttiuvc (1 g), duang quy (1 g), xuyen khung (1 g), sinh d\a (2 g), chi xae (0,6 g), sdi hd (0,6 g), nguu t i l (1,3 g), cam tiiao (0,74 g).
- Cach bdo d i e : Nghien mjn 2 logi cao, rdy l l y bOl, frpn bpt cao vdi tinh bdl, avieel, calel earbonat tiianh hdn hpp bOt kep, them ethanol 96% vd nudc tinh khiet de tgo khoi I m , xdt hgt, s l y vd sda hgt, bao tion (frdn vdl ta dupe gom bOt Talc, magnesi stearat) sau dd dupe ddng nang dgl c h u i n .
Dpng vat nghien c i h j
- ChuOt nhdt fring chung Swiss, 20-22 g do Vidn Vg sinh Djch le Tnjng u a n g cung d p .
- ChuOt d n g trdng ehung Wistar, 180-220 g do Tnjng tdm Cung d p dOng vdt thi nghidm Dan Phup'ng - Ha Tdy cung d p .
TAPCHI DU'<?C HQC -10/2016 (S6 486 NAM 56)
• Nghien cifu - Ky thuat
ChuOt dup'c nudi 5-7 ngdy tiirdc vd trong sudt thai gian nghien cdu d dieu kidn ptidng thi nghigm vdi d i y dCi thdc an vd nude udng tgi Bp m6n Dupe 1^ - T m d n g Dgi hpc Y Hd NOi.
Phuvng phap nghien ci>u Dgc tinh cap
Theo phuang phdp Litchfield - Wilcoxon '^.
Chudt n h i t b^ng, chia n g i u nhien ttianh tdng Id, moi 16 10 con, u i n g ttiudc tiid ttieo lieu tang d i n den till tich va ndng dp tdi da ma chupt d t h i dung ngp d u p c ChuOt d- mdi 16 se dupc theo ddi tinh b^ng chung, s y Uiay ddi hanh vi, nhjp thd, rdi logn ttiln tdnh, ndn, bai tilt... frong 7 ngdy sau udng thuie vd sd luvng ehuOt d e l Irong 72 gid.
Qua ty lg ehuOt c h i t Unh LD^^ d a ttiudc thu.
€>gc tinh ban truing diin
Thyc hlOn theo hudng d i n cua T i chdc Y t l T h i gidi •*•. ChuOt d n g I r l n g chia n g i u nhien Uidnh3ld, m d i l d l O c o n .
- Ld chdng: udng n y d c d t 1 ml/100 g/ngay.
- L6 tii 1: U i n g Cereneed-caps lieu 334,8 mg cao kh6 d u v c lidu/kg/ngdy (tuang duang vdi l i l u d y k i l n dCing eho ngudi, Unh theo hg s i 6).
- LO til 2; Udng Cereneed-caps vdi l i l u 1674 mg eao khd dupc lidu/kg/ngdy ( g i p 5 l l n Id bi 1).
ChuOt d u p t u i n g thudc 1 l l n vdo b u l l sang lidn tyc trong 4 h j I n .
Cac chi tidu tiieo ddi frudc, ti^ng, vd sau qua trlnh nghidn cdu d t i t d cac 16 nhu sau:
- Tinh trgng ehung, t h i trpng.
- Ddnh gid chuc phdn tgo mdu thdng qua sd lupng hdng d u , ttil tich frung binh hdng d u , ham lupng hemoglobin, hematocrit, sd lupng bgch d u , cdng Uidc bgch d u vd s i iupng tieu d u .
- Danh gid chuc ndng gan tiidng qua djnh lupng mOt sd chat chuyin hod trong mau: bllimbln lodn phan, albumin vd cholesterol loan p h i n .
- Danh gid mdc dO hiJy hogi l l bao gan tiidng qua djnh luvng hogt dp enzym frong mdu: ALT, AST.
- Ddnh gid chdc ndng tiidn tiidng qua djnh lupng nong dp creaUnIn huylt tiianh.
- c l u fruc dgi till vd vi till eua gan vd tiign.
T i t d cdc xdi nghidm fren d u p c danh gid d cae tiidi (Sim: frudc udng, sau 2 h j I n vd 4 t u i n udng ttiudc. Rieng hinh ttidi dgi ttil vd d u fruc vi fril gan, tiidn d u v c ddnh gia sau 4 t u i n u i n g thude d 30% sd chuOt moi Id.
Phdn t i c h vd xi> ly s d l[$u
Cdc sd lidu nghidn c u u d u v c x u ly thing ke ttieo phuang phdp t-test Shjdent vd test trudc - sau (Avant-apr^). Sd ligu d u p c bleu difin d u d i dgng: X ± SD. S y khdc bidt d ^ nghTa khi p < 0,05.
Ket qua nghien c i n i D p c t i n h cap
ChuOl d u p c u i n g Cereneed-caps vdi l i l u tdng dan d i n ndng dO vd t h i tich tdi da cho phdp vdi l i l u Id 49,28 g/kg ttil tn?ng chuOt ( g i p frdn 74 lln lieu d y k i l n dung cho ngudi). Theo ddi kh6ng thiy x u l t hidn tridu chdng b l l t h u d n g ndo trong suit 7 ngay sau udng thudc. Trong 72 gid d i u , khdng CO chuOt nao c h i t d tat d d e 16 nghidn cuu. Do do khong tinh d u v c LDj^ d a Cereneed-caps theo d u d n g udng.
Vi vdy theo Td chde Y t l till gidi, Cereneed-caps la ttiudc nguon gdc dupc ligu d tinh an t d n ^.
Dpc t i n h bdn t r u ' d n g d i l n
Tmh tr^ng chung vd sy thay d6i the frpng cda chugt
Trong thdi gian thi nghl0m, chuOt d d 3 Id hogt dpng binh thudng, nhanh nhgn, m i t sdng, ldng m u p l , dn udng tdt, phdn khd. Trpng lupng ehudt d i u tdng va khdng c6 s y khdc bigt gida 16 chdng va Id udng Cereneed-caps.
Dinh gia chirc ndng tpio miu
InrOc uAng Sau 2 tuJn uAnethu6c
• LAchi>ng D u t t r j l B u H Z B i l u do 1 ; Anh hu&ng cua Cereneed-caps
din s6 Itrpng hing ciu
B i l u d l 2: Anh hu&ng cua Cereneed-caps din s6 lupng b^ch du
TAPCHi Dl/<?C HQC- 10/2016 (S6 486 NAM 56)
w rvgriieh
Thoi gian Tnnfe uing thudc Sau 2 tuk p(trw!fc-sau) Sau4tu^
p(tnj^-sau)
CiPU - Ky thuat
Bang 1: Anh huong cua iochtjnq 526.10 ±138.63 655.001160,35
>0.06 564.30182,36
>0,05
<^eienee<U:aps den so /upng hiu cau bong miu chudt So hrong Siu cau (tall)
Lom<
606.00 ±146^5 652.00+147,18
>0.05 669.601164,02
>0.05
L6&T2 496,901122,65 582,60+126,18
>0,05 592,70199,00
>0,05
P (t-testShidentl
>0,05
>0,05
>0,05
K i t qua t d bieu dd 1,2 va bang 1 eho t h i y sau 2 t u i n vd 4 t u i n udng Cereneed-caps, sd lupng hdng d u , bgch d u va tieu d u d d 216 frj kh6ng d s y khde bigt d nghTa so vdi 16 chdng vd so sdnh gida cdc tiidi d i l m trudc va sau khi udng tiiudc ttiu ( p > 0,05).
Cdc tidi s i hemoglobin, hematocrit, cdng ttiue bgch d u d i u eho k i t qud tuang l y , khdng co s y khdc blOt d y nghTa thdng ke giua d c 16 u i n g Cemeed-caps vdi 16 ehdng vd so sdnh gida cac thdi d i l m trudc vd sau khi uong thudc tiiu (p > 0,05).
Danh gia m d c dg huy hopi te bio gan
Sau 4 tuin u ^ g l h u ^
Bleu do 3: Anh hu&ng cua Cereneed-caps den hopt d^ AST
Bieu do 4; Anh huvng cue Cereneed-caps din boat dp ALT
K i t qua d cdc bieu dd 3 vd 4 cho t h i y : sau 4 tuan udng Cereneed-caps, hogt dO AST, ALT trong mau chupt d d 2 16 trj d i u khdng e6 s y khac bigt d y nghTa so vdi 15 ehdng vd so sdnh giua hai thdi d i l m t r u d c vd sau khi udng t h u i e (p > 0,05).
Danh gia chirc nang gan
Bang 2: Anh hu&ng cua Cereneed-caps den nong dp bilinibin toan phan Thdi gian
TRK>C uing thuie Sau 2 t u ^ u ^ thu6c p(trudc-sau) Sau 4 tuin u6(ig thudc p{tiwic-sau)
LicltOng 13.4510,59 13,3610,42
>0,05 13,1610,3)
>0,05
Biliniliin toan f)hin(mmol/i) Ldlnl 13,5410,35 13,4110,36
>0,05 13310,40
>0,05
L817l2 13,2110^9 13,4910,49
>0.05 13.2410.40
>0,05
(t-teslSluilenI]
>0,05
>0,O5
>0.05
Ddnh gia chtic ndng thpn K i t qud d edc bdng 2 cho thdy sau 2 luan vd
4 l u i n u i n g Cereneed-caps cho I h l y ndng dp blllnjbln todn p h i n frong mdu chuOt d d 2 16 trj d i u khdng c6 s y khde bigt d y nghTa so vdi Id ehdng vd so sdnh giua hai thdi d i l m trudc vd sau khi udng tiiudc ttiu (p > 0,05).
Cht sd ddnh gid chuc ndng gan khde nhu cholesterol todn p h i n vd albumin frong mau chupt
cOng cho k i t qud tuong ty. Bieu dd 5: Anh huong cua Cereneed-caps din nong dp creatinin
T A P C H i D I Q C H Q C - 10/2016(S6486 N A M 56)
• Nghien ciju - Ky thuat
K i t qua d bieu dd 5 cho ttiay sau 2 U i l n vd 4 t u i n udng Cerwieed-caps, d d Id tn 1 va 16 trj 2, notxQ dO creatinin frong mdu chuOt kh6ng d s y ttiay ddi Wide bigt c6 y nghTa thdng kd so vdi 16 ehdng vd ttidi diem frudc nghidn cuu (p > 0.05).
Giai phiu vi thi gan (HE x 400)
Hinh anh giai phiu b^h Giai phiu dpi thi
Sau 4 t u i n udng ttiudc frdn t i t d d c d u p t ktwng quan sat t h i y d thay d i i bdnh ly ndo v l mgt dgi ttie cua d c ca quan tim, p h i i , gan, Idch, tuy, th0n va hg thdng tieu hod.
L6 chung Giai phiu vi thi th$n (HE x 400)
L o f r i l L6frf2
Loch img L d b i l Ldtrj2
Hlnh dnh vi t h i gan, ttign khdng eo bieu hidn ldn thuang, khing cd s y khde bigt giua 16 ehung vd 16 nghidn cdu.
Nh$n xdt Chung
Dd nghidn edu ddc tinh bdn frudng dien d a tiiudc Cereneed-caps ti^n chudt d n g frdng udng vdi l i l u 334,8 mg/kg/ngdy (hpong ducmg vdi lieu d y kien dung cho ngudi tinh ttieo hg sd 6) vd l i l u 1674 mg/kg/ngdy ( g i p 5 l l n lieu fren) tinng 4 friln. K i t qua, khdng ttiay ed anh hudng d i n cac tiidng s i huyet hpc nhu sd luvng hdng d u , ttie tieh trung binh h i n g d u , ham luvng hemoglobin, hematocrit, sd tupmg bgch d u , edng ttiuc bgch d u vd sd luvng tilu d u ; kh6ng anh hudng d i n d c ttrang sd hda sinh nhu bilimbin loan p h i n , albumin, cholesterol t d n p h i n , creaUnin. AST, ALT cGng nhu d u fruc dgi till vd vi ttil cua gan vd than chudt.
K i t l u ^ n
- M i u tiiude tiiu Cereneed-caps khdng gdy dpc tinh d p tidn ehuOt nhdt frdng khi eho chuOl
udng l i l u dung ngp tdi da Id 49,82 g cao khd dupc lidu/kg, g i p frdn 74 l l n l i l u d y k i l n dCing cho ngudi. Kh6ng cd chudt ndo c h i t , vl v | y kh6ng tinh dupe LD50 cua Cereneed-caps frieo dudng uing frdn chuOt n h i t trdng.
- M i u t h u i e t h u Cereneed-caps k h i n g gdy dOc tinh ban trudng d i l n trdn chupt d n g frIng kht udng l i l u 334,8 mg eao kh6 d u p c lidu/kg/ngdy vd l i l u 1674 mg cao khd d u v c ligu /kg/ngdy trong 4 tuan Ildn fa,ie.
T a i l i ^ u t h a m Ithdo
1. Phgm Thing (2010), B$nh Alzheimer vdcdcffii sa sOl tri tup khdc. Nhd xult bdn Y hpc. Hd NOi.
2. Walesiuk A., Trofimiuk E. and Braszko J. J.
(2005), 'Gingko biloba extiact dimini^es stiessHnduced memory deficits in rats", Pharmatxil. Rep.. 57 (2), 176- 187.
3. Cheng L L.. Chen X. N., Wang Y. et al. (2011), 'Z-ligustilide isolated from Radix Angelicae sinensis ameliorates the memory impairment induced by scopolamine in mice*. FHtOerapia. 82 (7), 1126-1132.
4. Ma A and Guo H. (1998). -EITect of Rarfjf