• Nghidn ciru • Ky thuat
Tai lieu tham khao 1. Bo Y td (2009), "ChuySn luan v l enzym chy- motrypsin (tr 157- 159) va c^c phu luc 9 6, 13 2. 13.6.
15 1 (tr PL 172-183, 250-310)", Duvn: diin Viet Nam 4. XB lan thu 4, NXB Y hpc, Ha Ndi.
2. Dd Dire Giao (1999), "Thuat toan - Phuang phap d | quy - Quan h$ - 06 thj hij-u hgn - Ldgic toan"
, Toan r& r^c, Nxb Ogi hpc Qudc gia H^ Npi, tr 5-127, 332-415
3. Layne. E (2008), "Spectrophotometric and turbidimetnc methods for measuring proteins' Methods in Enzymology 10, p. 447-455
4 Lauwers,A,Scharpe, 8 {^997), Pharmaceutical Enzyms, drugs and pharmaceutical sciences . Volume 84, Marcel Dekker, Inc , New York-Basel-Hong Kong.
5. Vincent Guyonnet, Franciszek Tluscik, Peter LLong, Antoni Polanowski, James Travis (1999), 'Purification and partial characterization of the pancreatic proteolytic enzyms trypsin, chymotrypsin and elastase from the chicken". Journal of Chromatography A 852 p 217-225
6 United States Pharmacopeial Convention, USP 30 - NF 25 (2008), "Prescnption and over-the-counter medicines". United States Pharmacopeia, p 1747 - 1748, p.2856 -2857 NY USA
{Ngdy nhan bdi: 16/03/2015 - Ngdy duydt ddng: 04/05/2015)
/Nghien cwu tac dung chong viem cap, giam ho va long dom cua cao long Vi ngan
tren thyc nghiem
Nguygn Thi Thu TrangS Bui Hong Cu-ong', Pham Thi Van Anh^
'Tntcmg Dai hoc Duoc Ha N$i
^Truang Dai hoc Y Ha Npi
*E-mail. [email protected]
Summary
Liquid extracts of the compound herbal remedy "Vi Ngan" (VE) were investigated for antitussive and expectorant effects aldoses of 63.36 g and 190.08 g per kg body weight of mice, and acute anti- inflammatory effects at doses of 42.24 and 126.72 g per kg body weight of rats. VE at dose of 126.72 g/kg b.w. significantly proved acute anti-inflammatory effects as compared with the control group. VE in dose 190.08 g/kg showed significant antitussive and expectorant effects.
Keywords: VE, anti-inflammatory, antitussive, expectorant.
Dat vdn dg
Cao long VT ngdn dyge didu ehd tir 12 v\
dyge li^u gdm: Kim ngdn hoa, xg can, bgc ha, hung ehanh, nuc ndc, cam thdo, cdt cdnh, bang dgi hdi. huydn sdm, mach mfln, thien mfln vd sinh dja. Co sfl' phdi hgp eae dygc lieu cua bdi thude dya tren kinh nghiem sy dyng thuoc eua Y hge cd tmydn vd tde dyng dygc ly da dygc nghien cyu cua cde dygc liflu'^''', nhdm hyong sii' dyng bdi thudc trong benh viflm hgng cap ttnh. Oe danh gid tdc dyng dygc ly eo lien quan ddn triflu
chirng trong benh ly viem hgng cap, nghien ciru ndy dygc tien hanh nhdm mue tiflu: Nghidn cOv tdc diing chdng vidm cip tren chudt cdng tring, tdc dung gidm ho, long d&m trdn chudt nhit trSng cua cao Idng Vfngdn.
Ddl tu'p'ng va phu'O'ng phap nghien CLPU
Chdt lieu nghien cu>u
Cao long Vi ngan: Kim ngan hoa, xa can
chidt bdng ethanol 70 % (ngam kiet), cat thu hdi
Nghien CIFU - Ky thuat
dung mfli; cdc vj cfln lai nau v ^ i nyoc, co djeh ehidt. Ggp 2 eao, co ddn eao long 5/1.
Cac hoa chat: Aspirin - Aspegic (DL-lysin Acetyl salieylat) gfli bflt 100 mg cua Hang Sanofi Arventis, Phap; Codein phosphat, N-acetyleystein (bigt d y g e Mueomyst gfli 200 mg), phenol dfl dang bgt tinh khidt (Hdng Sigma), n y o c mudi sinh ly, dung djch earrageenin 1 % va ede hfla chat khdc nhy formaldehyd, ... du tieu chuan phflng thi nghigm Tryong Dai hge Y Hd Ngi.
Dyng c(/ nghien cuv: Dyng cy do thd tich ehdn chuflt Plethysmometer No7250 eua Hang Ugo-Basile (Italy). Kit djnh lygng protein ciia Hang Hospitex Diagnostics (Italy). Dung djch xet nghigm mdu ABX Minidil L Blood Counter.
Ogng v|it thi nghigm
Chuflt nhat trdng chung Svwss, cd 2 gidng, khod manh, trgng lygng 18 - 22 g do Vifln Vfl sinh djch td Tmng y o n g cung edp. Chugt cong trdng chung Wstar, ea 2 gidng, khoe manh, trgng lygng 180 - 220 g eua Hpc vign Qudn y.
Phyong phdp nghien cu>u Nghien cuv tac dyng ching viem cip Md hinh gdy phii chdn ehugt bang carrageenini^ "1
Chugt edng trdng d y g c chia ngdu nhifln thdnh 4 Ifl, mdi Ifl 10 con.
- Lo 1 (ehirng): udng nyo'c eat lieu 1 ml/IOOg.
- Lfl 2 (ddi ehieu): uong aspinn lieu 200 mg/kg.
- Lfl 3: udng cao long VT ngdn lieu t y o n g d y o n g 42,24 g du'gc liflu/kg (Lieu gap 3 ldn lieu d y kien trfln Idm sdng).
- Lo 4: udng cao Iflng VT ngdn lieu t y o n g du'ong 126,72 g d y g c lieu/kg (Lieu gap 9 lan lieu d y kidn trfln Idm sang).
Chugt d y g e uong thude 4 ngay lien tyc t r y o e khi gdy viflm. Ngdy thir 4, sau khi udng thuoc 1 gig, gay viem bang each tiflm earrageenin 1 % (pha trong n y o c mudi sinh ly) 0,05 ml/chugt vdo gan bdn chdn sau, ben phai eiia ehuflt. Do the tieh chdn ehugt (den kho'p cd ehdn) bdng dyng eu ehuyen biet vao cdc tho'i diem- t r y o c khi gdy viem (VO); sau khi gdy viem 2 gifl' (V2), 4 gio (V4) vd 6 gio' (V6). Ket qua d y o c tinh theo cflng t h y c eua Fontainei^i.
M6 hinh gay viflm mdng bung bSng carrageenir^^^
Chuflt cong trdng d y g c bd tri thi nghiflm nhy tren. Chuflt d y g c udng thude hogc dung moi trong 4 ngdy lien tye toro'c khi gdy viflm. Ngdy thii' 4, sau khi udng thuoc 1 gio', gay viem mdng bung bang dung djch earrageenin 0,05 g pha vol fonmaldehyd 1,4 ml, pha trong nyfl'c mudi sinh ly 0,9 % vija dii 100 ml. Tiem vdo khoang mdng bung v&\ the tieh 2 ml/ chugt. Sau 24 g i o gdy viem, md d byng ehugt hut d]ch ri viflm. Do thd tich vd dem sd l y o n g bach eau/ml djch ri viem, djnh lyong protein trong djeh n viem, tinh so lygng bgch cau vd hdm lygng protein trong d djeh ri viflm eua tirng ehuflt.
Nghidn cipu tac dung giam hc/^i 40 chuflt nhdt trdng d y g c ehia ngau nhifln thdnh 4 Ifl, m6i 16 10 eon
- Lfl 1 (chu'ng): udng n y d e cdt, the tieh 0,2 ml/IOg.
- Lfl 2 (ddi chieu): uong eodein phosphat lieu 20 mg/kg.
- Lfl 3: udng eao long VT ngdn lieu tyong d y o n g 63,36 g d y g e lieu/kg.
- Lfl 4: uong eao long VT ngdn lieu tyong d y o n g 190,08 g d y g e lieu/kg
Chuflt d y g c uong n y o e hoac thudc thir lien lyc trong 3 ngdy vao cde budi sdng. Ngay thir 3 sau khi udng thuoc thii' 1 gio, tidn hdnh gay ho eho ea 4 Ifl ehugt bdng khi amoniae lidu 0,5 ml/
binh thuy tinh ehuyen dyng. Xde djnh thoi gian tiem tang (t) Id thb'i gian tinh tir khi thd chugt vdo binh den khi ehuflt xuat hign eon ho ddu tifln va tdng sd con ho tmng binh trong 10 phiit, so sdnh mire do tang (t) vd mire do giam s6 con ho cua cdc ifl thir so voi 15 chyng vd Ifl ddi chidu.
Nghien cuv tac dyng long tfflmra 40 chuflt nhdt trdng d y g c chia ngau nhifln thdnh 4 Ifl, moi Ifl 10 eon.
- Lo 1 (ehyng)- udng nu'oc cdt vfl'i the tich 20 ml/kg/ngdy.
- Lfl 2 (ddi chidu): N-acetyteystein 120 mg/
kg (can 180 mg pha voi nu'oc edt vira dii 30 ml, udng 0,2 ml/IOg).
- Lfl 3: uong eao Iflng Vi ngdn lidu t y o n g d y o n g 63,36 g d y g e ligu/kg.
- Lfl 4; udng cao Iflng VT ngdn lidu t y o n g d y o n g 190,08 g du'gc ligu/kg.
TAP CHI Diroc HOC - 5/2015 (S6 469 NAM 55)
• Nghien CIFU - Ky thuat
Cho chuflt udng thudc thir vol lidu nhy trfln trong 3 ngdy. Ngdy thy 3 (ngdy cudi eung) sau udng thudc 1 gio, tiflm mdng byng eho tat ea ede chuflt dung djch phenol do 0,25 % vfl'i lidu 25 ml/
kg. Sau 30 phut tiem nhde lai mflt lidu nhy vdy Chfl' 30 phiit sau, giet ehugt bdng gdy ngat hoi ehioroform, bflc lfl khi qudn va ludn vdo dfl mgt kim tu. Rira khi quan bang cdch diing bom tiem lay 0,5 ml dung djeh NaHCOj 5 % bom vdo khi qudn, hiit nhg nhdng rdi chuyen vao trong mot dng nghigm da dyge danh ddu tir tryoe.
Lam nhy vay 3 ldn ddi vfl'i moi con, tap tmng djch rira eua eimg mflt con trong mgt ong nghiem (the tieh dieh ara thu dygc cita ede chugt bdng nhau). Ly tdm djeh ru'a Idy djeh trong Do do hap thy quang (ABS) eiia djeh rya khi quan o' byoe sflng 265 nm.
K§t qua nghien CLPU
Tac dung chdng viflm cdp ciia cao long VT ngan Ket qua nghien ciru tac dyng ehdng viflm cap eiia eao long VT ngdn dyge ghi trong bdng 1 va 2.
Bang 1: Anh hu^ng cua cao Idng Vfngdn Idn khd ndng uc chi phCi chdn chudt
Sau 2 giir Sau 4 giir Sau 6 giir
Khananguc Khi nang 6c ^ Khi nang uo^
che phu so vol T ^ p ™ ^ P"^ sova ' Z cp ™ ^ P"^ ^° ""^
diCmg {%) ~' Ddphu(%)
±SE chiing (%)
Lo 1 {nir&c d t 1 ml/100 g) 25,34 ± 3,22 Lo 2 (aspinn 200 mg/kg) 9.30 + 1,15 Pj.,<0.001
23,82 ±2,85 P.,<0,01
30,25 ± 3,43 p,,<0,05 L6 3 (cao long VT ngSn tu-ang
dtfpng 42,24 g du-pc li#u/kg) 24,50 + 2,92
P 3.1^0.05
40.68 ±3,55 P 3.1^0.05
38,93 + 3,74 p , >0,05 Lo 4 (cao long VT ngSn tu'ong 22,25 ± 2,50
(Ju-cng 126,72 g du'oc lieu/kg) p (.,>0,05
40,09+4,76 P ^,="005
34,18 + 3,42 P 4-1^0.05 17,50
Nhdn xdt:
Aspirin lieu 200 mg/kg efl tac dung lam gidm dg tang thd tieh ehdn ehuflt fl'tat ca cdc tho'i diem nghifln ciru so vfl'i 16 chirng (p < 0,05).
Cao Iflng VT ngdn o' ea 2 lieu: lieu tyong Bang 2: Anh hi^dmg cua cao ldng Vingdn Idn thd
protein trong
dyong 42,24 g dygc lieu/kg vd 126,72 g dygc ligu/kg cfl tdc dyng Idm giam dg tdng the tich chan ehuflt sau khi gdy viflm fl' tat ea cdc thoi diem nghien ciru so voi Ifl chirng. Tuy nhien, sy khae biflt khong efl y nghTa thdng ke (p > 0,05).
tich dich ri viem, sfl limng bach cdu vd hdm limng 6 djch ri vidm
The tich dich ri viem (ml/10l}g chugt)
6 Itrgmg b§ch ciu Oong 6 djch n Ham lif^g prolan trong viem (G) 6 djch ri viem (mg) L61 (Chii-ng)
16 2 (Aspinn 200 mg/kg/ng^y) 1,23 + 0,29 p „ < 0 , 0 5
0,03+0,01 P M •= 0.01
0,01 + 0,003 p „ < 0,05 Lfi 3 (Cao V i ngan tuang duong
42,24 g duoc li#u/kg)
1,90 ± 0 , 3 1 Pj^,>0,05
0,05+0,01 p, ,> 0,05
0,02 + 0,002 Pj.,> 0.05 L64 (Cao VT ngan tirong dirong
126.72 g diro'c l!9u/kg)
1,48 ±0,07 P „ < 0,05 p^j> 0,05
0,04±0,002 P^i<0.01 P«>0.05
0,01 ± 0,001 p^,<0,05 P « > 0 , 0 5
Nhdn xdt:
Asprin lieu 200 mg/kg ldm giam ro rflt the tieh djch ri vidm, sd lygng bgeh eau vd hdm lyong protein so vfl'i 16 ehirng (p<0,05)
Cao long VT ngan o' cd 2 lieu (tyong dyong
42,24 g dygc lieu/kg vd 126,72 g dyge liflu/kg)
lam giam thd tich djch ri viflm, sd lygng bach cau
vd hdm lygng protein so vfl'i Ifl ehirng. Nhyng sy
Nghien CLPU - Ky thuat
khdc biet ehi efl y nghTa thdng ke o' lieu tu'ong Tdc d u n g gidm ho cua cao l o n g VT ngdn d y o n g 126,72 g d y g c lieu/kg (p<0,05). (Xem bdng 3, 4)
Bang 3: Anh hu^g cua cao ldng Vl ngdn Idn thdi gian tidm tdng xudt hi$n can ho Lo chugt
L61 (Chung)
Lo 2 (Codein phosphat 20 mg^g) Lo 3 (Cao long VT ngan biang duong 63,36 g duoc lieu/kg) Ld 4 (Cao long Vi ngan ttrong duong 190,08 g duoc lieu/kg)
n 9 9 10 9
Thai gian Him tang($)
±SE 118,00±22,05 226.00 ±44,43 240.40±59,19 268,89 ±59,45
Afiic dd tang thai gian tiim tang so voi chiing (%)
91,53 103,39 127,11
p (so voi chCm^
pj.,<0,05 p3.i>0,05 p^,<0,05 p^j>0,05 Nhdn xdt:
Codein phosphat lieu 2 0 mg/kg cfl t d c d u n g keo d d l tho'i gian tidm tang xuat hifln e o n ho tren ehugt nhat trdng so vo'i lo c h i r n g (p < 0,05), v&i m i r c tang la 9 1 , 5 3 %.
Cao Iflng VT ngdn fl' ea 2 lieu tu'ong du'ong
Bdng 4: Anh hirdng cua cao ldng Vingdn Idn tdng si con ho
63,36 g d y g c liflu/kg vd 190,08 g d y g e liflu/kg efl tae dyng keo ddi tho'i gian tidm tdng xuat hifln eon ho tren chugt nhdt trdng so vfl'i 16 chirng vfl'i mirc tdng Idn l y g t Id 103,39 % vd 127,11 %.
Nhyng s y khde biel eo y nghTa thdng ke fl' lidu t y o n g d y o n g 190,08 g d y g c li^u/kg (p< 0,05).
Lo L61 (ChCrng)
L6 2 (Codein phosphat 20 mg/kg) L6 3 (Cao long VT ngan tuong duang 63,36 g du-oc ti^u/kg) L6 4 (Cao long VI ngan tuong duang 190,08 g du'O'c liSu/kg)
n 9 9 10
9
T6ng so con ho trong
±SE 30,33 ± 6,30
9,33± 3,75 14,30 ±3,39
7,00± 3,34
10 phut MCPC do so v6
giam so con ho 16 chimg (%)
69,24 52,85
76,92
P (so vdi chiing)
Pj^< 0,05 P3 ,< 0,05 Pj.2> 005 p,.,< 0,001 p „ > 0,05 P M^ 0.05 Nhdn xdt:
Codein phosphat lieu 20 mg/kg cfl tdc dyng Idm giam sd con ho tren ehugt nhdt trang so vol lo chyng (p < 0,05), vol mu'c giam Id 69,24 %.
Cao long VT ngan lidu t y o n g d y o n g 63,36 g d y g e liflu/kg co tde dung Idm giam sd con ho o'
chuflt nhdt trdng so vfl'i Ifl chyng (p < 0,05), vfli mire gidm Id 52,85 %.
Cao long VT ngdn lidu t y o n g d o o n g 190,08 g d y g c lieu/kg eo tae dung lam gidm r5 rflt sd eon ho fl' ehuflt nhdt trdng so vfl'i 16 ehirng (p = 0,005), vfl'i m y e gidm la 76,92 %.
Tac d u n g long d o m cua cao long VT n g a n (Bang 5)
Bang 5: Anh hir&ng cua cao Idng Vfngdn ldn mdt dd quang Lo
L61 (Chimg)
Lo 2 (N-acetyl cystein lieu 120 mg/kg) L6 3 (Cao long VT ngan tuong duong 63,36 g diroc lieu/kg)
Lo 4 (Cao long VT ngan tifong (Jifong 190,08 g diioc lieu/Kg)
n 10 10 10 10
Mat d6 quang tSE 0,406 ± 0,036 0,528 ±0,023 0.44710,036 0,664 ±0,015
Uifc tang mat 06 quang so voi chimg (%)
30,05 10,10 64,28
p{sov6ichimg)
p^,< 0,05 p,.,> 0,05 p „ < 0,001 p „ < 0,001
TAP CHi Diroc HOC - 5/2015 (S6 469 NAM 55)
• Nghien CLFU - Ky thuat
Nhdn xdt
N - acetyl cystein lieu 120 mg/kg co tde dung lam tdng mdt do quang trong djeh ru'a khi quan chuflt nhdt trang so vfl'i 16 ehirng (p < 0,05), vol mye tang 30,05 %.
Cao Iflng VT ngdn lidu t y o n g du-ong 63,36 g dyge ligu/kg cfl tdc dyng Idm tdng mdt do quang trong djch ru'a khi quan chuflt nhat trdng so vol lo chirng, voi m y c tang 10,10 %. Nhyng s y khde biflt khflng cfl y nghTa thdng ke (p > 0,05).
Cao Iflng VT ngdn lieu t y o n g d y o n g 190,08 g d y g c li§u/kg efl tde dyng lam tang ro rgt mgt dp quang trong djeh rya khi quan ehuflt nhat trang so vfl'i Ifl chirng (p < 0,001), voi m y c tdng 64,28 %. MO'C tang ndy manh hon so v ^ i N - acetyleystein lieu 120 mg/kg (p < 0,001).
Ban luan v d bdi t h u d c VT n g a n :
Dya tren eong dung eiia eae vj thudc, lien he giii'a Y hgc cd truydn vd Y hge hifln dai, theo hu-gng ket hgp cdc vj thudc, nhdm gidi quydt cae tri?u chirng cua bflnh viflm hgng nhir: tae dyng khdng khuan (bge hd, hung chanh, kim ngdn hoa, xa can, thifln mfln, maeh mfln, eat ednh), chong viem (kim ngdn hoa, xa can, cam thdo), ehong di irng, tdng sire dd khdng (kim ngdn hoa, niic ndc), giam ho, long dom, sinh tdn dieh (cat cdnh, cam thdo, mgch mfln, thifln mfln)...i'-^i
Dya trfln phyong phdp bien ehyng lugn tn eua Dong y: Viem hgng (ehyng hdu ty) vol bieu hign su-ng, nflng, dfl, dau Id tinh trgng do khi nhiflt, dom kdt 6' phe vi, hode do ngogi ta xdm nhdp, nhidm khi dgc d|ch le, nhiflt dflc phe vj xong Ifln.
Men edn thanh nhiet giai dgc, sdt khuan, nhy xa can, kim ngdn hoa, nuc ndc, cam thao, ngodi ra CO bdng dgi hdi, huydn sdm...Ket hgp vfl'i ede vj thudc efl tde dyng thanh phe, hfla dam, ehf khdi dehd trg them tac dyng nhy eat ednh, thien mfln, mgeh mfln. cam thao...
v d cdc tdc d u n g : c h f l n g viem cdp, giam ho vd long d o m :
Trfln mo hinh gdy phii ehdn chuflt edng, khdng nguyfln sir dyng Id earrageenin, eo ban chdt Id polysaceharid gdn giong vol edu triic vfl vi khudn, vi vdy ddp irng midn djch cua eo the chu ydu Id ddp irng mien djch khflng ddc hi^u vfl'i s y
tham gia ciia ehii yeu la dai thye bao, bgeh cau mill trung tinh'^ ^l Bleu hien eiia qud trinh viem ndy Id gidn mgeh, baeh eau xuyen maeh, tdng tiet cdc chdt trung gian hod hge nhy prostaglandin, hjstamin, leucotrien, bidu hien quan sat thdy ehu ydu Id triflu chirng phii. Cao Iflng VT ngan o' cd 2 lieu: t y o n g d y o n g 42,24 g d y g e lieu/kg vd 126,72 g d y g e lieu/kg, ehya Idm Idm giam do tdng thd tich chdn chugt sau khi gay viem fl'tat ea eae thoi diem nghifln eiru so voi Ifl chirng.
Cao long VT ngdn lidu t y o n g d y o n g 126,72 g d y g e ligu /kg/ngay the hign tdc dung chdng viflm cap trfln chugt cdng trdng thflng qua Idm gidm ro ret thd tfeh djeh ri viem, sd lygng baeh eau vd sd lygng protein trong djeh ri viflm trfln mfl hinh gdy viflm mdng bung ehuflt bdng earrageenin. Tde dyng ndy t y o n g d y o n g vol tae dung ciia aspirin lieu 200 mg/kg.
Codein co tae dyng gidm ho do u'c chd tryc tidp trung tam ho, eodein d y g e chgn Idm thudc doi chirng tren mfl hinh gdy ho bdng khi amoniae 0'ehuflt nhdt trdng.
Cde kdt qud nghien ciru eho thdy cao long VT ngdn lieu t y o n g dyong 190,08 g d y g c liflu/kg efl tdc dyng Idm tang thdi gian tiem tang xuat hten eon ho vd Idm gidm sd eon ho eiia chuflt nhat trdng trfln mfl hinh gay ho bdng amoniae. Tdc dyng ndy t y o n g dyong vol tae dyng eua codein phosphat lieu 20 mg/kg.
Cao long VT ngdn 6' mu'c lieu t y o n g d y o n g 190,08 g d y g c liflu/kg efl tde dyng long d&m thflng qua Idm tang nong dg phenol dfl trong djeh rira khi phd quan chugt nhdt trdng. Tde dyng ndy manh hon tde dung cua N - acetyleystein lieu 120 mg/kg
Nhy vgy, cao Iflng VT ngdn lieu t y o n g d y o n g 190,08 g d y g e lieu/kg eo tde dyng gidm ho vd long dom tren chuflt nhdt trdng, tac dung nay phii hgp vo'i tdc dyng eOa ede vj thudc trong phyong thudc.
Kk luan
Cao long VT ngdn efl tdc dyng ehdng viflm edp trfln mfl hinh gay viem mdng bung fl' lidu t y o n g d y o n g 126,72 g d y g c lieu /kg/ngay, cfl tac dyng giam ho tren mo hinh gay ho bdng khi ammoniac
& lieu t y o n g d y o n g 190,08 g d y g e ligu/kg/ngay va efl tde dung long 6&m tren ehuflt nhdt trang a lieu t y o n g d y o n g 190,08 g d y g e lieu/kg/ngdy.
