• Ngiiien ciju - Ky tiiuat
Tinh an toan va tac dung chong viem da cap tinh tai ch6 tren thyc nghiem
cua Item boi da Viemda AD bao che tir duffc lieu Iden long doin .
T? Van Binh*, LS Thanh Xuan^, Pham Thi Vdn Anh^, LS Thi Hai Y l n \ Nguyen Thi Hoa Hien', D6 Kim Chung^
' Tncang Cao dang Y te Hd Noi, ^ Truong Dgi hoc YHdNQi,
^ Binh vien YhQC Co truyen BQ Cong an D a t v a n d e
Bdnh vi@m da ca dia (VDCD) Id mpt benh ngodi da ph6 b l l n d Viet Nam va tren the gldi.
Cho den nay, viec d i l u tri VDCD vSn cdn ggp rat nhieu kho khan do can nguydn cua benh rdt phuc tgp, chu y l u Id d i l u tri trieu chung bang corticoid boi tgi cho ket hp'p vdi giu dm cho da, uong thuoc khdng hlstamin Hi vd d c vitamin nhung k i t qua khdng on djnh, tac dyng khong mong mu6n nhilu, ty id tdi phdt cao. Thuoc boi tgi cho chu yeu Id corticoid, neu bfli ddi ngdy gay teo da, gian mgch, doi vdi nhung BN bj tren didn rflng, boi se gay anh hudng d i n toan than. Cdc thuoc d i l u tri VDCD cd ngudn gflc tCp y hgc co truyen hien nay chua co tdc gia nao nghien cuu. Kien long ddm {Gentiana rigescens Franch.) Id 1 trong 4 loai long d d m , dup'c SCF dyng theo kinh nghiem de chdng ngua vd viem da cho benh nhdn. De gop phdn nghidn cuu thuoc y hpc co tmyen cd tdc dyng dieu trj VDCD, nghien cuu nay dug'c tien hanh n h i m ddnh gid tfnh an todn trdn da vd tdc dung chIng viem d p tinh tgi cho tren thyc nghiem cua che phdm d u g c bao che tit kien long ddm.
D o i t i r o n g v a p h u ' o n g p h a p n g h i e n CLPU
T h u o c n g h i e n CLFU
- Cao ddc c h l l t nudc vd cao dac chiet ethanol t u d u g c lieu kien long ddm (dgt tieu chudn Dugc dien Vidt Nam IV do Trung tam K i ^ nghiem Dugc vd My pham Ha Ngi xac djnh) do bg mon Dugc - Tm-dng CD Y te Hd NOi bdo che.
- Kem Viemda AD 5% va kem Viemda AD 10% tfnh theo cao dgc c h i l t n u d c t u d u g c lidu Kidn long ddm dat tidu chuan D u g c d i l n Vidt Nam IV do bp mfln D u g c - T n j d n g Cao dang Y te Hd Nfli bdo che, kem dgt tidu chuan ca s d .
T h u o c , hoa chat, may m o c p h ^ c vu n g h i d n cQ>u
- Ggc cat m i l n g hinh vudng dien tich 6 cm^
gac bdng, kinh Idp, t h u d c do dp day, t h i l t bj sinh t h i l t (biopsy punch) d u d n g kich 7 mm, can dien t i j (dfl chinh xac 0,001 g).
- Ddu croton (Hang san xuat: Sigma - D C F C ) , dung moi aceton, betamethason dang kem bdi 0,05% (Hang sdn xuat: PT. Tempo Scan Pacific Tbk - Indonesia).
D d n g vdt thiFC n g h i d m
Chugt nhat trang chCing Swiss, ca 2 glong, khoe manh, trpng lugng 25,0 ± 2,0 gam do Vidn V d sinh djch te Trung u a n g cung d p .
Tho chiing Newzeland White, ca 2 g i l n g , khde mgnh, trgng l u ^ n g 1 , 8 - 2,5 kg.
P h u a n g p h a p n g h i e n c u u Nghien ciru tinh an toan tren da Mfl hinh nghien cuu d u g c t h i l t k l vd t i l n hdnh dya trdn hudng ddn cua OECD vd Lidn minh Chdu Au '^'^. Thd d u g c boi 0,5 g c h i phdm nghidn ciJu tren didn tfch 6 cm^ (2,5 cm x 2,5 cm), phin da khflng boi t h u l c d u g c s u dung lam doi chdng:
Sau 4 gid bdi t h u l c va ta dugc, thao bd tat d bdng ggc ra khdi lung thd va n>a sgch thuoc hode td d u g c bang n u d c sgch. Ddnh gid va tinh d i l m cdc chJ so ve ban dd (erythema), phu n l (oedema)
TAP CHf DirOC HQC - 04/2013 (S6 444 NAM 53)
Nghidn ciru - Ky thuat
tgi thdi diem 1 gid, 24, 48, 72 gid sau khi logi bo thu6c. Niu cd tin thuang, theo ddi thd 14 ngdy d l ddnh gia kha ndng phyc hll. Khi tin thuang da hoi phgc thi ngung theo doi.
Tic dung ch6ng viem da cap tinh t§ii cho Nghien ci>u theo mfl hinh cda Tubaro'^l 50 chu$t nhdt trdng dugc chia ngdu nhidn thdnh 5 16, moi Id 10 con:
Ld 1 (Md hinh): Gdy mdhinh
L6 2 (Betamethason): Gdy mfl hinh + Boi betamethason
Lo 3 Thulc thd" ning d0 thlp: Gdy mfl hinh + Boi kem Viemda AD 5%
Ld 4 Thudc thi> ning dp cao: Gdy md hlnh + B6I kem Viemda AD 10%
Ld 5 (Td dugc): Gdy mfl hinh + Bdi td dugc 0 tat ca d c chuOt dugc bfli thulc thu vd gdy mfl hinh d tai phdi, tai trdi khdng tdc dgng gi. chuot dugc do chieu day tai d tit d d c 16.
Do chllu ddy tai tai vj trf sat dinh cua tai cdeh xa chdp sun vdnh tai. 6h sau khi gay md hinh, chugt dugc gilt, tai chugt dugc do Igi chilu day, sau do lay d phdn tmng tam vdi dudng kinh 7 mm bang dyng cu sinh thilt d l do can nang.
Xu I'f k i t qua
So ligu dugc nhdp hang phin mIm Excel 2007- Microsoft, xu ly bang phin mIm SPSS 16.0. So ligu dugc bieu dien dudi dang X ± SD.
Kilm djnh cdc gid tri bang test t - Student, test t ghep d p , test ANOVA 2 chieu.
Ket qud Tinh an todn tren da
Bdng 1: Kit qua thCe lilch uTig da cua cao d$c chiet niKrc tir ki§n long ddm
Th6Th6 1 Th6 2 Th6 3 Trung binh
1h CP
1 0 0 0,3
Ch 0 0 0 0
Ban do 24 h CP
1 0 0 0.3
Ch 0 0 0 0
48 h CP
0 0 0 0
Ch 0 0 0 0
72 h CP
0 0 0 0
Ch 0 0 0 0
1h CP
0 0 0 0
Ch 0 0 0 0
Phuni 24 h CP
0 0 0 0
Ch 0 0 0 0
48 h CP
0 0 0 0
Ch 0 0 0 0
72 h CP Ch
0 0 0 0 0 0 0 0
Cao dgc chilt nudc khdng gdy kich ung da
thd d tit d cdc thdi ^ I m nghidn cd'u. Vi sau 72 h khdng cd diu hieu ton thuang ndn thd khflng
Bdng 2: K4t qua thu" kich u-ng da cua cao d$c chiSt ethanol tu" kien long d&m tilp tyc dugc theo doi va khdng ldm gidi phau bdnh xdc djnh ton thuang.
Thd
Th61 Th6 2 Th6 3 Trung binh
1h CP Ch
1 0 1 1 0 0 0,7 0,3
Bandd 24 h CP
1 1 0 0,7
Ch 2 1 1 1,3
48 h CP
1 0 1 0,7
Ch 1 1 1 1,0
72 h CP
1 0 0 0,3
Ch 1 1 1 1,0
CP 0 0 0 0
1h Ch
0 0 0 0
Phi) 24 h CP
0 0 0 0
Ch 0 0 1 0,3
ne 48 h CP
0 0 0 0
Ch 0 0 0 0
72 h CP Ch
0 0 0 0 0 0 0 0
Da thd bfli cao ddc chllt ethanol, tgi thdi nh$ (tmng binh ban do la 0,7), tgi thdi diem 72h:
(Sim 1h, 24h, 48h: thulc gay kfch Crng mO-c dO khflng kfch ung (trung binh 0,3).
Bdng 3: Kit qua fhu kha ndng gSy kich (mg da cua kem Viemda AD 10%
7Ji6 Th61 Th6 2 Th6 3 Trung binh
1h CP
1 0 0 0,3
Ch 0 0 0 0
Bandd 24 h CP
1 0 0 0,3
Ch 0 0 0 0
48 h CP
0 1 0 0,3
Ch 0 1 0 0,3
72 h CP
0 0 0 0
Ch 0 0 0 0
CP 0 0 0 0
1 h Ch
0 0 0 0
PhO 24 h CP
0 0 0 0
Ch 0 0 0 0
ne 48 h CP
0 0 0 0
Ch 0 0 0 0
72 h CP Ch
0 0 0 0 0 0 0 0
Ghi chu: CP: Ch4 phim; Ch: ChOng
T^ CHf DlTgfC HQC - 04/2013 (S6 444 N A M S3)
Nghien CCFU - Ky thuat
Da thd bfli kem Viemda AD 10 %: Tgi thdi dilm 1h vd 24h, xuat hidn ban do, sau 72h khong thay cdn ban dd tren da. Dilm kich ung Id 0,3 (trong gidi hgn binh thudng).
Tdc dung ch6ng vidm da cap tinh tai cho ciJa kem boi da Viemda AD
Chieu day tai chugt
MiK dg sung huySt phii ne tai chugt quan sat bang mit thuxmg
Cd sy khdc biet ro rdt giOa tai phai va tai trdi.
Tuy nhien d c tai chugt bdi thulc thi> vd betamethason tai phai da giam ro r#t xung huyet, tay dd vd gian mach so vdi d c tai khflng bfli thuoc vd bfli td dugc.
Bang 4: Chiiu d^y tai ben phai cua chuot truvfc va sau khi gay mo hinh 6 h
L61L6 Mo hinh Chilu day j r u d c 19.40 ±2,12 (X±SD,|im) Sau6h 27,00±4,74'
1.6 2 Bethamethason
19,40 ±1,90 21,80 ±2,90"
1.0 3 Viemda AD 5%
20,20 ±1,99 22,00 ± 2,49'
L6 4 Viemda AD 10%
20,40 ±1,84 21,20±2,15*'
toS Taduvc 20,80 ±1,40 27,20 ±4,64*
ChO &tich: +: Khic biit so vai trutfc. test t ghep c$p, p<0,05
*; Khic bi§t so vai 16 md hinh tai cOng thai diem, test t-Student, p < 0,05 Lfl bdi betamethason chilu ddy tai d tang nhung khflng khac bigt so vdi topdc nghien ci>u. L6 bdi Viemda AD 5% va Viemda AD 10% diu dm Ld mfl hinh chi dugc bdi diu croton, chieu
day cua tai phal tdng len rd rgt, dilu dd cho thay tac dung gdy vidm d p cija dau croton tren tai
chugt nhat. thay sy gidm chilu day tai phal so vdi 16 mo hinh.
Khoi iwgng tai chugt, muv dg uv che vi§m Bang 5: Khdi Iwqmg tai chugt va mux: do u-c chi vi&m
Ldi Ld Afd hinh Khoi luwng tai Tr^i 13,6 ± 2,12
(X±SD,iig) PhSi 31,10±5,11' iyli>c do i>c che vi&m (%)
Ldi Bethamethason
13,50 ±1,58 19,50 ±2,84*'
65,7
1.6 3 Viemda AD 5%
11,20±1,69 25,20 + 2,53*"
20,0
Ld4 Viemda AD 10%
13,90 ±2,02 25,40 ± 2,37*"
34,3
LdS TadifOK 11,60 ±0,52 28,00 ± 2,27*
6,3
(y 16 md hlnh, khoi lugng tai phai tang rd rdt
so vdi tai ben trdi, dilu dd chirng td tdc dung gdy viem d p (phu nl) cua dau croton. Lfl bdi betamethason, khii lugng tai gidm ro rdt so vdi 16 mo hinh, muc do uc che viem cua betamethason Id 65,7 %. L6 b6i Viemda AD 5%
va Viemda AD 10% diu cho thay tac dung <rc chi viem rd rdt.
Ban ludn
v l tinh an todn cua kem bdi da Viemda AD tCr kien long dd'm
Danh gia kha nang gdy kich ung da la xet nghigm bat buOc doi vdi d c san pham thuflc va my phdm bfli ngodi da. Day la mgt khfa canh trong nghien cuu ve tinh an todn trdn da. Kit qud nghidn cCcu cho thay cao dgc chiet ethanol gdy kfch Ci-ng trdn da thd, cao dgc chiet nudc khdng gay kich ung. Do vdy cao dgc chilt nudc dugc lya chon sCr dung d l bao chi dgng kem
bdi da Viemda AD. Nghidn cCru tilp theo v l tfnh an toan tren da cho thay dang kem bdi da Viemda AD khdng gay kich ung da tren thd.
Ve tac di^ng chong viem tgi cho cua kem bdi da Viemda AD tir Itien long dd'm
Dau croton la san pham tO hat cua d y ba dau {Croton tiglium L.), thu§c hg Thau dau, mOt loai thyc vdt bdn dja vd hay dugc trong d An Dg. Khi dung ngoai da, dau croton gdy kfch dng manh. Hogt chat chinh cila diu croton Id chit hO'u ca dugc ggi la phorbol ''*'.
Ca che gay kich ung da cua ddu croton dugc giai thich thdng qua phosphollpase A2 (PLA2). Diu croton lam tang hogt tfnh ciJa PLA2 din tdi tang tgo acid arachidonic va sau dd Id cdc ylu t6 lidn quan tdi qud trinh vidm nhu' leucotrien vd prostaglandin. Ca che gay kfch Crng da cua diu croton cd sy tham gia hogt hda cDa hai hg enzym la cydooxygenase (COX) v|
T ^ CHi DlTQC HQC - 04/2013 (S6 444 N A M 5
0 Nghien ciju - Ky thuat
lipooxygenase (LOX). Mo hlnh ndy da d u g c n h i l u nhd nghien c u u tren t h i gidi sO- dyng d l ddnh gid tdc dgng chong vidm cua cdc t h u l c hode d u g c chat '^.
Betamethason (mOt logi corticoid) d u g c si>
dgng d l lam t h u l c d i i chCrng d u a n g . Corticoid dd d u g c chung minh Id co tac dgng mgnh n h i t tren m6 hinh vidm d p bang dau croton '^'. K i t qua nghidn c u u cho thay kem Vidm da A D 5% vd 10% cd tdc dgng chong viem d p tgi ch6 t i t , t h i hidn qua gidm mi>c d6 gidm dQ ddy tai vd trgng l u g n g tai so vdi lo md hinh, s y khac bidt cd y nghTa thong kd vdi p<0,05; tdc dyng t u a n g d u a n g v d i kem bfli betamethason 0,05%. Co t h i gial thfch k i t qua c h I n g vidm d p tgi ch6 cda kem Vidm da A D td do thdnh p h i n gentiopicrin trong d u g c lidu long d d m . Theo tdc gid Trung Quoc thdnh phan gentiopicrin trong long ddm cd tac dgng c h I n g vidm do (re c h i manh phosphollpase
Ket lu$n
- Kem bdi da Viemda AD 10% tinh theo cao ddc c h i l t nuo'c tir d u g c lidu kidn long ddm khdng gdy kich ung da trdn thd;
- Kem bdi da Viemda AD 10% tinh theo cao dgc c h i l t n u d c tu" d u g c ligu kidn long ddm cd tdc dgng c h I n g vidm d p tinh tgi ch6 trdn md hinh gdy vidm tai d p b i n g d i u croton.
S u m m a r y
Dermatic initance of the aqueous and ethanollc extracts from Gentiana rigescens Franch. on rabbits and topic acute anti- inflammatory activities of the aqueous extracts of Gentiana rigescens Franch. on mice were expenmentally evaluated, the former in temis of the level of edema and dermatitis , and the later
by the model of croton oil induced inflammation, comparing the decrease in the thickness and weight of the ears of the extract-treated mice with the controls. On rabbits, the aqueous extracts of G. rigescens exerted no Irritant effects to the skin; in contrary, the ethanol extracts did significantly. In mice, the water extracts of G. rigescens proved topic acute anti- inflammatory effects at both concentration of 5%
and 10 %.
Key words: Gentiana rigescens, demal irritation, anti-Inflammatory, effect.
Tai lieu tham khao
1. Ana Gallegos Saliner, Grace Patlewicz &
Andrew P. Worth (2007), "Review of Literature-Based Models for Skin and Eye Irntation and Corrosion", European Communities.
2. Organisation for Economic Cooperation and Development (2002), "Guideline for testing of chemicals:
/\cute Demal Irritation/Comasion", OECD 404.
3. A Tubaro, P. Ori, G. Delbello. C. ZiJIi, R. Delia Loggia (1985), The croton CA\ ear test revisited.
Agents and Actions. 17'(3-4) 347-349 4. McColl SR, Hurst NP & Cleland LG (1986),
"Modulaton by phortiol myristate acetate of arachidonic acid release and leukotriene syn-thesis by human polymorphonuclear leukocytes stimulated with /\23187", Biochemical and Biophysical Research Communications,^4•\•. 399-404.
5. Ezequiel Paulo Viriato, Erica Silva Bianchetti, Keldm Costa dos Santos et al (2009), "Study of high dilutions of copaiba oil on Inflammatory process", int.
J. High Dilution Res.. 8(26): 9-14
6. Wang Shi jun, Wang Zhi min (2009), "Nghidn CCFU thanh phdn hoa hpc va hoat tinh du'p'c ly lodi long ddm" - T^p chi Trung dirQC Trung Quoc. Vol.
34(23), p 2987-2994
7. Wang Xiang mei (2007), "Nghien cuu tac dyng dugc ly cua lodi long ddm". World Health Digest Medical Periodical, Vol. 4(11), p 21-22.
C a c h x a c d j n h . . . (Tiip theo trang 03) hon hode d i u nhd han ED50 thi khong t h i tfnh d u g c ED50, hogc tinh bang phdp ngoai suy thi k i t qud khdng chinh xdc.
Phuang phap kinh d i l n la phuang phdp t h u n h i t ( t i l n hdnh nhu xdc djnh LDSQ), nhung ton n h i l u thdi gian vd cdng si>c han, va k i t qua cung khflng chfnh xdc han phuang phdp ddng
giay ke logarit-probit, n i u chi mu6n xac c^nh ED50. N i u cd d i l u kign, cung nen xdc djnh thdm bdng phuang phdp Excel tren mdy vi tinh, r l i k i l m tra k i t qua cua 2 phuang phdp. Neu 2 phuang phap cho k i t qud xap xl nhau, d t h i tin t u d n g d k i t qud tinh todn. Neu k i t qud khdc nhau n h i l u , d n phai tinh toan lal.
