B Y T TRNG I HC DC H NI INH TH THU THY NH GI TC DNG H GLUCOSE MU CA DCH CHIT L CC LOI TRONG CHI GYMNEMA R.BR. VIT NAM KHA LUN TT NGHIP DC S H NI 2014 B Y T TRNG I HC DC H NI INH TH THU THY NH GI TC DNG H GLUCOSE MU CA DCH CHIT L CC LOI TRONG CHI GYMNEMA R.BR. VIT NAM KHA LUN TT NGHIP DC S Ngi hng dn: 1. TS. Phng Thanh Hng 2. ThS. Phm H Thanh Tng Ni thc hin: B mn ha sinh Trng i hc Dc H Ni H NI 2014 LI CM N Vi lng knh trng v bit n su sc, trc tin em xin by t li cm n chn thnh ti ngi thy hng dn ca mnh TS. Phng Thanh Hng - B mn Ha sinh - Trng i hc Dc H Ni. C cho em rt nhiu kinthcqubutrongphngphphctp,nghincucngnhtrong cuc sng. C lun hng dn tn tnh v to mi iu kin thun li cho em trong sut qu trnh hc tp v nghin cu. EmxingilicmnchnthnhtiThs.PhmHThanhTng- B mn thc vt - Trng i hc Dc H Ni to iu kin cho em c c mu nghin cu v gip em trong qu trnh thc hin kha lun. Em xin cm n cc thy, c gio, cc anh ch k thut vin ca b mn Ha sinh, b mn Dc l to iu kin gip em hon thnh kha lun ny. ng thi em xin cm n gia nh v bn b quan tm, ng vin v gip em trong qu trnh hc tp v thc hin kha lun ny. H Ni, ngy 10 thng 5 nm 2014 Sinh vin inh Th Thu Thy MC LC DANH MC T VIT TT DANH MC BNG DANH MC HNH T VN .............................................................................................. iii CHNG 1: TNG QUAN ............................ Error! Bookmark not defined. 1.1.Tng quan v bnh i tho ng ................................................ 3 1.1.1. Khi nim .......................................................................................... 3 1.1.2. c im dch t ................................................................................ 3 1.1.3. Phn loi ............................................................................................ 5 1.1.4. Tiu chun chn on ........................................................................ 6 1.1.5. Cc xt nghim ha sinh chn on bnh T .................................. 6 1.1.6. Cc bin chng ca bnh T........................................................... 7 1.1.7. Cc thuc iu tr T ...................................................................... 8 1.2.Tng quan v chi Gymnema R.Br................................................. 11 1.2.1. V tr phn loi ................................................................................. 11 1.2.2. c im thc vt v phn b .......................................................... 11 1.2.3. Thnh phn ha hc ......................................................................... 14 1.2.4. Tc dng iu tr T ..................................................................... 15 CHNG 2: NGUYN LIU, I TNG, PHNG TIN V PHNG PHP NGHIN CU ....... Error! Bookmark not defined. 2.1.Nguyn liu ..................................................................................... 18 2.2.i tng nghin cu ..................................................................... 18 2.3.Phng tin nghin cu ................................................................. 19 2.3.1. Ha cht ........................................................................................... 19 2.3.2. Thit b dng chit v pha ch dch chit ..................................... 19 2.3.3. Thit b dng trong qu trnh th tc dng sinh hc ......................... 19 2.4.Phng php nghin cu ............................................................... 19 2.4.1. Phng php chit xut dc liu .................................................... 19 2.4.2. Phng php nh lng glucose huyt ............................................ 20 2.4.3. Phng php gy tng glucose mu bng streptozocin ..................... 20 2.4.4. Phng php nghin cu tc dng h glucose mu trn ng vt th nghim ........................................................................................ 21 2.4.5. X l s liu ..................................................................................... 21 CHNG 3: THC NGHIM, KT QU V BN LUNError! Bookmark not defined.3.1.Kt qu ........................................................................................... 22 3.1.1. Xc nh liu dng thch hp c tc dng h glucose mu trn m hnh chut b gy tng glucose mu bi STZ liu 150 mg/kg ........... 22 3.1.2. Tc dng h glucose mu ca Gymnema sylvestre (Retz.) R.Br. ex Schult ............................................................................................... 25 3.1.3. Tc dng h glucose mu ca Gymnema latifolium Wall. ex Wight . 27 3.1.4. Tc dng h glucose mu ca Gymnema inodorum (Lour.) Decne ... 29 3.1.5. Tc dng h glucose mu ca Gymnema yunnanense Tsiang ........... 31 3.1.6. So snh mc h glucose mu gia cc loi Gymnema R.Br............. 33 3.2.Bn lun .......................................................................................... 34 3.2.1. Tc dng h glucose mu ca Gymnema sylvestre (Retz.) R.Br. ex Schult ............................................................................................... 34 3.2.2. Tc dng h glucose mu ca Gymnema latifolium Wall. ex Wight . 35 3.2.3.Tc dng h glucose mu ca Gymnema inodorum (Lour.) Decne...36 3.2.4. Tc dng h glucose mu ca Gymnema yunnanense Tsiang ........... 36 3.2.5. So snh tc dng h glucose mu gia cc loi Gymnema R.Br. ...... 37 KT LUN................................................................................................ 38 KIN NGH ................................................................................................ 38 TI LIU THAM KHO DANH MC T VIT TT ADAAmerican Diabetes Association Hip hi i tho ng Hoa K BMIBody Mass Index Ch khi c th Ti tho ng GLUT2Glucose transporter Yu t vn chuyn glucose GODGlucose oxidase HDLHigh density lipoprotein Lipoprotein t trng cao IDFInternational Diabetes Federation Lin on i tho ng th gii LDLLow density lipoprotein Lipoprotein t trng thp MODYMaturity Onset Diabetes of the Young i tho ng khi pht lc tr tui STZStreptozotocine WHOWorld Health Orgnization T chc Y t th gii

DANH MC BNG Bng 2.1Cc mu s dng trong nghin cu Bng 3.1S ph thuc liu dng G.sylvestre (Retz.) R.Br. ex Schul v tc dng h glucose mu trn m hnh chut tng glucose mu thc nghim bi STZ Bng 3.2S thay i glucose mu ca cc l chut ung G.sylvestre (Retz.) R.Br. ex Schul Bng 3.3S thay i glucose mu ca cc l chut ung G.latifolium Wall. ex Wight Bng 3.4S thay i glucose mu ca cc l chut ung G.inodorum (Lour.) Decne Bng 3.5S thay i glucose mu ca cc l chut ung G.yunnanense Tsiang DANH MC HNH Hnh 3.1nh hng ca liu dng G.sylvestre (Retz.) R.Br. ex Schul n mc h glucose mu trn chut tng glucose mu thc nghim bi STZ Hnh 3.2T l % h glucose mu ca cc l chut ung G.sylvestre (Retz.) R.Br. ex Schul Hnh 3.3T l % h glucose mu ca cc l chut ung G.latifolium Wall. ex Wight Hnh 3.4T l % h glucose mu ca cc l chut ung G.inodorum (Lour.) Decne Hnh 3.5T l % h glucose mu ca cc l chut ung G.yunnanense Tsiang Hnh 3.6T l % h glucose mu ca cc loi Gymnema R.Br. Hnh P.1 c im hnh thiloi Gymnema sylvestre (Retz.) R. Br. ex Schult Hnh P.2c im hnh thi Gymnema inodorum (Loureiro) Hnh P.3c im hnh thi Gymnema latifolium Wallich ex Wight Hnh P.4 c im hnh thi loi Gymnema yunnanense Tsiang 1 T VN ithong(T)lmtbnhmntnh,kmtheonhiubin chng nguy him trn mt, thn, tim mch, thn kinh, S gia tng t bin v t l ngi mc bnh T ang l mt gnh nng cho ngnh y t ni ring vchotonxhinichung.Cngvibnhtimmchvungth,Tl mttrongbabnhphttrinnhanhnhthinnay.Tchcytthgii (WHO)cnhngcnhbovnguycgiatngTtrntonthgii [30]. V vy, vic iu tr T l mi quan tm ca nhiu nh nghin cu. Vic nghin cu, khai thc v pht trin thuc c ngun gc thin nhin lmt xu hng ngycng pht trin trn ton th gii. c bit, Vit Nam c ngun ti nguyn dc liu phong ph, c nn y hc c truyn lu i. T hngnghincu,crtnhiuloidcliucnghincuv chngminhtcdnghglucosemunh:dythacanh,hongtinh,mp ng, d, [12,20]. ChiGymnemaR.Br.hTrco(Apocynaceae)ckhong25loi, phn b vng Ty Chu Phi, Australia, chu . Trong chi Gymnema R.Br. loiGymnemasylvestre(Retz.)R.Br. ex Schul c nghincu nhiu v tcdnghglucosemuvcs dngrngriiutrTnhiu nc trn th gii. Vit Nam, pht hin s c mt ca mt s loi thuc chiny [1,7].Ngoi Gymnema sylvestre(Retz.) R.Br.exSchul,cc loi cn li hu nh cha c nghin cu v tc dng iu tr T trn th gii. Liu cc loi ny c tc dng h glucose mu hay khng? Mt khc, bn thn loi Gymnema sylvestre (Retz.) R.Br. ex Schul mc d c a vo nhiu ch phmthucvthcphmchcnngiutrTnhngtcdngtra khngnnh,ngnht,cthldongunnguynliuchacchun ha. c th nh gi c tc dng h glucose mu ca cc loi Gymnema v gpphncho cng tc nghincuphttrin,chunha ngundcliu 2 iu tr T trong nc, ti nh gi tc dng h glucose mu ca dch chitlccloitrongchiGymnemaR.Br.VitNamcthchinvi cc mc tiu: 1.nhgitcdnghglucosemucaccmuthucbnloi Gymnemasylvestre(Retz.)R.Br.exSchul,GymnemalatifoliumWall.ex Wight, Gymnema yunnanense Tsiang, Gymnema inodorum (Lour.) Decne thu hi cc vng khc nhau ca Vit Nam. 2.La chn c mt s mu c tc dng h glucose mu u th nht trong chi Gymnema R.Br. Vit Nam c th bo tn, pht trin, khai thc lm nguyn liu. 3 CHNG 1. TNG QUAN 1.1.Tng quan v bnh i tho ng 1.1.1. Khi nim Theo nh ngha ca WHO, i tho ng l mt nhm bnh chuyn hacctrngbitngglucosemumntnhdohuqucasthiu ht hocgim hot ngca insulin hockt hpchai.Tng glucosemu mntnhtrongTlmtnthng,rilonvsuygimchcnngca nhiu c quan khc nhau, c bit l tn thng mt, thn, thn kinh v tim mch [2]. 1.1.2. c im dch t 1.1.2.1. Tnh hnh mc bnh trn th giiTheodoncaWHO,Tstr thnh idchiviccnc angphttrintrongthk 21ny.Hinnay,chn70%sngimc bnh sng cc nc thu nhp thp v trung bnh [29]. -Nm2010,trnthgiic285triucamcT,tngngvi 6,4%dnstrngthnh.Dkinstngln438triucavonm2030, tng ng vi 7,8% dn s trng thnh [29]. -Tlmcbnhtrungbnhtrnthgiil6,4%,caonhtl10,2% khu vc Ty Thi Bnh Dng v thp nht l 3,8% khu vc Chu Phi. Tuy nhin, theo c tnh khu vc Chu Phi s c tc gia tng cc ca mc bnh cao nht [29]. -Hinnay,Tchyuxyranhmtuit40-59.Dkinn nm2030,Tstptrungchyuvonhmtui60-79vi196triu ngi [29].-T l mt trong nhng nguyn nhn chnh gy t vong sm trn ton th gii. Trong cc bnh khng ly th 60% ca t vong l do mc T [29]. 4 Mt vn khc l chi ph cho iu tr bnh T [29]: -TheothngkcaIDF,tngchiphchoTtrntonthgiinm 2010 c tnh khong 418 t USD v s ln ti trn 561 t vo nm 2030. -Bn cnh chi ph y t trc tip trong iu tr bnh T cn tnh n chi phchoiutrccbinchngcabnh.TheodbocaWHO,trong khong10nm,t2005n2015,chiphchobnhtimmchdoTti TrungQucl 557,7tUSD,tiNga l 303,2tUSD,tin l336,6t USD, ti Brazil l 2,5 t [29]. 1.1.2.2. Tnh hnh mc bnh Vit Nam Nhngnmgny,viciutradchtbnhTVitNam c t chc mt cch tng i c h thng. Nm 2001, ln u tin iu tra dch t bnh T ca Vit Nam c tin hnh theo cc tiu chun quc t 4 thnh ph ln l H Ni, Hi Phng, Nng v thnh ph H Ch Minh; i tng nghin cu t 30 n 64 tui. Theo kt qu, t l mc bnh T l 4,0%; ri lon dung np glucose l 5,1% c bit l c ti 64,9% s ngi b T khng c pht hin kp thi v khng c hng dn iu tr [2].Nm2002,disch ocabnhvinnitittrungng,mts aphngtinhnhnghincutnhhnhmcbnhTvccyut nguy c mc bnh a phng mnh. Cc nghin cu ny u cho thy bnh T tng nhanh khng ch cc khu cng nghip m cn c vng min ni v trung du. Vd thnh ph Yn Bi,tlmcbnh trn 3,0%, trong c gn 70% ngi mc bnh T cha c pht hin kp thi [2]. Nm2010,theothngkcaIDF,tiVitNamc1.646.600ngi mc bnh T, chim 3,5% dn s c nc. Con s ny d kin s tng ln 4,4% nm 2030. Nm 2010 s ngi t vong do T l 32.505, tng chi ph cho iu tr T tnh trn u ngi l 62 USD/ngi [29]. 5 1.1.3. Phn loi Theo phn loi ca ADA nm 2007, T c chia lm 4 nhm [23]: Ttyp1hayTphthucinsulin:lhuqucaqutrnhhy hoicc tbo beta ca o ty,thng dnn thiu htinsulintuyt i, cn s dng insulin ngoi sinh duy tr cn bng chuyn ha. Ttyp2hayTkhngphthucinsulin:dokhnginsulinc quan ch km theo suy gim chc nng t bo beta. Ty trng hp c th, c th 1 trong 2 yu t trn ni tri hoc c hai. T thai k: T ph n mang thai thng khi pht t tun th 24 ca thai k, i khi sm hn. Trong qu trnh mang thai s thay i nng estrogen v progesteron lm tng sn t bo gy ra tng insulin mu ngi m.Hnnatronggiaionnycthngimcngsnxutracc hormon c tc ng i lp vi insulin, vic s dng m to nng lng tng ln. V th, khi c th m khng p ng c nhu cu gia tng v insulin th glucose mu s tng cao v xut hin T. Cc th c bit khc: Khimkhuytchcnngtbobeta(MODY).Glucosemutng lc tr tui, khuyt tt ny c tnh di truyn. Gim hot tnh ca insulin do khim khuyt gen. Gm cc th: -Khng insulin typ A -Hi chng Leprechaunism -Hi chng Rabson Mendenhall -T theo cc t chc m -Cc th khc. Bnh ty ngoi tit: tt c nhng tc ng gy tn thng ln ty c th gy T.Gm:vim ty, ung thty,chn thng, cttyton b, x nang ty, nhim sc t st, x si ty v cc bnh khc. 6 Cc bnh ni tit: -Hi chng Cushing -U t bo tit glucagon -U tit aldosterol -Cc bnh khc. T do thuc hoc ha cht: vacor (thuc dit chut); thiazid; cc cht ng vn chuyn -adrenergic; acid nicotinic; glucocorticoid; Tdonhimtrng:bnhsi(Rubella)bmsinh;Adenovirus; Cytomegalovirus; 1.1.4. Tiu chun chn on TiuchunchnonTtheoLinonTquct(IDF)nm 2010 [29]: -HbA1c 6,5% hoc -Nng glucose mu lc i 7,0 mmol /L ( 126 mg/dL) hoc -Ccctriuchngcatngglucosemuvnngglucosemu thi im bt k 11,1 mmol /L (200 mg/dL) hoc -Nngglucosemuthiimhaigisaunghimphpdungnp glucose ng ung 11,1 mmol/L (200 mg/dL). 1.1.5. Cc xt nghim ha sinh chn on bnh T MtsxtnghimhasinhthngsdngchnonT [10,14]: -Glucose mu lc i -Glucose niu -Xc nh t l HbA1C -Fructosamin trong huyt tng 7 -nh lng peptid C trong mu -Glucose mu ngu nhin -Nghim php tng glucose mu bng ng ung -nh lng insulin. 1.1.6. Cc bin chng ca bnh T1.1.6.1. Bin chng cp tnh H glucose mu Thnggptaibinhglucosemunhngbnhnhndngthuc Tquliuhocdngthucchobnhnhnlci,bba.Ccctriu chng v m hi, chong vng, hoa mt, l m, co git hoc hn m [3,10]. Hn m toan ceton LbinchngcptnhcabnhTcnguyctvongcao.Vic tngcchormongytngglucosemuvsthiuhtinsulinlmtngsn xutglucosetigan,gimthoihaglucose,tngthoihalipidlmtng tnghpthcetongytoanceton.Huqucuicngdntitnhtrngli niu thm thu gy mt nc v in gii, toan chuyn ha mu. Thng gp hn m nhim toan ceton bnh nhn T typ 1 [3,10].Hn m do tng p lc thm thu Bin chng ny hay xy ra bnh nhn T typ 2. Glucose mu tng cao gy tng p lc thm thumu, dn ti tnh trng li tiu thm thu gy mt nc v in gii nng, tt huyt p v hn m [3]. 1.1.6.2. Bin chng mn tnh Bin chng mch mu ln Bin chng mch mu ln, hu qu ca qu trnh x va cc mch mu va v ln,chim ti 80% ccnguynnhn gyt vong bnhnhnT. Bin chng mch mu ln bao gm: bnh ng mch vnh tim, tai bin mch 8 muno,bnhmchmungoivi.Ccyutnguycgm:khng insulin, tng glucose mu, ri lon lipid mu, ht thuc l, bo ph [3]. Bin chng mch mu nh Nhng bin chng ny c lin quan ti tnh trng glucose mu tng cao vcthngnngakhiglucosemuckimsotchtch.Tnthng ch yu cc mao mch v cc tiu ng mch tin mao mch vi biu hin dy mng y, tng tnh thm mao mch v mao mch d v. Hay gp bnh l vi mch mt s c quan: mt, thn, thn kinh [3].1.1.6.3. Cc bin chng khc Nhim trng, bnh l bn chn, tn thng khp [3,10]. 1.1.7. Cc thuc iu tr T Theo ngun gc thuc iu tr T c chia thnh 2 nhm: -Cc thuc tn dc iu tr T -Cc dc liu iu tr T. 1.1.7.1. Cc thuc tn dc Insulin -Ch nh [3,22]:L bt buc i vi T typ 1, T thai k. T typ 2 khi c: tng glucose mu vi tng ceton mu cp nng; can thip ngoi khoa, c thai, suy gan, thn; d ng vi cc thuc vin h glucose mu; ch nh tm thi ngay khi c glucose mu tng cao > 250 - 300mg/dl (14 - 16,5 mmol/l), HbA1c > 11%. T c hn m toan ceton hoc tng p lc thm thu.T do bnh l ty: vim ty mn, sau phu thut ct ty, Trong mt s trng hp nhu cu insulin ca bnh nhn tng cao (iu tr mt s thuc gy tng glucose mu nh corticoid). 9 -Tc dng ph [10,22]: Hglucosemu:ltaibinthnggp,nguynnhnchyuldo dng qu liu insulin. Phn ng d ng vi insulin: xut hin ti ch hoc ton thn. Ni my ay ton thn, chong phn v t gp, c th c mt s triu chng ton thn nh mt mi, st, nga, au cc khp, ri lon tiu ha. Lon dng m m ti ch tim: teo m m di da Cc sulfonylure -Chnh:Ttyp2thtrngtrungbnhhocgy,phihpvi metformin, thiazolidindion, acarbose, insulin [3,10,22]. -Tc dng ph: h glucosemu, d ng ngoi da, da, nga, nimy ay,damncmvinhsng,vngda;rilontiuha,bunnn,nn; gim bch cu, gim tiu cu [3,10,22]. Cc biguanid -Ch nh: T typ 2, nht l bnh nhn c tha cn hay bo ph [3,34]. -Tc dng ph: cc tc dng ph trn ng tiu ha nh chn n, bun nn, nn, y bng, tiu chy, gp 20% bnh nhn. Tc dng ph ny c lin quan n liu lng hay xy ra khi bt u iu tr, c 3 - 5% bnh nhn phingngthuc.Gynhimtoanlactickhisdngchongicaotui, nghin ru, suy gan - thn, suy tim hoc suy h hp [3,22]. Cc thiazolidindion -Chnh:kthpvisulfonylurehocmetforminhocinsulintrong iu tr T typ 2 [3]. -Tcdngph:thnggytngcn,chyudolmtngtchtrm didavmtphndoginc.Vvy,cnthntrngkhiiutr thiazolidindioncho ccbnh nhn suytim hoccbnh tim,vim ganhoc men gan cao [3]. 10 Thuc c ch enzym - glucosidase lm gim hp thu glucose -Ch nh: tng nh glucose mu sau n, iu tr n tr liu kt hp vi ch n hoc phi hp vi cc thuc khc [3]. -Tc dng ph: bun nn, y trng bng; cm gic mt i ngoi, tiu chy [3]. 1.1.7.2. Cc dc liu iu tr T CcthuchadcchiuquiutrTcao.Tuynhin,cn nhng hn ch nh c nhiu tc dng ph, chi ph iu tr cao v ngi bnh c xu hng phi tng liu iu tr sau mt thi gian di dng thuc. Hn na, Tlmtbnhmntnhcniutrtrongthigianludi.Dohin nay, s dng cc dc liu iu tr T vi u im gi thnh r, t tc dng ph l hng la chn hp l gii quyt vn ny. Cc nghin cu trong v ngoincchothycnhiudcliuchotc dnghglucosemutt c s dng trong iu tr T. Mt s dc liu nh: -Dy tha canh (Gymnema sylvestre) B phn dng: l. Hot cht tc dng l acid gymnemic [16]. -C ngt (Stevia rebaudiana) B phn dng: phn trn mt t. Hot cht l steviosid, kch thch bi tit insulin t cc t bo o ty Langerhans, iu tr T typ 2 [12]. -Sinh a (Rehmania glutinosa) B phn dng: c sy kh c v ngt ng, tnh hn. Hot cht gy tc dng h glucose mu: catalpol v mannozid [12]. -Bng i (Lantana camara) Cao nc l bng i lm gim glucose mu ca chut cng b gy T bng alloxan [12].11 -Mp ng (Momordica charantia) Nhiunghincucho thydchchitquvthnmp ngvicc dungmikhcnhauuctcdnghglucosemutrnccmhnhgy tng glucose mu thc nghim. Hot cht l glycosid [9,17]. VitNamsnxutthnhcngvinnangMorantinvihotcht chnh l glycosid v th nghim lm sng t kt qu tt trn ngi [17]. -Bng lng nc (Lagerstroemia speciosa) Dchchitlbnglngncctcdnghglucosemucachut nht b gy T bng STZ [8]. Ngoira,mtscykhcvangcnghincutcdngh glucose mu nh: th phc linh, hong k, huyn sm, dip h chu, d, [5,8,11,12,17]. 1.2.Tng quan v chi Gymnema R.Br. 1.2.1. V tr phn loi V tr ca chi Gymnema R.Br. trong h thng phn loi Takhtajan cng b nm 2009, chi Gymnema R.Br. c v tr phn loi nh sau [45]: Ngnh Ngc Lan (Magnoliophyta) Lp Ngc Lan (Magnoliopsida) Phn lp Bc H (Lamiidae) B Long m (Gentianales) H Trc o (Apocynaceae) Phn h Thin L (Asclepiadaceae) Chi Gymnema R.Br. 1.2.2. c im thc vt v phn b 1.2.2.1. c im chung ca chi Gymnema R.Br. Cyleo,khng c rphtrnthn.Lmc i, khngnc.Cmhoa xim, dng tn n chm. Hoa nh. Thyi nh, hnh trng, u t, gc i 12 ctuyn.Trnghnhbnhxe;thytrngkhnggptrongn,tinkhaivn phi. Trng ph n, vy trng ph nh trng, thng c cc hng lng xp dc trng. Ch nh dnh nhau, bao phn 2 , ht phn dnh thnh khi phn; c quantruynphncgtdnhv2chui,khiphnhngln,chcmt khi phn trong mi phn. u nhy phnh ln hnh trng, nh bu khng tht li thnh dng vi nhy. Ct nh - nhy hnh ng nhn u [1,7,33]. Phnb:chiGymnemaR.Br.ckhong25loi,phnbvngTy ChuPhi,Australia,chu.VitNamc4loilGymnemainodorum (Lour.)Decne;GymnemalatifoliumWall.ExWight;Gymnemayunnanense Tsiang; v Gymnema sylvestre (Retz) R.Br. ex Schul [1]. 1.2.2.2. Loi Gymnema sylvestre (Retz) R.Br. ex Schul Dyleo,cao6-10m,toncycnhamvng.Thnmanglc lng, c l b tha. L c phin bu dc, trng ngc, di 6 - 7 cm, rng 2,5 - 5 cm, u nhn, gn bn 4 - 6 i, r mt di, nhn lc kh, cung di 5 - 8 mm. Hoa nh, mu vng, xp thnh xim dng tn nch l, cao 8 mm. i clngmnvralng.Trngkhngclngmtngoi,trngphc5 rng. Qu i di 5,5 cm, rng na di. Ht dp, mo lng di 3 cm [4,35]. Phn b: n , Vit Nam, Nam Trung Quc, Indonesia. Vit Nam, phnbkhpcnc:minbccThiNguyn,Namnh,VnhPhc, min trung c Qung Tr, Thanh Ha [4]. 1.2.2.3. Loi Gymnema latifolium Wall. Ex Wight Cy leo n 10m. Thn nhn, cnh hoa mu ti xm, c lng. Phin l rng hnh trng thun n hnh trng hay trng rng, nhn hay c lng mnh dctheogn;nhnhncui;gctrnnhnhtimrng,gnbn4-6 cp, cung l di 2 - 6 cm. Cm hoa xim, cao n 4 cm; cung hoa di 1 - 1,5 cm. Thy i hnh thun, ngn hn ng trng, c lng. Trng mu vng, cao 6 -7mm,mtngoiclng;ngtrnghnhtr, nhn,thytrnghnh thun, 13 nh trn, nhn ngoi tr mp c lng. Trng ph n, vy trng ph dnh trng, c hng lng xp dc theo trng ph. Khi phn thun. u nhy dng vm, vt cao hn ng trng. Qu i, hnh mc, vch dy v hi c si. Ht c khong 1,51 cm, mo lng di 4 cm [1,18].Phn b: trn th gii phn b n , Trung Quc, Vit Nam, Lo, Indonesia, Philippin. Vit Nam, c nhiu Thi Nguyn, Ha Bnh,Tuyn Quang, Nam B [1,7]. 1.2.2.4. Loi Gymnema inodorum (Lour.) Decne Dyleoti10m.Thnnhn,ccnhnhnonmununht,clv, ph lng mng. Cung l 2 - 6 cm, l hnh trng thun hoc hnh trng rng, nhnhocph lng mng dc gnl. Cm hoa ximdngchmxp ging nh u, xp xon c ti 4 cm. i thun c lng mng hoc lng mn. Trng hoa mu vng, 6 - 7 mm, c lng mn dy c pha ngoi ng trng dng hnh tr.Khi phn hnh ch nht.u nmnhyc hnhmivm, thln trn. Qu 2 i, hnh mc. V qu nhn. Ht khong 1,5 1 cm, mo lng khong 4 cm [18].Phn b: Trung Quc, Thi Lan, Vit Nam, n . Vit Nam phn b Nam B, Nha Trang, Bc Kn, Thi Nguyn, Ha Bnh [1,7]. 1.2.2.5. Loi Gymnema yunnanense Tsiang Thn bi leo, cao ti 8 m. Thn gi phnh ln thnh cc rnh su, thn non c nhiu l v, b mt c ph lng dy cmu vng st. Ton thn c nhamvng. Li, phin l hnhtrng,bu dc hoctrngngc, 5- 6 cp gn lng chim. Ton b b mt l v gn l c ph lng st nu. Cm hoa ximmci xngnchl.L iri, hnhtrng,clngdyc.Trng hnlin,muvng.ngthyhnhtrng,cphnphhngtrngmang lngdymuvngnu,phnphcanhrng,thcaolntrnngtrng nhng vn thp hn u nm nhy. Khi phn hnh ch nht, thng ng. B 14 nhy gm 2 l non ri nhau. Ht hnh trng thun di, mo lng 2,5 cm [18].Phn b: Vit Nam, Trung Quc. Vit Nam, phn b ch yu Ty Nguyn, Kin Giang [1]. 1.2.3. Thnh phn ha hc TronglccloithucchiGymnemaR.Br.calbumin,carbohydrat, inositol,acidtartaric,acidformic,acidhuc,cchpchtanthraquinolic, ng kh, acid amin, nha, saponin, coumarin, tanin, flavonoid [4,6]. Thnh phn chnh c tc dng h glucose mu c xc nh l nhm dnchtacidgymnemic,mtacidhucthucnhmsaponintriterpenoid [16,19,39].Nhiu nghin cu xc nh cu trc acid gymnemic ca G.sylvestreNm1967v1969,Stocklinvcngsxcnhc gymnemageninl saponin chnhctrongl G.sylvestrevl sn phmthy phncaacidgymnemic,ccutrcl3,16,21,22,23,28- hexahydroxyolean - 12 - en [39].Nm1989v1992,Yoshikawalnltcngbphnlpc12 acid gymnemic I XII [39].Nm 2004, Liu X. v cng s xc nh c trong l G.sylvestre c kaempferol(3ODglucopyranosyl-(14)L rhamnopyranosyl-(16)D-galactopyranoid)vquercetin(3O- 6- (3- hydroxyl- 3- methylglutaryl)- - D- glucopyranoside) [31]. Nm 2008, Zhu v cng s phn lp c 2 loi saponin triterpenoid tlG.sylvestrel16-hydroxylolean12en3O-[D-glucopyranosyl(16)D-glucopyranosyl]28OD- glucopyranosidev16,21,28trihydroxylolean12ene3O- glucoronopyranoside [49]. 15 Gurmarinmtpolypeptidckhnnglmmtcmgicngtm khng nh hng ti cc v gic khc, c phn lp t l G.sylvestre [46].Nm 2001, Kazumasa Shimizu v cng s phn lp c 4 loi acid gymnemic trong l G.inodorum [43]: GiA-1 l acid (3,16,22) - 16,28 dihydroxyolean 12 en 3 yl O D - glucopyranosyl D - glucopyranosiduronic.GiA-2 l acid (3,4,16) - 16,23,28 trihydroxyolean 12 en 3 yl - D - glucopyranosiduronic.GiA-5lacid (3,4,16,22) 22 -(N -methylanthraniloxy) -16,23,28- trihidoroxyolean12en3yl3ODglucopyranosylD- glucopyranosiduronic GiA-7 l acid (3,4,16,22) 22 - (N methylanthranil - oxy) - 16,23,28 trihydroxyolean 12 en 3 yl D glucopyra - nosiduronic. Ngoi nhng thnh phn c bn, hin cha c nghin cu xc nh cu trc c th ca acid gymnemic c trng G.latifolium v G.yunnanense. 1.2.4. Tc dng iu tr T c nhiu nghin cu ch ra tc dng h glucose mu trn in vitro v in vivo ca chi Gymnema R.Br. [21,38]. Trong phn ln cc nghin cu l v loi G.sylvestre, cc loi khc c rt t kt qu nghin cu c cng b. Tronghuhtcctiliuthamkhovtcdngsinhhcca G.sylvestreucnintcdnghglucosemu[4,6,16,21,27,40],tc dng ny c chng minh qua nhiu m hnh thc nghim v trn lm sng. Trong y hc c truyn n , bt l G.sylvestre c s dng liu 6 - 60 g/ngy c tc dng h glucose mu tt [41]. Th nghim lm sng trn bnh nhnTtypII:cho22bnhnhnungcaoGymnemasylvestreliu400 mg/ngy, trong 18 - 20 thng kt hp thuc iu tr tiu ng. Nhm iu tr c s gim glucose mu ng k v tng lng insulin tit ra t ty [24]. 16 Theo mt nghin cu ca Sugihara Y. v cng s, acid gymnemic liu 13,4mg/kglmgim14,060,0%lngngtrongvng6gisovi glibenclamid; c ch c xc nh l lm tng nng insulin huyt tng chutnhtTmkhngcchhotngcaglucosidase[46].M hnhthcnghimcaPrabhuS.,VijayakumarS.trnchutbtngglucose mubngSTZchoktqu:ungdchchitMeOHcalG.sylvestremi ngy vi cc liu 100, 200 v 400 mg/kg trng lng c th u cho tc dng h glucosemusovi nhmchng(p 26 kg/m2. Cc i tng c n vi ch 2000 kcal/ngy kt hp vi dch chit l G.sylvestre v c s gim st v hot ng thlc.Sau tunth8kt qucho thytrnglngc th v BMIgimt5-6%;tnglngcholesterol,LDLvcctrigliceridtrong huyt thanh u gim khi lng HDL trongmu v s bi tit cht bo tng ln [41]. Hot tnh khng khun Trong nghin cu in vitro, dch chit EtOH ca l G.sylvestre cho thy hottnhkhngkhunviccchngvikhunBacilluspuntilis,Bacillus subtilis,Pseudomonasaeruginosa v Staphylococusaureusmkhngctc dng vi Escherichia coli v Proteus vulgaris [41,46]. Tc dng lm mt cm gic ngt Tc dng ny do gurmarin, polypeptid phn lp c t G.sylvetre gy ra.Nlmcchchnlccmgicngtmkhngnhhngnccv gic khc chut cng . Tc dng mt cm gic ngt ko di kh lu 2 - 3h, tcdng ny smti nhanhchng ditcdngcachtkhng gurmarin trong huyt tng [28,46]. 18 CHNG 2. NGUYN LIU, I TNG, PHNG TIN V PHNG PHP NGHIN CU 2.1.Nguyn liu Nguynliullcaccmuthuc4loitrongchiGymnemaR.Br. c thu hi cc a phng khc nhau. Ccmu thu v u c phi se trong bng rm, sy kh 45 50oC, xay nh thnh bt, hm m khng qu 5%. Thng tin v danh php, ni thu mu v m k hiu ca cc mu s dng trong nghin cu c trnh by trong bng 2.1. Bng 2.1. Cc mu s dng trong nghin cu SttLoiM muNi thu mu 1. Gymnema sylvestre(Retz.) R.Br. ex Schult S.QNQung Ninh 2.S.PYPh Yn 3.S.VPVnh Phc 4.S.NNam nh 5.S.QTQung Tr 6.S.ThNThi Nguyn 7. Gymnema latifoliumWall. ex Wight L.ThNThi Nguyn 8.L.HBHa Bnh 9.L.TQTuyn Quang 10.L.GLGia Lai 11.L.TNTy Ninh 12. Gymnema inodorum(Lour.) Decne I.HNH Ni 13.I.BKBc Kn 14.I.VPVnh Phc 15. Gymnema yunnanense Tsiang Y.GLGia Lai 16.Y.KTKon Tum 17.Y.L k Lk 19 2.2.i tng nghin cu Chut nht trng chng Swiss khe mnh, ging c, trng lng trung bnh 27 2g, mua ti hc vin Qun y, chut c chia thnh cc l, mi l 7 - 10 con. 2.3.Phng tin nghin cu 2.3.1. Ha cht -Streptozotocin (SIGMA)-Gliclazid 80mg (STADA) -Cc ha cht khc t tiu chun phng th nghim. 2.3.2. Thit b dng chit v pha ch dch chit -B bnh chit ngm kit -My ct quay -Cc c m, a thy tinh, ng ong, 2.3.3. Thit b dng trong qu trnh th tc dng sinh hc -My o glucose mu ACCU CHEK ACTIVE (Roche) -Que th o glucose mu (Roche) 2.4.Phng php nghin cu 2.4.1. Phng php chit xut dc liu Dcliucsykh45-50oC,xaynhthnhbt,chitbng phng php ngm kit vi dung mi l ethanol 80o, ngm dc liu trong 48 gi, sau rt dch chit vi tc 1 ml/pht, m bo cn lun ngp dc liu trong qu trnh rt, p kit.Dch chit thu c ct thu hi dung mi di p sut gim cho ti cn, sau pha hn dch cn vi nc ct t l 1:1, s dng NaCMC 0,5% n nh hn dch. 20 2.4.2. Phng php nh lng glucose huyt Phng php enzym glucose oxidase vi my t ng v que th Accu chek Active. Chut c ly mu ui, b git mu u, ri nh vo que th cm sn trong my. Nh xc tc ca enzym glucose oxidase (GOD), glucose c oxi ha thnh acid gluconic v H2O2 theo phn ng: Glucose + H2O + O2 Acid gluconic + H2O2 H2O2 to thnh s b peroxidase phn hy, gii phng O2, oxy ha O - dianisidin to thnh phc cht c mu vng nu theo phn ng: O- dianisidin + H2O2 phc hp mu vng nu + H2O Cngmucxcnhbngphngphpoquangtlvi lng glucose trong mu nh lng. 2.4.3. Phng php gy tng glucose mu bng streptozocin Streptozocin(STZ)lmtkhngsinhphrng,chitxuttnm Streptomyces achromogennes. Ging vi cc tc nhn alkyl ha khc, n gy ph hy cu trc si ADN ca t bo . Mt khc n c i lc cao vi knh vn chuyn glucose GLUT2 c mt nhiu mng t bo , do gy c c hiu cho t bo , ph hy t bo ny, do gy thiu ht insulin v gy tng glucose mu. Mc tn thng tiu o Langerhans ty thuc vo liu STZ v chng ng vt, c th gy tn thng mt phn hoc hon ton cc t bo ca o ty [16,26]. gy tng glucose mu thc nghim, cc nghin cu thng s dng liu STZ dao ng t 40 - 300 mg/kg.TrnchutnhttrngthcnghimchngSwiss,liuSTZ150mg/kg timmngbngcnhiutcgilachngytngglucosemuthc nghim [11,16,19]. STZ c pha trong m natri citrat pH 4,5 nng 0,1M [25]. 21 2.4.4. Phng php nghin cu tc dng h glucose mu trn ng vt th nghim S dng phng php nghin cu thc nghim c i chng. nh gi tcdnghglucosemucadchchitdcliutrnmhnhchuttng glucosemuthcnghimbngSTZ(timmngbngliu150mg/kg).Kt qunghincuctinhnhsosnhgianhmthvnhmichng trong cng iu kin. 72hsaukhitimmngbngdungdchSTZ(150mg/kg),tinhnh nhlngglucosemulcicachut.Lachnchutcglucosemu 10 mmol/L cho th nghim.Chut tng glucose mu thc nghim c chia thnh cc l:-L chng trng ung dung dch NaCMC 0,5% -L chng dng ung gliclazid (20 mg/kg) -Cc l th ung hn dch chit t dc liu Chochutungmuthtrongvng7ngy(vo8hsngmingy), trong 7 ngy cho chut n ung bnh thng. Sau ln ung cui cng, nh lngglucosemulcicachut.Sosnhsthayiglucosemuca cc l th nghim. 2.4.5. X l s liu S liu c x l bng xc sut thng k vi Test student v c h tr bi phn mm Microsoft Excel 2007. S khc bit c ngha thng k khi p < 0,05. Biu din s liu:X SE. 22 CHNG 3. THC NGHIM, KT QU V BN LUN 3.1.Kt qu 3.1.1. Xcnhliudngthchhpctcdnghglucosemutrnm hnh chut b gy tng glucose mu bi STZ liu 150 mg/kg Trc khitin hnh nh gi v sosnh tc dng h glucose mu ca cc mu Gymnema sp., vn t ra l cn phi la chn mt liu dng thch hp. V vy, chng ti tin hnh nghin cu s ph thuc gia tc dng v liu lng ca mu th vi 3 liu khc nhau trn m hnh chut tng glucose mu thc nghim bi STZ 150 mg/kg. Thc nghim c tin hnh c tin hnh vi 1 mu Gymnema sylvestre (Retz.) R.Br. ex Schult, loi Gymnema c khng nh v tc dng h glucose mu.Chut tng glucose mu thc nghim c chia thnh 5 l: -L chng trng ung dung dch NaCMC 0,5% -L chng dng ung gliclazid 20 mg/kg -L th ung hn dch cn dc liu liu 5 g/kg -L th ung hn dch cn dc liu liu 10 g/kg -L th ung hn dch cn dc liu liu 15 g/kg (Liu tnh theo gam dc liu kh/ kg th trng chut). Cho chut ung trong vng 7 ngy.nhlngglucosemutrcvsautiutr.Ktquthhin bng 3.1 v hnh 3.1. 23 Bng 3.1. S ph thuc liu dng G.sylvestre (Retz.) R.Br. ex Schult v tc dng h glucose mu trn m hnh chut tng glucose mu thc nghim bi STZ L Glucose mu (mmol/l) Mc h glucose mu (%) p so vi l Ngy 0Ngy 71234 1 Chng trng 21,7 2,520,6 2,24,05 2,18 2 Chng dng 20,9 2,311,8 1,343,47 1,50p2,1 < 0,001 3 Th5g/kg 20,7 2,515,1 2,328,04 3,24p3,1 < 0,05p3,2 < 0,01 4 Th 10g/kg 20,4 2,111,8 1,442,16 1,49p4,1 < 0,001p4,2 > 0,05p4,3 < 0,05 5 Th 15g/kg 21,5 1,811,3 1,047,41 2,54p5,1 < 0,001p5,2 > 0,05p5,3 < 0,05p5,4 > 0,05 24 Hnh 3.1. nh hng ca liu dng G.sylvestre (Retz.) R.Br. ex Schult n mc h glucose mu trn chut tng glucose mu thc nghim bi STZ Nhn xt: -Tt c cc l th vi liu dng khc nhau v l chng dng u c tc dng h glucose mu khc bit c ngha vi l chng trng. -So snh cc l th vi l chng dng: l th liu 5 g/kg tc dng thp hnlchngdngcngha(p 0,05). -Trong3lth,lthliu15g/kgchotcdngcaonht(47,41%) tngng vitc dnglthliu10 g/kg(p>0,05).L thliu5g/kg cho tc dng thp nht (28,04%) v khc bit vi 2 l th cn li (p < 0,05). 25 T cc kt qu trn, liu 10 g/kg (tnh theo gam dc liu kh/ kg th trngchut)clachnlliudngthchhpcs dngchoccth nghim tip theo. 3.1.2. TcdnghglucosemucaGymnemasylvestre(Retz.)R.Br.ex Schult nh gi tc dng h glucosemu ca cc loi Gymnema sylvestre (Retz.)R.Br.exSchult, tinhnhthcnghimcichngtrnchuttng glucose mu thc nghim bng STZ. Chia chut thnh 8 l: -1 l chng trng (ung dung dch NaCMC 0,5%) -1 l chng dng (ung gliclazid 20 mg/kg)-6lth(unghndchcndcliuviliu10g/kgca6muG. sylvestrethuc6vng:QungNinh(S.QN),PhYn(S.PY),Vnh Phc (S.VP), Nam nh (S.N), Qung Tr (S.QT), Thi Nguyn (S.ThN)). Chut ung mu th trong vng 7 ngy. So snh s thay i glucosemu ca cc l sau thi gian iu tr. Kt qu th hin bng 3.2 v hnh 3.2. 26 Bng 3.2. S thay i glucose mu ca cc l chut ungG.sylvestre (Retz.) R.Br. ex Schult L Glucose mu (mmol/l) T l h glucose mu (%) p so vi l Ngy 0Ngy 712 1 Chng trng 17,5 1,716,8 1,7 3,47 1,81 2 Chng dng 17,7 1,910,1 0,8 41,75 2,88 p2,1 < 0,001 3S.QN16,9 1,210,8 0,9 35,97 2,46 p3,1 < 0,001p3,2 > 0,05 4S.PY16,5 0,89,6 0,9 42,24 3,09 p4,1 < 0,001p4,2 > 0,05 5S.VP16,8 1,411,8 1,0 29,94 2,08 p5,1 < 0,01p5,2 < 0,05 6S.N16,8 0,811,3 0,6 32,42 2,20 p6,1 < 0,01p6,2 < 0,05 7S.QT16,8 0,612,6 0,6 25,19 2,30 p7,1 < 0,05p7,2 < 0,01 8S.ThN17,7 1,610,8 0,8 38,46 2,08 p8,1 < 0,001p8,2 > 0,05 27 Hnh 3.2. T l % h glucose mu ca cc l chut ungG.sylvestre (Retz.) R.Br. ex Schult Nhn xt: -CcmuG.sylvestreuctcdnghglucosemukhcbitc ngh thng k so vi l chng trng. -Khi so snh cc l th vi l chng dng, kt qu l mc h glucose muca3lthviccmuG.sylvestreQungNinh,PhYn,Thi Nguyn khng khc bit vi mc h ca l chng dng (p > 0,05). Ba mu cn li cho tc dng km hn gliclazid 20 mg/kg (p < 0,05). -Trong cc mu G. sylvestre, mu Ph Yn cho t l h glucose mu cao nht (42,24%), mu Qung Tr cho mc h glucose mu thp nht (25,19%). 3.1.3. Tc dng h glucose mu ca Gymnema latifolium Wall. ex Wight Chiachutthnh7ltrong1lchngtrng(ungdungdch NaCMC 0,5%), 1 l chng dng ung gliclazid 20 mg/kg v 5 l th (ung 28 hn dch cn dc liu vi liu 10 g/kg ca 5 mu G.latifolium thu c 5 vng:ThiNguyn(L.ThN),HaBnh(L.HB),TuynQuang(L.TQ),Gia Lai (L.GL), Ty Ninh (L.TN)). Chut ung mu th trong vng 7 ngy. Sosnhsthayiglucosemucachutsau thigian iutr.Kt qu th hin bng 3.3 v hnh 3.3. Bng 3.3. S thay i glucose mu ca cc l chut ungG.latifolium Wall. ex Wight L Glucose mu (mmol/l)T l h glucose mu (%) p so vi l Ngy 0Ngy 712 1 Chng trng 17,5 1,716,8 1,7 3,47 1,81 2 Chng dng 17,4 1,49,8 0,7 43,17 1,58 p2,1 < 0,001 3L.ThN16,7 1,310,3 1,0 38,25 4,10 p3,1 < 0,001p3,2 > 0,05 4L.HB16,1 1,29,0 0,9 44,24 1,04 p4,1 < 0,001p4,2 > 0,05 5L.TQ16,6 1,310,2 1,0 39,09 2,60 p5,1 < 0,001p5,2 > 0,05 6L.GL16,8 1,09,9 0,7 41,04 2,14 p6,1 < 0,001p6,2 > 0,05 7L.TN16,3 0,710,3 0,5 36,92 2,21 p7,1 < 0,001p7,2 < 0,05 29 Hnh 3.3. T l % h glucose mu ca cc l chut ungG.latifolium Wall. ex Wight Nhn xt: -5muG.latifoliumulmgimglucosemukhcbitcngha thng k so vi l trng (p < 0,001).-Khisosnhcclthvilchngdng,chclthmu G.latifolium Ty Ninh c s khc bit c ngha thng k (p < 0,05), cc l mu th cn li so vi l chng dng s khc bit khng c ngha thng k (p > 0,05). -Trong cc mu G.latifolium, mu Ha Bnh c mc h glucose mu cao nht (44,24%). 3.1.4. Tc dng h glucose mu ca Gymnema inodorum (Lour.) Decne Tinhnhthnghimvi1lchngtrngungdungdchNaCMC 0,5%,1lchng dngunggliclazid 20mg/kgv3l thunghndch 30 cn dc liu liu 10 g/kg ca 3 mu Gymnema inodorum: Vnh Phc (I.VP), H Ni (I.HN), Bc Kn (I.BK). Chut ung mu th trong vng 7 ngy.So snh s thay i glucose mu sau thi gian th nghim. Kt qu thu c th hin bng 3.4 v hnh 3.4. Bng 3.4. S thay i glucose mu ca cc l chut ungG.inodorum (Lour.) Decne L Glucose mu (mmol/l) T l h glucose mu (%) p so vi l Ngy 0Ngy 712 1 Chng trng 13,9 1,014,2 1,1 -1,89 1,80 2 Chng dng 16,0 1,69,2 0,7 41,33 2,75 p2,1 < 0,001 3I.VP14,0 1,710,0 0,7 27,00 2,70 p3,1 < 0,05p3,2 < 0,05 4I.HN14,3 1,710,5 0,9 32,82 2,32 p4,1 < 0,05p4,2 < 0,05 5I.BK14,3 1,210,5 0,7 25,72 3,26 p5,1 < 0,05p5,2 < 0,05 31 Hnh 3.4. T l % h glucose mu ca cc l chut ungG.inodorum (Lour.) Decne Nhn xt: -CcmuG.inodorumuctcdnghglucosemukhcbitc ngha thng k so vi l trng (p < 0,05) -TcdnghglucosemucaonhtlcamuG.inodorumHNi (32,82%). -Sovilchngdng,mchglucosemucacc lmuththp hn c ngha so vi l chng dng (p < 0,05). 3.1.5. Tc dng h glucose mu ca Gymnema yunnanense Tsiang Chiachutthnh5lvi1lchngtrng(ungdungdchNaCMC 0,5%), 1 l chng dng (ung gliclazid liu 20 mg/kg) v 3 l th (ung hn dchcndcliuviliu10g/kgca3muG.yunnanenseKonTum (Y.KT), Gia Lai (Y.GL) v k Lk (Y.L)).32 Cho chut ung mu th trong 7 ngy.Sosnhsthayiglucosemucaccl sauthi gianiutr, kt qu thu c th hin bng 3.5 v hnh 3.5. Bng 3.5. S thay i glucose mu ca cc l chut ungG.yunnanense Tsiang L Glucose mu (mmol/l)T l h glucose mu (%) p so vi l Ngy 0Ngy 712 1 Chng trng 17,5 1,716,9 1,6 2,73 2,88 2 Chng dng 14,6 1,48,4 0,5 41,23 2,75 p2,1 < 0,001 3Y.KT16,9 1,613,8 1,0 22,03 2,43 p3,1 < 0,05p3,2 < 0,001 4Y.GL14,0 1,111,3 0,6 18,86 2,67 p4,1 < 0,05p4,2< 0,001 5Y.L16,4 1,413,3 1,3 19,39 1,55 p5,1 < 0,05p5,2 < 0,001 33 Hnh 3.5. T l % h glucose mu ca cc l chut ungG.yunnanense Tsiang Nhn xt:-CcmuG.yunnanensectcdnghglucosemukhcbitc ngha thng k so vi l trng (p < 0,05). -MuG.yunnanenseKonTumchotlhglucosemucaonht (22,03%). -Tlhglucosemucacclththphnvkhcbitsovil chng dng (p < 0,001). 3.1.6. So snh mc h glucose mu gia cc loi Gymnema R.Br. c th so snh mt cch khi qut tc dng h glucose mu gia cc loi Gymnema R.Br. vi nhau, vi mi loi chn mu cho tc dng tt nht so snh. Kt qu c th hin hnh 3.6. 34 Hnh 3.6. T l % h glucose mu ca cc loi Gymnema R.Br. Nhn xt:Trong4loi,loiG.latifoliumchotcdnghglucosemuttnht (44,24%),loiG.yunnanensechotcdngthpnht(22,03%).G.sylvestre cng cho tc dng tng i tt (42,24%). 3.2.Bn lun 3.2.1. TcdnghglucosemucaGymnemasylvestre(Retz.)R.Br.ex Schult Viliu10g/kg(tnhtheogamdcliukh),ttcccmu G.sylvestre thu c u lm h glucose mu c ngha trn m hnh chut tngglucosemuthcnghimbiSTZ.iunykhngnhGymnema sylvestre Vit Nam cng c tc dng h glucose mu ging nh cc nghin cu v G.sylvestre cng b trn th gii [24,27,37,38,40,41]. TrongccmuthucthmuG.sylvestrePhYnchomch glucosemucaonht(42,24%),2muG.sylvestreQungNinhvThi Nguyn cng cho tc dng tt vi mc h glucose mu ln lt l 35,97% v 35 38,46%. C 3 mu u c tc dng tng ng vi gliclazid liu 20 mg/kg. CcmunycngchoktqutthnktqumTrngThTm(2008) cng b trn mu G.sylvestre Thanh Ha (32,21%) vi cng liu dng v m hnhthcnghim[16].ngthi,khisosnhviktquthcnghimca Sathya S. v cng s (2008) th mc h glucose mu ca mu G.sylvestre Ph Yn (42,24%) ln hnmc h glucosemu ca G.sylvestrem Sathya S. v cngscngb(39,97%)[40].G.sylvestrelmtloidcliuv ang c s dng rng ri trong thc t iu tr T. Do vy cc kt qu ny rt c ngha trong vic la chn m rng cc vng nguyn liu hin c.S khc nhau v mc h glucose mu ca cc mu G.sylvestre thu hi cc vng a l khc nhau c th do chng c ngun gen khc nhau, hoc chu nh hng bi cc iu kin kh hu, th nhng khc nhau. Cn c thm cc nghin cu v cc yu t nh hng ny nhm thu c nhng ngun nguyn liu Gymnema sylvestre cho cht lng v sn lng cao. 3.2.2. Tc dng h glucose mu ca Gymnema latifolium Wall. ex Wight Cc mu G.latifolium vi liu 10 g/kg u c tc dng h glucose mu trnmhnhchuttngglucosemuthcnghimbiSTZ.Trong,mu G.latifolium Ha Bnh cho mc h glucose mu cao nht (44,24%) cng vi ba mu Thi Nguyn (38,25%), Tuyn Quang (39,09%), Gia Lai (41,04%) tngngvigliclazid20mg/kg,muTyNinhtcdngthpnht (36,92%).KtqunyhontonphhpviktqumPhmVnHi (2010) tin hnh th nghim trn m hnh tng t, mu G.latifolium thu Ha Bnh cho mc h glucose mu sau 10 ngy s dng l 40,84% [6].Gymnemalatifoliumvitnthnggildythacanhltocc im thc vt ng ch l lng sinh khi l ln hn nhiu so vi dy tha canh(Gymnemasylvestre).Trongkhithnhphnctc dnghglucose mu li tp trung phn ln l [39], v th vi kt qu cho thy tc dng h 36 glucosemuttthG.latifoliumhontoncthtrthnhmtngundc liu quan trng iu tr T bn cnh G.sylvestre. Hn na, hin nay, ngoi Vit Nam, trn th gii hu nh cha c cng b v tc dng h glucose mu caG.latifoliumccngb.Dovy,ktqunycnggp nhtnh, gp phn m ra nhng trin vng khai thc ngun nguyn liu ny iu tr T. 3.2.3. Tc dng h glucose mu ca Gymnema inodorum (Lour.) Decne Trnm hnh chut tng glucose mu bi STZ, c 3mu G.inodorum uctcdnghglucosemu.Ktquphhpvinghincuca ShimizuK.chngminhG.inodorumctcdnghglucosemu.Theo ShimizuK.vcngs,dchchitG.inodorumlmgim60%khnnghp thu glucose trn chut cng sau 30 pht [44].TuyloiG.inodorumchacnghincunhiuvtcdngh glucose mu, nhng vi mc gy h glucose mu cao nht l 32,82% ca mu G.inodorumHNithncnglmtngunnguynliutimnngtrong iutrT.Cn tin hnhthmccnghincu c th nh giy hn tc dng ca loi ny. 3.2.4. Tc dng h glucose mu ca Gymnema yunnanense Tsiang Kt qu thc nghim vi mu G.yunnanense Kon Tum cho tc dng tt nht(22,03%)soviccmutrongloi.Ktquchngminhtcdngh nghuytcaloiG.yunanensetrnthcnghim,cngcthmchokt qunghincucaJing-TianXievcngstrnchuttngglucosemu thcnghim,nhmchutsdngdchchitG.yunnanensechotcdngh glucose mu cao hn 22% so vi nhm i chng [48]. Mc d t l h glucose mu khng cao nh cc loi cn li trong chi, nhng vic nghin cu tc dng h glucose mu ca G.yunnanense va cung cpthmmtdcliucthdngtrongiutrT,vagipnhgi 37 khi qut v tc dng ca chi Gymnema R.Br. Vit Nam. T c ci nhn tng qut hn cho cc nghin cu tip theo v chi Gymnema R.Br. 3.2.5. So snh tc dng h glucose mu gia cc loi Gymnema R.Br. so snh tc dng gia cc loi Gymnema vi nhau, vi mi t th ccmu th c sdng vicngmtliu (10 gam dcliukh/ kg th trng),vcngcsosnhvilchngtrngvlchngdng(ung gliclazid 20 mg/kg). Kt qu cho thy: -G.sylvestrevG.latifoliumchotcdnghglucosemutthn G.inodorum v G.yunnanense (hnh 3.6).-Ccmu th ca 2 loi Gymnema inodorum v Gymnema yunnanense cho tc dng thp hn v khc bit c ngha so vi tc dng ca gliclazid. -2loiGymnemasylvestrevGymnemalatifoliumcccmuchotc dngtngngvigliclazid.CcmugmG.sylvestrePhYn, QungNinh,ThiNguynvG.latifoliumThiNguyn,HaBnh,Tuyn Quang, Gia Lai. Cc mu cn li ca 2 loi ny cng cho tc dng nhng thp hn ng k so vi chng dng. T kt qu so snh trn c th gip la chn mt s mu Gymnema c tc dng h glucose mu u th trong cc mu c th nghim, l cc mu: G.sylvestre (Ph Yn, Qung Ninh, Thi Nguyn) v G.latifolium (Thi Nguyn,HaBnh,TuynQuang,GiaLai).Hinnay,trnthtrngc nhiusnphmthucvthcphmchcnnghtriutrT.Tuy nhin,vickhaithcnguynliuchoccsnphmnychacmtcs khoa hcm chyu da vo tnh thun tin ca ni sn xut, nn hiu qu thc t ca cc ch phm trn cn cha n nh v ng nht. Do , kt qu lachncatinycthcsdngchoccnghincutiptheo nhm bo tn, nhn ging v khai thc c hiu qu ngun dc liu qu trong iu tr T. 38 KT LUN 1.nhgictcdnghglucosemucalccmuthuhi thucbnloiGymnemasylvestre(Retz.)R.Br.exSchult,Gymnema latifoliumWall.exWight,GymnemayunnanenseTsiang,Gymnema inodorum (Lour.)Decne VitNam vi liu 10 g/kgtrnmhnhgytng glucose mu thc nghim bng STZ. Tt c cc mu ca cc loi Gymnema u c tc dng h glucose mu. Trong,loiGymnemasylvestrecbamuPhYn,QungNinh,Thi Nguyn cho tc dng tng ng gliclazid 20 mg/kg. Tng t, G.latifolium cngctcdngttvi4muctcdngtngngvigliclazid20 mg/kg.CcmucahailoicnliG.yunnanensevG.inodorumchotc dng thp hn c ngha so vi gliclazid 20 mg/kg.Ccmuchomchglucosemucaonhtcatngloi:G.sylvestre PhYn(42,24%),G.latifoliumHaBnh(44,24%),G.inodorumHNi (32,82%), G.yunnanense Kon Tum (22,03%).2.La chn c mt s mu i din c tc dng h glucose mu u th trongchiGymnemaR.Br.VitNam:G.sylvestreCT,G.sylvestrePh Yn,G.sylvestreThiNguyn,G.latifoliumThiNguyn,G.latifoliumHa Bnh, G.latifolium Tuyn Quang, G.latifolium Gia Lai. KIN NGH Nghin cu pht trin cc mu c tc dng h glucose mu u th trong chiGymnemaR.Br.lmnguynliusnxutthuchocthcphmchc nng,gpphnaraccsnphmchmscsckhechonhndnvi hiu qu cao v gi thnh hp l. 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