• Nghien CCFU - Ky thuat
Khao sat tac dong ha glucose huyet
r
va doc tinh cap cua la dua thom (Pandanus amaryllifolius Roxb.)
Le Thi Ngoc Anh, Huynh Ngpc Trinh*
Bo mon Duoc Ly, Khoa Duac, DH Y Duoc. Thdnh pho Ho Chi Minh
*E-mail: trinhbl8I@yahoo.com
Summary
The leaves of Pandanus amaryllifolius (P.A.) of time-honored use in folk-medicine for diabetes mellitus were investigated to elucidate the hypoglycemic activity by the model of alloxan-induced type 2 diabetic mouse. The total plant extracts (of P.A.) at doses of 25 g and 10 g/kg b.w remarkably lowered the blood glucose level of treated mice as compared to untreated ones. The hypoglycemic effect was not significantly different from that of gllbenclamide 5 mg/kg. fvforeover, the P.A. extracts exhibited no acute toxicity at the maximal oral dose given to mice (100 mg/kg). These clarified the potential use and supported the safety of P.A. extracts in the treatment of diabetes mellitus.
Keywords; Pandanus amaryllifolius, diabetes mellitus, alloxan, gllbenclamide.
Dat van de
Trong nhdng nam gan day, ngydi dan ed xu hydng s d dung cac cay thuoc theo kmh nghipm ddn gian d l kiem soat glucose h u y l t Cae eay thuoe ndy da gdp phan hdu hipu trong dieu tri bpnh ddi thdo dydng (OTO) dong thdi giam d y g c chl phi cung nhy cdc tac dung phy eua eae thudc tan d y g c Trong s l do, la dda tham (cay cam n i p ) Pandanus amaryllifolius Roxb., hp Dda gai Pandanaceae'^'eung thydng d y g c s d dung d l ha glucose huylt. Tuy nhidn, cac nghien cdu trong nyde ve tde dung d y g c ly cua la dda tham v l n cdn rat han e h l . Mdt s l nghien edu tren t h l gidi cho thay edy Pandanus amaryllifolius Roxb. ed tde dyng ha glucose huylt vd ed the nhdm hgp chat steroid mang lai tac dyng nay '^. Ngodi ra.
Id Pandanus amaryllifolius Roxb. cdn the hidn tde ddng khang khuan in vitro va gay ehit t l bao ung thy vii ddng MDA-MB-231 in vitro P*'. Do do.
nham ehdng minh tac dung hg glucose h u y l t cua Id dda tham, chiing tdi da thye hien d l tai "Khao sat tae ddng hg glucose huyet va dde tinh edp eua id dda tham {Pandanus amaryllifolius Roxb./'
Vat lieu va phu'ang phap nghien CU'U
50
Vdt lieu
Dong vdt thw nghiem
Chupt nhdt trdng chiing Swiss albino, giong dye, can nang khodng 25 - 30 g, d y g e mua td Vidn Vac xin va Sinh pham y t l Nha Trang va d y g e nudi bang thde an vidn do Vi^n cung d p . Chupt d y g e nudi on djnh d nhipt dp phdng 3-4 ngdy t r y d c khi bat dau moi thd nghipm.
Hoa chat
Alloxan (Sigma-Aldrieh Chemie Gmbh);
glibenclamid 5 mg (Cdng ty Lien doanh TNHH STADA); thuoc t h d glucose (iSE, Italy); dung dieh NaCI d y g c dung; Dung mdi c h i l t x u l t : con 50 % (dung mdi ky thudt Trung Quoe).
Cao todn phan la dwa thom
La dda thom d y g c thu hai tgi Gd Vap - Thdnh pho Ho Chi Minh. D y g e lipu d y g c tdch la, n>a sach, phai am can va xay nhd d l ehiet Dyge lieu nghien edu d y g e ehilt xuat theo phyang phdp ngam kiet vdi dung mdi eon 50 % theo ti 1^
1:10. Dich ehilt thu d y g e d y g e cdtren b i p edeh thiiy 70 °C den eao dge. S l y ehan khdng cao ddc khoang 7 ngay d y g e eao todn phan. Bao quan eao trong binh hut am de trdnh hut I m . T A P C H i DlTQC H Q C - 7/2015 ( S 6 471 NAM 55)
Nghien ciru - Ky thuat
Khao sat tdc dung ha glucose huylt Chupt d y g c eho nhjn ddi qua ddm t r y d e khi tidm alloxan. Chudt d y g e gay tang glucose h u y l t bang each tiem tlnh mgeh dung djeh alloxan l i l u 50 mg/kg (pha trong dung djch NaCI 0,9 %). Q thdi diem 48 g i d sau tidm, ehudt d y g c lay mau d l danh gia tinh trang DTO. Nhdng chudt cd nong dp d y d n g huydt cao han 200 mg/dL d y g e lya chpn vao thi nghidm khao sat tde dyng ha glucose h u y l t ciia cae cao d y a c lieu.
Chudt bj dai thdo d y d n g d y g e didu tri ngau nhidn bdng each cho udng thuoc ddi ehdng glibenelamid (lidu 5 mg/kg, 1 Idn/ngay) hoge eao la dda tham (lidu t y a n g d y a n g 10 g va 25 g bot d y g c lidu khd/kg, 1 Idn/ngay). Nhom ehudt chdng bpnh d y g c cho uong n y d e c i t (10 ml/kg, 1 lan/ngay) trong s u i t qud trinh thd nghidm.
Chupt d y g e dieu tri trong vong 15 ngdy Theo ddi glucose h u y l t eua chudt moi 5 ngay. Mau dygc lay vdo b u l l sang t r y d c khi cho chudt an.
Thdm dd dpc tinh cap
Cao toan phan la dda thom d y g c nhu hda trong Tween 80 (4 %), pha dung djch ddm dae toi da cd t h l bam d y g e qua kim eho chudt uong (tyang dng 20 g bdt d y a c lieu khd/ml). Chudt dygc cho nhjn ddi 12 gid nhyng cho uong n y d e
ddy dil t r y d c khi t i l n hanh thf nghipm. Dung kim ddu tu cho chudt uong mdt lidu duy nhdt thudc t h y nghiem. Gdm cac nhdm chudt nhy sau:
Nhom chung: udng n y d e cdt 10 ml/kg.
Nhom dung moi: udng Tween 80 (4 %) 10 ml/kg.
Nhom thw udng eao toan phdn la dda tham vdi lieu t y a n g dng 200 g bdt d y g c lieu khd/kg.
Theo ddi s y thay doi hanh vi, t h l trang chudt va tl lp ehupt e h i t trong vdng 72 gid. Tinh LD^^
theo phyang phap Karber va Behrens. Mo quan sat dai the nhdng chudt c h i t trong liic t h d nghipm vd sau khi k i t thuc dgt t h d nghipm
Phu'O'ng phdp ddnh gid k i t qua Cac sd lieu thu d y g c d y g c trinh bay d y d i dang Trung binh ± SEM. Dimg phdn mdm Minitab 16 de SO sanh s y khac bipt gida eae ngay d i l u tn so vdi ngay ddu trong cimg mpt Id va gida cac Id khae nhau S y khac bidt cd y nghTa khi p < 0,05.
K§t qua nghien CLFU
Tac ddng ha glucose huylt cua cao todn phdn Id dda tham
Glucose huyet trung binh eiia cdc nhdm ehudt t h d nghidm trong qua trinh d i l u tn d y g e trinh bay trong bang 1.
Bang
Sinh ly Chiing b^nh Glibenclamid 5 mg/kg Cao ia dtra tham 25 g/kg Cao la dija tliom 10 g/kg
| 1 : n
8 10 8 7 8
N&ng do glucose
NgayO 112,9 ±4.3 397,0 ±40,4 326,1 ±36,2 282,3 ±37,6 423,0 ±29,3
huyet trung binh (mg/dl) cua cac nhom chuot Nong do glucose huyet (mg/dl) Ngay 5
122,9 ±6,9 419,6 ± 31,2 300,8 ±36,6 217,7 ± 34,5 370,8 ±49,8
Ngay 10 124,2 ±5,7 351,0±29,7 211,9±22,20 159,2 ±12,40 2394 ± 32,70
Ngay 15 120,1 ± 5,9 389,0 ±26,1 131,8 ±7,70 143,2*12,40 2034 ±23,20 (*) p<Q,Q5 khdc biet cd y nghfa so vai ngay dau dieu tri trong cung mdt nhom
K i t qua thye nghidm eho thay d lieu 50 mg/kg tidm tinh mgeh dudi, alloxan gay tinh trang bdnh tyang doi I n dinh trong vdng 15 ngdy didu trj.
Sau khi dieu tn vdi glibenclamid hay vdi cao toan p h l n Id dda, glucose huyet eua ede ehudt thyc nghidm da giam d i n so vdi thdi d i l m ban dau.
Sau 5 ngay d i l u tn nong dp glucose huyet ciia
cac nhdm chudt cd giam nhp nhyng ehya khac bidt cd ^ nghTa thong kd so vdi ngay dau dieu tn (hinh 1). Glibenclamid 5 mg/kg iam giam 8 % trong khi eao la dda thom lieu 25 g va 10 g/kg lam giam lan l y g t 23 % vd 12 % mde glucose h u y l t so vdi thai diem ban ddu.
TAP CHi DlTQC HQC - 7/2015 (SO 471 NAM 55)
Nghien ciru - Ky thuat
200 - TOO -
—LD 25 g/kg —*—LDlOg/kg —•*—Gii Smg/kg Hinh 1 : Glucose huydt cua nhom chuot thw nghiem
C) p < 0,05 so vifi ngay ddu dieu tri cOa cung nhom chuot; LD: cao la dira thom; Gii: glibenclamid Tuy nhidn, vao ngdy 10 va 15, glibenclamid
cung nhy cao Id dda tham da lam giam dang k l glucose h u y l t ciia chudt t h d nghidm va khac biet cd y nghTa so vdi ban dau. Vao cuoi thd nghidm, nong dp d y d n g h u y l t eua eae nhdm giam l l n l y g t la 60 %, 49 % vd 52 %.
Ngoai ra, khi so sanh vd hieu qua ha glucose huylt, k i t qua phan tich thong kd cho t h i y khdng cd s y khae bipt cd y nghTa gida 2 lieu nay eua cao la dda tham va nhom glibenelamid 5 mg/kg (p > 0,05).
Doc tinh cap cua cao Id dda tha'm tren chupt binh thu'dng
Sau khi u l n g nyde cdl hay dung dich Tween 80 (4 %), cac chudt nhdm chdng cung nhy nhdm dung mdi v l n hoat ddng va an uong binh t h y d n g . Trong khi dd, chudt d nhdm t h d giam hoat ddng, di lgi chgm ehap, mpt vai ehugt ndm eo lai, cd ve met mdi, khdng an udng trong 10 - 1 5 phut dau.
J-N^
Sau khoang 5 - 6 gid, chupt ed b i l u hipn tidu ehdy, phan den va keo dai trong khoang 24 gid dau tidn. Chugt met mdi, di lai eham chgp. Sau 24 gid, ehugt b i t ddu lmh hoat trd Igi, an udng kha han, hidn t y g n g tidu chay eham dirt vd trd v l trang thai binh t h y d n g , khdng ed b i l u hlpn gi khac t h y d n g . Sau 72 gid, chudt hogt ddng binh t h y d n g va khdng ed chugt e h i t .
Khi mo chupt de quan sat dgi t h l sau 72 gid, ehung toi nhgn t h i y npi tgng eua nhdm chdng va nhdm dung mdi khdng cd bieu hipn khae thydng:
tim binh t h y d n g , phoi trdng hdng, toan bp la gan ed mau do t y a i , djeh mat vang t y a i , tui mat dly, rugt binh t h y d n g . Nhom ehugt thy eo tong t h l npi tgng vdn binh t h y d n g . Tuy nhidn, ehudt c6 mpt so b i l u hipn nhy; rupt hai to han so vdi chupt binh t h y d n g , dg day phinh to, dich dg ddy cd mau hai xanh.
- " • s ^ .
Chudt chung Chugt udng Tween 80 (4%)
Hinh 2; Dai the ngi tang chugt thw nghiem doc tinh cdp Ban luan
Chuot uong cao Id dCra tham 200 g/kg
Nhy vgy, eao ehidt toan phan eon 50 % t d id dda tham khdng gay d i u hidu dgc tfnh edp d lieu 200 g bpt dyge lieu khd/kg va ehya xae djnh d y g c lieu LDjp eua eao todn p h l n la dda tham khdo sat.
Oieu nay eho t h i y tinh an toan cua cao la dda tham va kha nang dng dyng cao nay trong d i l u trj BTO.
Vdi mill tham dgc tryng, t d lau trong dan gian la dda tham {Pandanus amaryllifolius Roxb.J da d y g e s d dyng d l tao mtii tham, tgo mdu va hyang lipu. Gdn day tren t h l gidi cd mot so
52 T ^ CHI DlTQC HQC - 7/2015 (SO 471 NAM 55)
Nghien CLPU - Ky thuat
cdng trinh nghien edu khac nhau vd cdng dung eija la cay Pandanus amaryllifolius Roxb.; trong do, thanh phdn tham 2-aeety!-1-pyrrrolin d y g e quan tam nghidn cdu dng dyng trong nganh hda thyc pham. Mdt nghien cdu khac eiia Fatihanim Mohd Nor va CS nam 2008 da cho thdy tidm nang s d dung djch ehiet Id cay Pandanus amaryllifolius Roxb. nhy mgt e h i t chdng oxy hda t y nhien i^. Ting-ting Jong eung edng s y da phan tfeh thanh p h l n hda hpc eua cac bd phgn g l e ciia Pandanus odoratisslmus ( d y g e cho la gidng vdi Pandanus amaryllifolius) va chidt tach phdn lap d y g c hai hgp e h i t phenolic, b i n loai hgp c h i t lignan va mdt benzofuran mdi. Trong so dd, pinoresinol va 3,4 -bis (4-hydroxy-3- methoxybenzyl) tetrahydrofuran the hidn tac dpng chong oxy hda manh me^''] Ngoai ra, la cay Pandanus amaryllifolius Roxb. cdn cd tac dung hg glucose huylt, khang khuan, khang virus, trj thap khdp...'^'^'
K i t qua dieu tn trdn nhdm ehupt dai thdo dydng cho thay kha nang lam ha glucose h u y l t ciia cao toan phan Id dda tham. Tae dpng nay t h l hipn rd d ed 2 lieu 10 g va 25 g bpt d y g c lieu khd/kg va khdng ed s y khde biet cd ^ nghTa thong kd gida hai mdc lieu nay. K i t qua didu tri nay phii hgp vdi nghien c d u eua Sreenivasan Sasidharan vd cdng s y nam 2011 khi nghien cdu tac dyng hg glucose huyet cua djch ehilt ethanol la eay Pandanus amaryllifolius Roxb.'^l Tae gia cung ehi ra rang, djeh c h i l t ethanol cua la cdy Pandanus amaryllifolius Roxb. cd tae dpng bao ve t l bao trong cac t i l u dao Langerhans cua t u y i n tyy nhd cac thanh phan hda thyc vat trong dich c h i l t ethanol cua la cay Pandanus amaryllifolius Roxb.
giup can bang lygng d y d n g trong mau va gay nen s y tai sinh te bdo npi md t u y i n tyy.
K e t l u a n
Cao toan phan eon 50 % t d Id dda tham lam hg rd ret mde glucose h u y l t eua ede ehupt d y g c gay ddi thao d y d n g thye nghidm, Cao ehilt ndy khdng gdy b i l u hidn dde tinh cap trdn chudt binh thydng u l n g l i l u t l i da cd the bam qua kim cho chudt u l n g (200 g/kg). Nhdng k i t qua tren dem
lai hy vpng v l kha nang tgn dung mdt ngudn nguyen lieu dd tim trong ddi sdng d l phat trien mpt san pham s d dung trong d i l u trj hd tra bdnh dai thao d y d n g .
T a i l i e u t h a m k h a o 1. V6 Van Chi (2002), Td dien cay thuoc Viet Nam, Nxb Y hpc, tr 804.
2. Sukandar D., Sumarlin L. 0,, Zahroh H., Amelia E. K. (2011), "Xenobiotica. Structure elucidation of antidiabetic compound of ethyl acetate extract of pandan wangi {Pandanus amaryllifolius Roxb.) leaves". Proceedings of the 2"'' International Seminar on Chemistry, pp, 240-243.
3. Dumaoal O. S. R., Alaras LB., Dahilan K G., DepaduaA A., PulmonesC. J. (2010), 7n v//ro activity of pandan {Pandanus amaryllifolius) leaves crude extract against selected bacterial isolates.", JPAIR:
Multidisciplinary Journal, 4, pp. 240-243.
4. Chong H. Z., Yeap S. K., Rah mat A., Akim A.
M., Ahtheen N. B., Othman F, et al. (2012), "In vitro evaluation of Pandanus amaryllifolius ethanol extract for induction of cell death on non-hormone dependent human breast adenocarcinoma MDA-MB-231 cell via apoptosis", BMC Complementary and Alternative hAedicine, 12:134
5. F M Mohamed, S. Idris, N.A. Ismail, R. (2008),
"Antioxidative properties of Pandanus amaryllifolius leaf extracts in accelerated oxidation and deep frying studies", Food Chemistry, ^^0, pp 319-327.
6. Ting Jong et al (1998), "Antioxidative activitives of constituents isolated from Pandanus odoratisslmus", Phytochemistry, 49 (7), pp. 2145-2148.
7 Ooi L, S. M , Sun S. S. M„ Ooi V. E. C. (2004),
"Purification and characterization of a new antiviral pro-tern from the leaves of Pandanus amaryllifolius (Pandanaceae)", The International Journal of Biochemistry & Cell Biology, 36, pp 1440-1446.
8. Taylor & Francis (2010), "Chemical constituents and post-harvest prospects of Pandanus amaryllifolius leaves. A review", Food Reviews International, 26 pp. 230 - 245.
9. Sreenivasan Sasidharan, et al (2011),
"Antihyperglycaemic effects of ethanol extracts of Carica papaya and Pandanus amaryllifolius leaf in streptozotocin-induced diabetic mice". Natural Product Research, 25, (20), pp 1982-1987.
{Ngdy nhdn bai: 03/12/2014 - Ngdy duyet dang: 02/07/2015)
T^P CHi DlTQC HQC - 7/2015 (SO 471 NAM 55)
