Nghien CCFU - Ky thuat
2. P C. Dandlya, S. K Kulkarni, (2005), "Introduction to pharmacology", Vallabh Prakashan New Delhi, pp.
247
3. J. H. Lawrie and M B. Glasg (1965), "The measurement of gastric acid", Postgrad. Med. J., 41, pp.408.
4 Mohammed A., Ravi Kumar J , Santosh H Y.
and Nagashmthi M. H.(2008), "Antiulcer activity of anisochilus carnosus leaf extracts in pylorus ligation rats", Indian Drugs 45 (12)' 979,
5. P. Muthusamy A. Jerad Suresh and G.
Balamumgan (2009), "Antiulcer activity of Azima tetracantha A biochemical study". Research J. Pharm.
and Tech. 2 (2), pp. 344-348
6. Dinesh Kumar Patidar (2011), "Anti-ulcer activity of aqueous extract of Murraya koenlgil In Albino rats",
International Journal of Pharma. and Bio. Sciences, 524-529.
7. Do Mmh Quang va CS (2000), "Tham do mgt so tac dung dugc 1^ cCia c h l pham Magaldrak (Malumix) tri viSm loet da day ta trang", T^p chi Dwac hoc, 285, trang 15'-17.
8. Padmaja Udaykumar (2005), "Textbook of medical Pharmacology" CSS Publishers, New Delhi, pp.317
9. Vien Dugc lieu (2006), "Phuang phap nghien cuu tac dung dugc ly cua thuoc tie duoc thao", Nha xuat ban Khoa hgc va Kythuat Ha Npl, trang 163-164 10. H. Gerhard Vogel (Ed.) (2002), "Drug discovery and evalution", Pharmacological assays, Spinger veriag Berlin Heidelberg New York, pp. 827 - 868.
{Ngiy nhin bil: 20/11/2014 - Ngay duyet dang; 01/10/2015)
vJDanh gia tac dung chong viem thyc nghiem cua cao dac bao che tir dich chiet nvo^c
bai thuoc EZ
Nguyin Manh Tuyfin'*, Phung Hoa Binh*, Le Thi Minh^
Nguyin Duy ThuanS Pham Thi Van Anh'
'Truang Dai hoc Duac Hd Noi,^Hoc vten YDuac co truyen VietNam
^Trucmg Dai hoc YHd Ngi 'E-mail: [email protected]
Summary
The viscous aqueous extracts from EZ remedy were evaluated for local anti-inflammatory effects by the croton oii ear test, acute anti-inflammatory on the carrageenin-lnduced edema In hind paws of mice, and chronic anti-inflammatory by granuloma test on mice at doses of3.5g and 10.5g/kg body weight of mice (corresponding to 1 and 3 times the human clinical doses, respectively). The extracts showed no exhibited no anti-inflammatory effects by the croton oil ear test, but acute and chronic anti-inflammatory effects through inhibiting carrageenin-induced edema in hind paws 4 and 6 hours after treatment, significantly decreasing granuloma weight, histological indexes as compared with the control (p<0.05).
Keywords: EZ remedy viscous extract, anti-inflammatory, experimental.
Dat van de
Phuong I h u l c EZ d u g c xay dung dua tren CO s d ly lugn y hoc c l truydn (YHCT) k i t hgp vdi cde k i t qua nghien cfiu Ihuc nghiem d l d i l u tri vidm da co dja (eczema). Trong bao cao trudc
day, cao dde d u g c bdo e h l tfi djch chief ethanol ciia bai thuoc da the hien tac dung chdng viem rd rgt Irdn fhuc nghidm M. Bai bao ndy trinh bay k i t qua nghien cfiu tde dyng c h i n g viem frdn thgc nghigm cua cao dac dugc bao c h l t u dich chilf n u d c bdi t h u l c vdi cdc myc tidu:
50 TAP CHi D l " "
• Nghien CCFU - Ky thuat
- Danh gia tic dung chdng viem cip tai cho tren chuot nhit tring.
- Danh gia tic dung chdng viem cap vi m^n tinh toan Uian tren chuot nhit tring.
Nguyen vat iieu, phuorng phap
nghien CIJUNguyen lieu nghien cunj
Bai thuoc EZ: gom (dm ngdn dang (Cauiis cum folium Lonicerae), nuc nac {Cortex Oroxyli), hoe hoa {Flos Styphnc^obii japonid imatun'), Uiuong nhT t u (FrucfusXantfiHsfrumara), hoang ba {Ck>rtex Pheltodendrf)... dat tieu chuan DDVN IV.
Cao d^c du-oc bao che tw djch chiet n v d v bii thuoc EZ (ggi tat la cao EZ-Hfi): Cdn ridng tung d u g c lieu da d u g c x u ly dat f leu chuan theo khoi lugng t u o n g ung vdi 5 thang (300 g d u g c ligu). C h i l l x u l l bdng nude theo phuong phdp sac, dun s6i 3 l l n , mdi lan 1 gid, lugng nude gap 6 lan d u g c lieu. Lgc ndng qua vai ggc, ly tam logi lap chat la lfing va ggp djch c h i l l . Cd dac thu d u g c 73,8 g cao dgc, hay 1 g cao dac tuang ung vdi 4,1 g d u g c lieu.
• Hda chit, thudc thw: Aspirin - Aspegic (DL- lysine Acetylsalicylale) gdi bgt 100 mg (Sanofi Aventis). Methyl prednisolon (Medrol) vidn nen 4 mg (Pfizer). Dung djch carragenin 1 % . Dau croton (Sigma - Due). Cdc hda c h i t khac dal lieu chuan phan lfch.
* Phuong tiin: Mdy do viem Plelhysmomefer No 7250 cua Hang Ugo-Basile (Italy). Thudc do dg day (do chfnh xac 0,02 mm), T h i l t bj sinh Ihilt (biopsy punch) d u d n g kfnh 7 mm, can dien l u (do chinh xac 0,001 g).
D o n g vat nghien efi'u
Chupt nhai trang chung Swiss, d 2 g i l n g , khoe mgnh, Irgng lugng 18 - 22 g do Vien Ve sinh djch te Trung u o n g cung d p .
P h u a n g phdp nghien cuoi
Dinh gii tic dung chong viem cap tai chd Chugt n h i t trdng d u g c chia n g i u nhidn thanh 416, moi 16 10 con nhu sau:
Ld 1 (md hinh): Gay mo hlnh + cho u l n g nude d t 0.2 ml/10 g chupt
Ld 2 (chung duong): Gay md hinh + cho u l n g
methylprednisolon 20 mg/kg chudi.
Lo 3 (cao E Z - H p lieu Ihap): Gay md hinh + cho u l n g cao E Z - H p l i l u 3,5 g/kg ehudt (luong fing 14,4 g d u g c lieu (DL)/kg).
Ld 4 (cao EZ-HjO l i l u cao): Gay md hinh + cho u l n g cao E Z - H p l i l u 10,5 g/kg ehudt (tuong fing 14,4 gDL/kg).
Chugl d u o c cho uong t h u l c t h u 2 ngay frudc khi lhfi nghiem. Ngay t h u 3, frudc khi gay md hinh 1 gid vdi lieu lugng nhu tren, ehudt duge gay m6 hinh b i n g cdch bdi 20 pl dung dich d i u croton (40 mg d i u pha trong 2 ml aceton) len mgi ngoai tai phai. Tai Irai khdng boi d l d l i e h i l u . Trudc khi gay md hinh, chupt d u g c do c h i l u ddy tai tgi vj tri sdt dinh d a tai each xa chdp sun vdnh tai.
Sau 6 gid, chugl dugc gdy md bang thiopental, do lgi e h i l u ddy tai chudi. Cat d phan laing tam vdi dudng kinh 7 mm bdng dyng cy sinh t h i l t can xac djnh k h l i iugng '^'^.
Dinh gii tic dung chdng viem cip toan than
Chugt n h i t trdng duge chia ngau nhien thdnh 4 16, mdi 16 10 con;
Ld 1 ( d l l chung): eho u l n g nudc d t 0,2 ml/10 g chugt
Ld 2 (chung duang): cho u l n g aspirin lieu 200 mg/kg chugt
Ld 3 (cao E Z - H p l i l u t h i p ) : cho u l n g cao E Z - H p l i l u 3,5 g/kg chugt
L6 4 (cao E Z - H p l i l u cao): eho u l n g cao EZ- H p lieu 10,5 g/kg chugt
Chugl d u g c u l n g I h u l c 3 ngay lien tyc Irudc khi gdy vidm. Ngay l h u 3, sau khi u l n g thuoc lhfi 1 gid, tiem canrageenin 1 % (tnang nudc m u l i sinh ly) 0,05 ml/chupt vao gan ban chdn sau, ben phai. Do the tfch chan ehudt (den khdp co chan) bdng may Plethysmometer d cac thdi diem:
trudc khi gay vidm (VO); sau khi gay vidm 2 gid (V2), 4 gid (V4), 6 gid (V6) ^-^K
Dinh gii tic dung chong viem man tinh toin thin
Chugt n h i l trdng d u g c chia ngdu nhien Ihdnh 4 Id, moi Id 10 eon:
T^P CHI DirpC H p c - 10/2015 (SO 474 NAM 55)
Nghien CIFU - Ky thuat
Lo 1 ( d l i chung): cho uong nudc d t 0,2 m l / l O g ehudt
Ld 2 (ehung duong): eho u l n g methyl prednisolon l i l u 10 mg/kg chugt
Lo 3 (cao E Z - H p l i l u Ihap): eho uong cao EZ- H p lieu 3,5 g/kg chugt.
L6 4 (cao EZ- H p l i l u eao): eho uong cao EZ- H p lieu 10,5 g/kg chugt.
Cay amianl trpng lugng 6 mg (sgi amianl cd dudng kinh khoang 2 mm d u g c vd trdn va s i y tigi k h u i n trong 1 gid d 120°C, t i m carrageenin 1%) vao da gay chudi. Sau khi d y amiant.
Bang 1 : Chiiu day tai chuot truoc
chuOt d u g e eho uong n u d c d t hode I h u l c thfi lien luc trong 10 ngay. Ngdy thu 11, g i l l chugt bdc tdch khoi u hgl, dem say kho d nhigt do 56°C Irong 18 gid. Can trpng lugng u hgt sau khi dd d u g c s i y khd f**-^.
X U I ly s o lieu: So lieu d u g e b i l u d i l n dudi dgng X ± SD, kiem djnh bdng test I - Student sg khdc bigt eo y nghTa thong kd khi p < 0,05.
K e t q u a n g h i e n CLPU
Tdc d u n g c h o n g viem cap tai c h o tren tai chugt nhat bang dau croton
vi sau khi giy viem 6 gia (n = 10)
1 Lo 1 (mo hinh) 2 Lo 2 (mettiyl prednisolon) 3 L6 3 (cao EZ-Hp Illu thap) 4 L64(caoEZ-H20lieucao)
0,02 3,5 10,5
Chieu day Tailr^i 21,2013,29 21,00 ±4,32 20,6014,01 19,80 + 3,05
lai chuot, ( X iSD, IDjjm) Talphai 27,00 +3,68*
23,40 + 3,12 25,40 + 3,53*
25,20+ 3,2r (Ghi chu: so Scinh trum sau. +, p < 0,05; so vdi nhdm chung: *, p < 0,05)
Khdng cd s g khac bigt v l chieu day lai ben phai d thdl d i l m Irudc nghidn cfiu gida cde Id.
0 16 md hinh, ehilu ddy ciia tai phdi fang ldn rd rgt so vdi trudc khi gay viem (p < 0,05). Ld u l n g methyl prednisolon, chieu day lai cd tang nhung khdng cd s g khac biet so vdi trudc khi gay viem (p
> 0,05). Nhung s g khac bigt ed y nghTa t h i n g kd
so vdi Id md hlnh (p < 0,05), I h l hign lac dyng c h i n g viem rd ret cua thuoc d l i chung duong.
Cao E Z - H p lieu 3,5 g/kg va l i l u 10,5 g/kg mdi chl t h i hidn cd xu hudng gidm dp day tai chugl so vdi Id md hinh, s u khac bidt khdng ed y nghTa t h i n g ke (p > 0,05).
Bang 2
Lo 1 (mo hinh) L6 2 (methyl prednisoion) L63(caoEZ-HjOli§uthap) Lo 4 (cao EZ-HjO iilu cao)
: Khoi luK^g tai chuot va miK
Lieu (g/ltg) —
0,02 3,5 10,5
m
LK ChS Viem (n =10)Ktiol Itfifng tai chugt, (X±SD, mg}
Tai fr^i 14,20 + 2.66 12.90 + 1.45 14.30 + 2.54 14.00 + 1.49
Taiph^i 33.80 + 6.37 17.60 + 3 . 3 f 29.30 + 3.06' 3 0 . 9 0 1 3 , 3 5 '
I^Ov do ipc die viem
(%)
44,08 12,72 8,88 (Ghi chu. so sanh trwdc sau: +, p < 0,05; so vdi nhdm chimg: '. p < 0,05)
(y Id md hinh, k h l i lugng tai phai tdng ro ret so vdi tai ben trdi. Ld uong methyl prednisolon, k h l i lugng tai giam rd ret so vdi Id md hinh (p < 0,05), mfic dp uc che viem cua methyl prednisolon la 44,08%.
Cao E Z - H p lieu 3,5 g/kg vd 10,5 g/kg d i u cd xu hudng giam khoi lugng tai chugt so vdi Id md hinh vdi mfic do u c e h l viem l l n lugt la 12,72%
va 8,88%. S g khdc bigt khong ed y nghTa thing kd (p > 0,05).
52 TAP C H i Dni'f*^ ^ " ' ^
• Nghien CIFU - Ky thuat
Tac d u n g chong viem cap toan than
Bang 3: Anh hwang cua cao EZ-Hp din do phii chin chugt (n = 10) Sau2gio'(VJ Sau4g'ia(VJ Sau 6 giir {VJ (9^9) DQphucy.) "^"^iH?" Do phu 1%) .. r. BQpt}u(%) , / , Lo 1 (dol chijng)
Lo 2 (aspihn) 57,47 ±19.02
p,^<0,05
Lo 3 (cao EZ-Hp lieu Uiip) 3,5 ^'^^ * g J J ^ -17,0: 66,67 + 18.38 p,_3<0,05
L64(caoEZ-H,Olieucao) 10.5 ^ ' ^ ^ ^ ' J * ^ -70.74 90.24 ±29,79 P M > 0 . 0 5
61,06 ±23,62 p,^>0,05 23,91 A s p i r i n l i e u 2 0 0 m g / k g c d t a c d y n g e h d n g v i e m
d p lgi tat ca cac thdi diem nghidn cfiu (p < 0,05).
Cao E Z - H p l i l u 3,5 g/kg ed tdc dung ldm giam phii chdn chugl d thdi diem 4 gid va 6 gid (P < 0,05).
Cao E Z - H p lieu 10,5 g/kg chi ed xu hudng gidm phu chan chugl d cac thai diem sau gay viem 4 gid va 6 gid ma khdng t h i hidn tae dyng chdng viem (p > 0,05).
Tac d u n g chong viem man toan than Bang 4: Anh huong cua cao EZ-Hp len khdi
Iwfjng u hat (n = 10) .. Ueu Kholluvmgu
(9*a) ( " 9 ) psovdi
161 psovdi
162 L61 (Sdi chimg) - 56.23118.67 Ld2(melhyipiednisoion) 0.01 43.3617.58 L63(caoEZ-H,Oiieuthlp) 3.5 40.45ll3.49 L64(caoEZ-H,Oiieucao) 10.5 42.8611474
<0.05
<0.05
<0.05
>0.05
>0.05 Methyl prednisolon lieu 10 mg/kg/ngay ed tae dung Idm gidm trgng lugng k h l i u hgt so vdi 16 chung (p < 0,05), the hien tdc dung c h i n g viem rd ret.
Cao E Z - H p d d 2 l i l u t h u d i u Idm glam Irgng lugng k h l i u hgl so vdi Id chung (p < 0,05), tdc dung ndy tuong duong vdi methyl prednisolon lieu 10 mg/kg ( p > 0,05).
Ban iuan
^ e tac dyng chong viem cap tai chS K i t qud nghidn cuu eho thiy, cd s g khac bidt rd rdt v l chieu ddy vd k h l i lugng tai ehudt d Id md hinh trudc vd sau gay viem 6 g i d , ehung Id md hinh d u g c Ihuc hign thdnh cdng. Thuoc doi
chfing duang methylprednisolon the hidn hidu Igc chong viem rd ret trdn md hinh nay, phii hgp vdi cae nghien cuu da cdng bo '^'.
S u khdc bigt vd dd ddy va khoi lugng lai ehudt d cdc Id l h u cao E Z - H p , l i l u 3,5 g/kg va 10,5 g/kg trudc vd sau gdy vidm d p b i n g tae nhan gdy viem Id dau cnaton khdng cd y nghTa t h i n g ke ( p > 0,05), Chung Id d lieu thfi vd trdn md hlnh t h u nghidm ndy, cao E Z - H p khdng ed lac dyng c h i n g vidm d p tgi chd, tuong l u cao dgc duge bdo e h l i d djeh c h i l t ethanol cua bai t h u l c d CLing muc l i l u t h u P'
Ve tic dung chdng viem cap toin thin Cao E Z - H p d l i l u 3,5 g/kg cd lac dyng giam do phu ban chan chugl (p < 0,05). Tac dyng ndy manh hon so vdi cao d u g c bao che l u djeh c h i l t ethanol d l i l u t h u l u o n g duong vdi ty le ue c h l 27,2% (cao nudc) > 7,68% (cao ethanol) vd 35,4% (cao nude) > 25,15% (cao ethanol) lgi thdi d i l m 4 gid va 6 gid. Tuy nhien, d l i l u thfi 10,5 g/
kg, thi Igi khdng t h i hien tac dyng, trai ngugc vdi xu hudng tac dung phy thugc vao lieu cua cao c h i l t ethanol trong l h u nghiem nay '^'.
Ve tic dung chdng viem man tinh toan thin
Cao E Z - H p , l i l u 3.5 g/kg va 10,5 g/kg d i u cd tdc dung giam trgng lugng u hat so vdi Id chung (p < 0,05). Mau nghien euu cd tae dyng chong viem man tinh tuang duong thudc doi chung duong methyl prednisolon l i l u 10 mg/kg chugt Cf l i l u 10,5 g/kg, cao E Z - H p cd tdc dyng chong vidm mgn linh lodn than tot han so vdi cao dac dugc bao c h l t u djch c h i l t n u d c cua bdi Ihuoc 18' va phil hgp vdi cae edng bd trudc day
TAP CHi DirpC H p c -10/2015 (SO 474 NAM 55) 53
• Nghien CI!FU - Ky thuat
v l tdc dung ching viem cua nhilu vj thulc trong bdi Ihulc nhu: don Id do, kim ngan dang, nuc ndc.'^'^i
Ket iuan
- Cao EZ-Hp khdng cd tde dyng ching vidm tai cho d lieu 3,5 g/kg vd 10,5 g/kg.
- Cao EZ-Hp cd tde dyng chong viem cap fodn thdn va chong d lieu 3,5 g/kg, khdng thay xu hudng phy thugc vdo lilu.
- Cao EZ-Hp cd fdc dyng chong viem man linh toan than d lieu 3,5 g/kg va 10,5 g/kg.
Tai iieu tham ithao 1. A. Tubaro, P Dri, G. Delbello, et al. (1985), "The croton oil ear test revisited". Agents and Actions, 17:
(3^) 347 - 349
2. McColl SR, Hurst Np Cleland LG (1986),
"Modulation by phorbol myristate acetate of arachidonic acid release and leukotriene syn-thesis by human polymorphonuclear leukocytes stimulated with A23187", Biochemical and Biophysical Research Communications,^4^: 399 - 404.
3. Orley, Vanesa Celeste, lldenize Bariiosa da Silva, et al. (2004), " Investigation of the anti- inflammatory and analgesic activities of a sample of Brazilian propolis". Acta Pharm. Bonaerense, 23 (3):
2 8 5 - 9 1 .
4. A.C.B. Paula, L.S.S. Hayash, J.C. Freifas (2003),
"Anti-inflammatory and antispasmodic activity of/pomoea imperati (Vahl) Griseb (Convolvulaceae), Farmacological effects of tpomoea imperati", Brazilian Joumal of Medical and Biological Research, 36:105-112
5. Ezequiel Paulo Viriato, Erica Silva Blanchetti, Kelem Costa dos Santos et al. (2009), "Study of high dilutions of copaiba oil on inflammatory process", InL J. High Dilution Res, 8(26): 9 -14
6. Vign Dugc li#u (2004), Cay thuoc va dong vat lam thudc & Vi$t Nam, tip 1, NXB Khoa hoc va Ky thu^t Ha Ngi, tr. 814 - 819, 930 - 934, 971 - 976,1044 -1047.
7. Vi§n Dugc ligu (2004), Cay thudc va ddng v$t tam thuoc d Vi$t Nam, tip 2, NXB Khoa hoc va Ky thuat. Ha NOi, tr. 106 -110, 480 - 484.
8. Nguyin Mgnh Tuyen, Phung Hoa Binh, LeThj Minh va cs (2015), "Binh gi^ tac dung chdng vi4m thu'c nghidm cOa cao dSc bao chi tCrdich chiet ethanol bai thuoc EZ", Tap chi Dwac hoc, 471 (55): 6-10.
{Ngay nhin bii; 04/08/2015 - Ngay duyit ding; 01/10/2015)
\(Phan tich dong thoi acid ferulic, scopoletin va xanthotoxin trong dirong qui di thirc bang
sac l^y long hieu nang cao l^et noi detector huynh quang (HPLC-FLD)
Dirorng Thu Hirong', Nguyen Thi Ha Ly^ Phvonig Thipn Thirong^*
'Hoc vien Qudn Y; ^Khoa Hoa phdn tich - Tieu chudn, Vien Duac lieu 'E-mail: [email protected]
Summary
An HPLC procedure was established for simultaneous determination of ferulic acid (AF), scopoletit) (SC) and xanthotoxin (XA) in Radix Angelicae acutilobae by an RP-HPLC-FLD system. Mobile phase - a stepwise gradient acetonltrile-methanol-water mixture; Flow rate - 0.5 ml/min; Detector - FLD (A^'=340 nm and ^^^=425 nm). The LODs were 1.00; 0.09 and 0.9 ng/ml, and the average of recovenes were 95.0%, 95.2% and 95.3% forAF, SC and XA, respectively The method proved practically helful for the quality evaluation and control of Radix Angelicae acutilobae and related materials, preparations.
Keywords: Angelica acutlloba, scopoletin, xanthotoxin, acid ferulic, HPLC-FLD.
Li i-rt/^ ur»(- _ 1 A/Tni ^ reiS Aid N A M ^Vt
