Tki li^u tham khio
1. Nguyen Viet Than (2006), "Nghien ciiu d4nh gid thyc tr^ng chit lu-gng dugc li^u trgn thj tnr&ng Vift Nam hi?n nay". Mo c4o kSt qud nghign cuu <tk tdi cSp Bp Y t^. 2. Nguyin Kim Bich (2006), "PhSn tich vd xdc djnh cdc d?ic diSm h6a hpc d§c trung ciia duac ligu phyc vy c6ng tdc tigu chu&i h6a", bdo cdo kit qud nghign clini ^ tdi cap B6 Y t l 3. Du?c diln Vi^t Nam IV (2009), NXB Y hpc. 4. Pharmacopoeiae of The People's Republic of China (2010). volume 1. 5. Bp Y tl (2007), Tdi lieu h^i nghi dugc ligu todn quoc ISn thd hai, tr.100-105. 6.
Nguyen Thj Bfch Thu (2010), "Nghien cihi img dyng s^c k^ khi khii phi dl phSn tich du lugng m0t sl hod chdt bdp vg thuc v4t thu&ng diing" (2008-2009), bdo cdo kit qud nghign cihj dl tdi ckp Bp Y tl.
Tap chiDteac liiu, tap 18, so 1/2013 (Trang 14 - 20)
NGHIEN CU'U DOC TINH CAP, TAC DUNG BAO VE GAN VA TAC DVNG G A Y D O C T E B A O UNG THU' CUA XAO TAM PHAN
Nguyin Minh Kh&i, Phqm Thi Nguyet Hing, Bd Thf Phumg Vien Duac lieu
(Nhan bai ngay 22 thdng 01 nam 2013) Summary
Study on Acute Toxicity, Hepatoprotective Activity and Cytotoxic Activity of Paramignya trimera (Oliv.) Guillaum
Paramignya trimera (Oliv.) Guillaum has been knovm as the medicinal plant with maverllous treatment in Khanh Hoa provine, Vietnam, recently. However, there aren't any scientific evidences of chemical composition as well as toxicity and pharmacological effects. The aims of this study are to evaluate the acute toxici^, hepatoprotective activity on paracetamol-induced hepatic damage in mice and cytotoxic activity in vitro. The results show that LDo of methanol extract from Paramignya trimera (Oliv.) Guillaum is lower or equal 329.0 g/kg body weight for single dose oral administration. Oral administrations of methanol extract at dose 10 g/kg and 20 g/kg didn't show any changes in alanine aminotransaminase (ALT), aspartate aminotransferase (AST) or bilirubin on the model of paracetamol-induced hepatic damage in mice. Methanol extract, n-hexan fraction and the compound 6 (3,7-dimethyI-2,6-octadienyl)7-hydroxy-2H-lbenzopyran-2-one (1) showed cytotoxic activity in vitro against 5 cancer cell lines, in which n-hexan fraction and (1) exhibited moderate cytotoxic effects on human hepatocellular carcinoma (Hep-G2) and human epithelial cervical carcinoma Hela cell lines with ICjo values of 39.61 Hg/ml and 18.4 (iM, respectively. However, butaiol fraction exhibited no toxicity to 5 cancer cell lines in this study. Based on these results, it is necessary to tlirther study to evaluate the safety and pharmacological effects. ^^
Keywords: Paramignya trimera, acute toxicity, hepatoprotective activity, paracetamol induced hepatic damage, cancer cell lines.
1. D^t v^n de nhilu tin ddn cho rdng xdo tam phdn la thk' Cay xdo tam phdn cd tfin khoa hgc Id dugc, cd thfi chiia dugc nhilu. bpnh hilm Paramignya trimera (Oliv.) Guillaum, thudc ngheo. Tuy nhifin, chua co mgt nghien cftu hp Cam (Rutaceae). Theo tdc gid Pham nao trong vd ngodi nudc danh gia tdc dung Hodng HO trong cudn sdch Cay cd Viet Nam, sinh hpc ciing nhu ddc tinh cua loai thyc v4t loai thuc vdt nay phdn bd d Binh Duong, nay. Do v$y, trong nghien ciiu ndy chiing tSii>
Vipt Nam vd Thai Lan. Thdi gian gdn ddy, cd ddnh gid dgc tinh cdp, tac dyng bdo vp gan vJij 14 Tap chi Duac lieu, tap 18, s^ 1/2013
tac d^ng dpc trfin mdt sd ddng tl bao ung thu.
2. B6i tvrpng vd phirong phap nghien cihi 2.1 Ngi^Sn vat lieu thinghiem • •• - - Vdt lifiu nghifin cCoi: Mau nghifin cuu (5 1^) dugc cung cdp bdi; Sd Y tfi Khdnh Hda thang 8 ndm 2012. Thdn cdy xao tam phdn da
&gc Khoa Tdi nguyfin Dugc Hpu, Vifin Dugc lieu xdc djnh tfin khoa hpc la Paramignya trimera (Oliv.) Guillaum, thupc hp Cam (I^taceae), cdn cd tfin ddng nghia la Atalantia trimera Oliv.
Dich chifit va cdc phdn doan tii dugc lieu, hao gom:
+ Cao thifit' methanol (ky hieu la XTP- MeOH) xao tam phdn. ty lfi lg dugc lieu Id 0,075 g cao dugc'diing de nghien cijru tac dung bdo ve gan in vivo vaxac dinh dpc tinh cap.
+ Cao chifit methanol (XTP-MeOH) vd cdc phdn do^n tir cao chilt methanol, bao gim: phdn dogn n-hexan (ky hieu XTP-Hx), XTP ethyl acetate (XTP-EA), phdn doan butanol. (XTP-BuOH), vd -hoat chdt 6-(3,7- dimethyl-2,6-octadienyl)7hydroxyl-2H-1 - ben20pyran-2-one (1) dugc dung de nghifin ci!ru dpc te bdo in vitro. Hogt chdt 1 dugc phdn lap tir dugc lifiu vd Id thdnh phdn hda hoc chinh cua dugc Hfiu.
- Dpng v|tt thi nghifim: ChuOt nhdt trang chiing Swiss albino, trpng lugng 20 - 22 g, sii diing cd chudt due vd cai, la nhung chudt khie mgnh, dgt tieu chudn thi nghifim do Vi^n Vfi sinh Dich te Trung uong cung cdp.
- Cac ddng tl, bao ung thu: Nghien cuu dugc tien hanh.trfin 5 ddng tfi bao img thu, bao_gdm: ung thu gan (Hep-G2), ung thu dgi trang (HTCl 16), ung thu vii (MDA MB231), ung thU'buong trung (OVCAR-8), vd ung thu tii cung (Hela), dugc cuiig cdp bdi Khoa Dugc, Truang Dai hgc Chosun, Hdn Qudc.
- Hda chdt: MTT (3-(4,5-Dimethylthiazol- 2-yl)-2,5-,d^heinyltetrazolium bromide) mdi tmdng DMEM,.huylt thanh thai bd (fetal bovine serum), .trypsin cua hang Invitrogen
(My); Adriaihyin ciia hang Sigma (My), kit dinh' lugng enzym ALT, AST, bilirubin ciia hang Human ^trc) vd cdc hoa chdt khdc...
- Mdy mdc thifit hi: may dgc ELISA cua hang Thermo Labsystems (Biic), kinh hien vi soi ngugc cua hang Canzese (Diic), may Iy tam ngang cua hang Becman Counter (Mf\ mdy dinh lugng sinh hda ban tir ddng cua hang Scouter (Y), bl dn nhiet ShellLab (My)...
2.2. Phuangphdp tiin hdnh
> Phuong phap thft dpc tinh cap:
- Theo "Phuong phdp xac dinh dpc tinh cap cfta thulc" [1]. Chudt nhdt trdng dugc nudi dn dinh 5 ngay trudc khi tifin hdnh thi nghiem. Dung kim cong dau tu cho chupt udng theo miic lifiu tang dan, mfii lan khdng vugt qua 0.4 ml/ lOg thfi trgng chufit. Tim lifiu tdi da md khdng cd chudt ndo cfta Id thi nghiem chet (LDo) vd lieu tdi thieu de 100%
chupt cfta Id thi nghipm chet (LDioo). Thu thfim 3 - 4 lifiu trung gian giiS^ 2 lieu ndi trfin de xdc dinh LD50. Theo doi va quan sat cdc bifiu hifin vfi hdnh vi, hoat dpng, dn udng, bdi tiet cua chupt va sd chudt sfing chfit trong 72 gid. LD50 dugc tinh theo phuong phap Berhen - Kabber.
> Phuang phap nghifin cuoi tdc dyng bdo ve gan:
- Gay viem gan cap cho chupt nhat trang bang paracetamol [8], Dinh lugng hoat dp enzym ALT, AST, hdm lugng bilirubin todn phdn trong huyet thanh de danh gid tdc dyng cua mau thu.
- Tien hdnh thi nghiem:
Chudt thi nghifim dugc chia ngau nhifin thdnh 6 Id: 1 Id chung sinh ly, 1 Id chumg benh ly, 1 Id udng cao chiet methanol lifiu tuang duong 10 g/kg (XTP 1), 1 Id udng cao chifit methanol lifiu tuong duong 20 g/kg (XTP 2), 1 Id udng cao chifit methanol lilu tuong duong 50 g/kg (XTP 3) vd 1 lfi udng silymarin 100 mg/kg (silymarin). Cho chufit cac Id chiing sinh ly vd chiing bfinh 1;^ udng
Tiip chi Duac lieu, tap 18,561/2013- 15
dung dich tween 80 1%, cac 16 thir u6ng Cao chilt hoSc silymarin (pha trong tween 80 1%) tir ngay thil 1 din ngay thii 8 vol thi tich tucmg duong nhau (0,4nil/20 g chupt). Gay viem gan cap bang each cho chugt tat ca cac 16 (trtt 16 ehtag sinh ly) u6ng paracetamol pha trong cacboxy methyl cellulose (CMC) 1% vao ngay thii 7 voi lilu tuong ducmg 400mg/kg thS trong chuot (0,4ml/20g chupt).
Ngdy thii 8, sau khi cho chupt uong (dung dich tween 80 1% h o ^ mau thii hoac silymarin) 1 gio, lay mau tat ca cac chupt cua cac 16 thi nghiem bang each cat c6, ly tam lay huylt thanh de dinh lupng hoat d0 enzym ALT, AST va hdm luong bilirubin toan phan trong huyet thanh.
> Phuang phap danh gia dpc te bao (theo phuong phap cua Mosmann T. cai tiln [7])
- Nu6i cay te bao: Cac d6ng t6 bao dupc luu giii trong nito long, danh thiic va duy tri trong moi truong DMEM (Dulbeeco's Modified Eagle Medium) co b6 sung huylt thanh thai b6 10%, dung dich Ichdng sinh va khang n ^ 1% (penicillin 50,000 units/L va streptomycin 50 mg/L). Tl bdo dupc nu6i ciy cho phat triin toi miic khodng 70%, thay m6i trucmg sach, te bao nay dupc diing lam thi nghidm.
- Mau thi nghiem: Hoa mdu thi nghiem vdo dung dich DMSO 100% sao cho ning dp glc cia cac mau la 10 mg/ml. Tilp theo pha tam nghien cilu thdnh thang ning dp gim 5 ning d6 100, 50, 25, 12.5, va 6,25 ng/ml dli vcri cao chilt phan doan; ning d6 100, 50, 25, 12.5, vd 6.25 nM dli vol chit tinh khilt.
N6ng dp cua mau nghien eta dugc dung theo tieu chuan sang lpc thulc chlng ung thu co nguin glc dupc liSu cua Teicher 1997 [6].
Adriamycin dugc su dung ldm ehtag duong, vcri ning d$ thu lin lupt Id 20; 4; 0.8; 0.16;
vd 0,032 ng/ml. Dung dich DMSO 0,1%
dupc Sll dung ldm ehtag am.
- TiSn hdnh thi nghiem: them vdo cac
gilng cua dia 96 gilng mgt lupng te bdo;
thich hgp v<Si ttog dong t l bdo da dugc hojt h6a 6r tren. Cac t l bao dugc nu6i trong m6i truang 6 37°C va 5% CO2 trong 24 gia. Sau 24 gid nu6i ciy, them 20 pi thulc thu vd thu6c chuin a cac ning dp khac nhau vao mSi gilng. Moi ning d§ dugc ldp lai a i- gilng. li dia nu6i cdy 48 gio (37°C, 5%
CO2). Them 20 pi dung djch MTT vdo moi gi€ng ciia dia nu6i cay vd li 37''C trong 3 gio dl MTT dupc chuyin hoa. Loei bo mfii trudng trong cac gilng ciia dia nuoi cay vd hodn trd fonnazan (sdn phim chuyin hoa MTT) bang 100 pi DMSO. l i e kjf dl forma2an co the tan hoan toan. Cu6i cung, doc mat dp quang a buoc song 540 nm. Mat dp quang se phan anh s6 lupng te bao 56ng sot.
- % tl bao s6ng sau khi da xii ly thuoc so vdi chtog tring dugc tinh theo cong thuc nhu sau:
% ti biho s6ng = IOD540 cua t^ b i o dirgrc xir i^| / IOD540 cua tl b i o khtng dufc xir \f thii6c] x 100%
% ih: chl - 1 0 0 - % te b i o sing
- Cdeh tinh gia tri IC50: Gid tri IC50 dupc tinh bing phdn tich hii quy kh6ng tuyln tinh cua dutog cong dap tag lieu tucmg tag, su dung phan m6m Exell.
3. Ket qua v^ ban lu$n
Theo kinh nghiem dan gian, nguoi dan thutog sic nuoc xao tam phan dl ulng, tuy nhien vol cdeh nay chi co thi chilt dupc cac chat phdn cue. Do v^y, trong nghien eta ndy, ehtag tdi lira chpn cao chilt methanol xao tam phdn voi muc dich co thi chilt dugc phin lto cac chat phdn cue va it phan cue. Cao chiet methanol nay dupc su dyng dl danh gia dpc tinh cdp vd mOt sl tac dung sinh hpc.
3.1. Kit qud thu 36c tinh cua cao chiit methanol xdo tam phdn
Khi cho chu0t ulng din lilu 329g dugc lieu /kg thi trgng chupt, tit cd cac chupt diu khoe manh, ho?t dtog, an ulng, bdi tilt binh thutog, kh6ng c6 chupt nao chit.
16 Tap chi Duac liiu, tip IS, sS 1/290^
Khi cho chudt cdc Id uong vdi cac muc lifiu 411,0; 493,0; 493,0; 657,0 g dugc Hfiu/
kg th^ trpng chupt, t^t cd cdc Id diu chi cd mpt chupt chit trong cd qua tdnh theo ddi.
Cdc chuOt cdn lai dfiu khde manh, hoat ddng, an udng, bdi tifit binh thudng. Nhung chupt chfit dugc md bung dl quan sdt dai thfi cac co quan nfii tang: khdng thay cd bieu hifin bat thudng trfin cdc co quan ndi tang nhu: tim, gan, phfii, d^ day-rupt, thdn...
Sfi H$u vfi thft ddc tinh cdp cfta cao chiet methanol xao tam phan theo dudng udng tren chuOt nhdt trdng dugc trinh bdy d Bdng 1.
BSng 1. S6 li$u thO' d^c tinh cap cua cao chiet x^o tam phan
Kfit qud bdng trfin cho thay: LDo cfta cao chifit methanol xao tam phan nhd hon ho§c bdng 329,0 g dugc lieu/ kg thfi trpng chu0t.
Khi cho cac Id chufit udng lifiu cao han, thdm chi dfin lifiu 657,0 g/ kg thfi trpng chufit Id lifiu t6i da cd thfi cho mdt con chufit nhdt udng dugc mdi chi cd 1/ 10 chufit chet. Nhu vdy khdng xac dinh dugc LD50.
Lifiu nghifin cuu trfin chugt la 329,0 g dugc lieu/ kg thfi trpng quy ddi tuong duong tren ngudi la 27,4 g dugc lieu/ kg thfi trpng hay 1370,0 g dugc lipu/ ngudi n ^ g 50 kg/
ngdy, khong cd bifiu hipn dpc tinh cdp. Tuy nhifin Hfiu ndy lai Id lieu can vdi Hfiu gay ddc.
Vi vay Hfiu tuong ddi an todn (Ds) Id 274,0 g dugc lieu/ ngudi/ ngdy (Ds = l/5LDo) [2].
Tren md hinh ddc tinh cap chi xac dinh dugc sfi chugt sdng chet vd quan sat cac bifiu hifin bfin ngodi cung nhu bfin trong bdng mat thudng. Do do, dfi danh gia dp an todn cfta dugc Hpu ndy cdn cd nhung nghien cftu sdu hon nhu: dpc tinh ban trudng difin, dpc tinh tnrdng difin trfin dfing vat, dp an toan trfin ngudi...
3.2. Ket qud tdc dung bdo vS gan ciia cao chiet methanol XTP
Bfing 2. T&c dyng lie ch^ sy tSng ho^t dp enzym ALT, AST v& h^m lugng bilirubin huylt thanh
cua cdc mlu nghiSn cCru STT
1 2 3 4 5 6
Lilu thir (g dugc li§u/kg the trcing chu^t)
247,0 329,0 411,0 493,0 575,0 657,0
S6 chu9t thii' (con) 10 10 10 10 10 10
si chu9t c h i t (con) 0 0 1 1 1 1
^•'•^~,^^^ ThSng s6
Chiing sinh ly (n = 10) Chiing b^nh IJ (n=16)
XTP 1 (n=12) XTP 2 (n-14) XTP 3 (n=18) Silymarin(n=18)
ALT (U/1) M ± S E 89,4 ±52 445,4 ± 63,5 492,4 ± 69,0 448,3 ± 8,4,2 325,0±51,8«
339,1± 38,7*
% lix che
-10,5 -0,7 27,0 23,9
AST (U/1) M ± S E 133,5 ± 9 , 3 827,3 ± 126,4 611,4 ± 86,8 560,5 ± 108,0 405,2 ± 82,3»
439,9 ± 61,5*
% lie ch^
26,7 32,2 51,0 46,8
Bilirubin (^mol/i) M ± S E 16,9 ± 0 , 5 22,3 ± 0,9 22,0 ± 1,1 22,1 ± 1 , 1 21,8 ± 0,9 2 1 , 7 ± 0.7
% lie c b i
1,4 0,6 2,0 2,4
•: P < 0.05 so vdi 16 chiing b^nh ly
Ket qua Bdng 2 cho thdy: Khi cho chudt udng cao chifit methanol vdi lifiu tuong duong 10 g vd 20 g dugc lieu/kg thfi trpng chupt/ ngdy (XTPl vd XTP 2) trong 8 ngdy lifin tuc, khdng thdy cd s\r thay ddi cd y nghia
thdng kfi cdc enzym ALT, AST va hdm lugng bilirubin huyfit thanh so vdi Id chftng benh ly khdng dung thufic m^c dft ho^t tinh enzym AST gidm ldn lugt Id 26,7% vd 32,2%
(P > 0,05).
Tap chi Duac liiu, tdp 18, so 1/2013 17
6 lilu ulng tuong duong 50g dupc lieu/kg thi trpng ehu6t, XTP dS lie chl 27,0
% su tang hoat dp ALT va 51 % sv tang lioat dp AST huylt thanh chupt dat y nghia thing ke so voi 16 chtog gay benh tren m6 hinh viem gan cip bing paracetamol (P < 0,05), tuy nhien chua lic chl su tang ham lupng bilirubin huylt thanh. D6i vcri da sl cac thit nghiem danh gia tac dung dupc 1^ cua dupc liSu, lilu sit d\mg nghien cto thutog tir 5-20 g dupe lieu/ kg thi trpng chupt. Nhu vay lieu 50g dupe lieu/ kg thi trpng chupt la lilu rit
BSng 3. Ket qud sSng Igc ddc tinh cua cdc
cao. 6 lilu cao nay, xao tam phan ciing the hiSn tao dung ilc chl su ttog enzym gan tr6n m6 hinh gay benh thuc nghiSin, tuy nMeir day khong phai la dli tupng tilm ntog chd dilu tri viSm gan so voi nhilu dirpc lieu khic' da dupc nghien cto.
3.3. Kit qud tdc dung S^c trln mpt so dong ti bad ung thu cua XTP
4 phan doan chilt (100 ng/ml) va 1 hoat chit (100 nM) cOa xao tam phajj bude diu dupc sang lpc doe tinh tren 5 dong te bao ung thu. Kit qua sang lpc dupc thi hien 6 Bang 3.
mau nghien ciiu trgn 5 d6ng tS bdo ung thu
STT
1 2
3
4
5
«
7
K^ hiSu mau
DMSO 0,1%
Adriamycin (20 |ig/ml) XTP-MeOH (100 iig/ml) XTP-Hx (100 ng/ml) XTP-BuOH (100 ng/ml) XTP-EA (100 ng/ml)
XTP-1 (100 nM)
% ij-c ch^ tren cfic d6ng t^ bdo
Hep.G2 0 77,40
87,74
92,72
3,53
7.53
90,79 HTCl 16
0 80,01
97,01
96,57
-5,30
28,84
97,10 MDA MB231 0 85,05
94,43
93,83
-11,3
52,77
OVCAR-8
0 75,15
83,03
87,01
3,95
13,16
94,02 j 90,65
i
Hela
0 95,01
86,91
90,96
-15,97
39,21
90,38
K i t iu|n
Am tinh Duong tinh tr€n 5
dong ti bdo Duong tinh trfil^
d d n g t g b i o ' "
Duong tinh trSn 5 ddng ti bdo
Am tinh
Duong tinh trfin f dong tl bdo Duong tinh trfin 5
d6ng tl bdo
NhOng mau co kha nSng uc chl > 50% su phat triin ciia tl bao ung thu thi dupc lira chpn cho nghien cto tiep theo nhim xac dinh ICso.
Ket qua buoc dau sang lpc cho thiy chi c6 duy nhit phan doan XTP butanol la kh6ng thi hien tac dung dpc tren 5 dong tS bao ung thu kl ti«n. Con Iai cac phan doan XTP Hx, XTP MeOH, va XTP boat chit diu thi hien dpe tinh tren ca 5 dong tl bao. Ph&i doan XTP EA chi
the hien dpc tinh tren mpt dong te bk» ung thu vu (MDA MB231). Theo Michael R.B.[6], nhtog cao chiet th6 dupe sang lpc budc dau it ning dp 100 fig/ml ma co kha nSng diet >50%
tren 3 dong tl bao tmg thu se dupc bfit chpn dl nghign cto sau hon v8 tac dung chlng ung thu.
Dira tren kit qua sang lpc ban diu d tr6n, chung toi tiln htoh xic dinh gia tti ICso cua cac mlu CO kit qua duong tinh.
18 Tgp chi Duac liiu, tdp IS, s61/2013
Bing 4. Gi4 tri ICM tr6n ciia cic mau thu trfin 5 d6ng t^ bSo
K ; hifiu mlu thiy XTP-MeOH XTP-Hx XTP-EA XTP-1 (nM) Adriamycin (nM)
• D6ng tl bdo (Gia tri IC„, ng/ml) Hep-G2
>50 , - 39,61
- -
>50 1,50
, HTCl 16 49,59 76,27
-
55,78 0,50
MDAMB231
>50 62,48
>50 48,74
1,82
OVCAR-8 61,04 51,60
^ -
57,7 1,21
Hela
>50 57,5.
-
18,4 1,48 Kfit qud nghifin curu chi ra rang chi cd
phdn doan XTP-Hx thfi hifin doc tinh tfi bao d mftc do trung binh (IC50 = 39,6,1 p.g/ml) tren dong tfi bao ung thu gan (Hep-G2), cdn lai cac phdn doan khac cho thay tac dung ddc te bao la tuong ddi yfiu. Cdc cHat cd gid tri IC50 nhd hon hoac bdng 5 fig/ml ddi vdi chdt tinh khifit dugc coi Id cd hofit tfnh. Hoat chdt phdn l^p tft xao tam phdn da chi ra cd tac dung ftc che su phat trifin cua te bdo ung thu tft cung d mftc do trung binh (IC50 = 18,4 |j.M tuong duong vdi khoang 5^36 ^g/ml).
Doi vdi nhiing phdn doan cd gia tri ICso>50 ^ig/ml, muc dp ddc tfi bao la tuong d6i yfiu. Tuy nhien, ddy mdi chi la nhihig nghien ciju sdng lpc tac dung dgc trfin mfit s6 ddng tfi bdo ung thu in vitro, de khdng dinh chinh xac ve tdc d\ing nay, can cd nhQng nghifin cuu sdu han trfin dfing vat, ddnh gid ca che tac dung cung nhu ve lam sdng...
Vi$t Nam Id dat nudc nhifit ddi, cd ngudn tai nguyfin thifin nhifin phong phu vd da dang.
Theo tiifing ke t^i Vien Dugc lifiu giai doan 1961-2005, cd khodftg gdn 4000 loai thuc vat da dugc sft dung Id nguyfin lieu lam thudc, trong do cd rdt nhifiu lodi thuc vat da dugc sft dyng trong ho trg difiu tri ung thu, trong dd chua ke dfin cd rdt nhieu bai thudc da dugc luu truyfin trong ddn gian. Mfit sd cdy thudc da cho thdy khd nang gay ddc te bdo ung thu rat meinh, vi d\i nhu cao chifit methanol tft la kho sdm mfim Brucea mollis Wall, ex Kurz uc ch^ su phdt trifin cua tfi bdo ung thu gan (Hep-
G2) vdi gid tri IC50 ^ 14,49 ng/ml [5]. Cdc phdn doan n-hexane tu cao methanol ftc ehfi su phdt trien cfta te bdo ung thu phdi (LU), ung thu gan (Hep-G2), ung thu vu (MCF-7) vdi gia tri IC50 ldn lugt la 3,5 \ig/m\, 1,03 ng/ml vd 5,8 [ig/ml. Cao chifit methanol tft todn cay nhan chdy (Dasymaschalon rostratum var. glaucum (Merr. & Chun) Ban) va than, la cay chudi con chdng (Uvaria grandiflora Roxb. ex Homem) Id 2 cay thudc cua dfing bdo Pako, Quang Tri ciing the hifin dpc tinh rdt manh trfin tngt so ddng tfi bdo ung thu nhu te bdo ung thu bifiu md (KB) vdi gia tri IC50 ldn lugt Id 17,83 ng/ml vd 1,67
|ig/ml; te bao ung thu phdi (LUI) IC50 Idn lupt Id 16,48 ng/ml va 6,86 ng/ml [3]. Chdt oxostephanin dugc phdn lap tft loai binh voi Stephania brachyandra Diels cd hoat tinh ddc manh trfin cac ddng tfi bdo ung thu gan (Hep-G2), ung thu phoi (LU), ung thu mdng tim (RD) vdi.gid tri IC50 rat thdp, ldn lugt Id 0,566; 1,404; vd 0,755 ng/ml [4]. Nhu vdy, neu so sdnh tdc dung ddc tfi bao cfta cdc phan doan vd hoat chdt xdo' tam phdn dd phdn lap dugc vdi cdc chat va phan dofin tu nhung cdy thudc ke trfin thi xao tam phdn khfing phdi la ddi tugng noi bat.
4. Ket luan
- Vfi dgc tinh cdp cao chifit methanol xao tam phan: BChdng xac djnh dugc LD50. Xdc dinh dugc LDo < 329,0 g dugc lieu/ kg the trpng chufit nhdt trang, dftng dudng udng.
- Cao chifit methanol xdo tam phdn khdng
Tap chi Duac lieu, tdp 18, so 1/2013 19
thi hien tae dung to chl sir tang enzym gan bao ung thu gan Hep-G2 (ICso = 39,61 tren mo hinh gay viem gan cip thuc nghi#m ^g/ml) va dong t l bao ung thu c6 tflr cung bing paracetamol 6 chupt nhit tring vcri lilu (ICso = 18,4 pM). Tuy nhien, day m<5ri chi Ii 10 va 20 g dupc lieu/ kg thi trpng chupt. nhtog nghiSn cto bucrc diu de ddnh gid d$c - v l tdc dung dpe tren mpt sl ddng tl bdo tinh ctog nhu mpt sl tac dtog sinh hpc ciia ung thu; Cao ehilt methanol, phan doan n- xao tam phin. Cin co nhtog nghien cihi hexan va hoat chit tir xao tam phto thi hipn chuyto sdu hon, co sf phii hpp ciia nhilu co d§c tinh tren ca 5 dong tl bao ung thu, trong quan nghien cto trong va ngoii nudc de d6 phin doan n-hexan va hoat chit thi hipn khing dinh chinh xac tic dung cia cay thulc dpc tinh o rnilc dp trung binh, lin lupt trto tl niy.
Tii li$u tham khdo
I. Do Trung Dim (1996), Phuong phSp xdc dinh d^c tinh d p cua thufic. Nxb Y hoc - Ha NOi. 2. Vi^n Dmrc li^u (2006), Phirong phap nghien cijru tic dung dugc IJ ciia thu6c tir thao duoc, N X B Khoa hpc vk Ky thuyit, tr 352- 3 6 3 . 3 . Nguyen Thj Hoai, Tnnh Thj Diep, DS Thj Thio, Nguyen Khinh Thiiy Unh, Nguyin Bich Hiln, Hoing Thi Di«u Huong (2012), Sing loc hogt tinh di§t l l bio ung thu mpt s6 cSy thu6c ciia d6ng b i o pako, vin ki^u * Quing Trj. Tsp chl Dugc li?u, 2:95-100. 4. Nguyin Qu6c Huy, Ph?m Thanh KJ, LB Mai Huong (2009), NghUn ciru tic dyng gay doc tl bio ciia 10 hop chat phSn l|p ttr m$t sfi Ioii binh voi {Step/tania Lour.) fir Vi§t Nam, T*p chl Dugc lisu 4:218-220. 5. Mai Hilng Thanh Timg, Tran Thu Huimg, Trin Thi Bich, Nguyin Qufic Blnh, B4 Thi Cham, Nguyen Manh Cuimg (2011), Tic dyng giy d0c tl b i o ung thu cua l i khfi Sim mim. Tsp chi Dugc li^u, 6:356-360. 6. Beverly A. Teicher (1997), Anticancer drug development guide: Preclinical screening, clinical trials, and approval. Humana Press, Totowa, New Jersey. 7. Mosmann T. (1983), Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods:
65, 55-63.8. Hsu CC, Lin KY, Wang ZH, Lin WL, Yin MC (2008), Preventive effect of Oanoderma amboinense on acetaminophen-induced acute liver injury, Phytomedicine, 15(11), 946-950.
Ttfp chi Dtrcrc lieu, tap IS, so 1/2013 (Trang 20 - 25)
NGHIEN C U t J T A O M O S E O T R A M T R A VA N H A N B I E T H A M L i r O N G T E R P I N E N - 4 - O L T R O N G M A U N l J O I C A Y
'Trin Van Minh, 'BS Ttin Vinh, 'Nguyin Thien Quang Dgi hoc Nguyin Tdt Thdnh, 'Trudng Dgi hgc Quic ti'Dgi hqc Qudc gia
Thdnh phi H6 Chi Minh (Nhjn bai ngiy 20 thdng 11 n5m 2012)
Summary
Study on Callus Induction of Tea Tree {IHelaletlca altemifolia) and Identiflcation of Terpinen-4-ol Content in Cultures
The leaves explants of in vitro cultured Melaleuca alterni/olia plantlets were used as initial materials for cultures. The leaves were cut horizontally into two pieces and cultured on MS (1962) medium containing 2.4-D (4 mg/l), BA (0.5 mg/l), sucrose (20 g/l), vitamine Bl (5 mg/l) and glycine (10 mg/l) ) in dark at 26+2°C Sir callus induction. Alter 40 days of culture, the rapidly growing white and soft calli were cultured on MS medium supplemented with 2.4-D (1 mg/l), sucrose (20 g/l), coconut water (15% v/v), glycine (10 mg/l) and vitamine Bl
20 Tap chi Duac lieu, tdp IS, sg 1/20^
