B YTTRNG I HC Dc H NIV TH THANH HNGNGHIN CU TC DNG CA DCH CHIT L BNG LNG NC(Lagerstroemia speciosa (L.) Pers) TRN NNG ACID URIC MU V HOT XANTHIN OXIDASEGAN CUA CHUT THC NGHIM KHALUNTT NGHIP DC sNgi hng dn:1.TS.Phng Thanh Hng2.DS,Hong Vn Giang Ni thchin:BmnHosinh Trng ihc DcHNiiKLiNtCHANT H l / V f \ l N o y A v . . . t h n g ,,. . ( . V r r i 20. 11, . HNI-2011'^ 'LI CMNVi lng bit n chnthnh v su sc nht,ti xnc gi li trinn TS.PhngThanhHng,DS.HongVnGiang ngi trctiphng dn, tn tnh gip ti thc hin kha lun ny.Ti xnchnthnhcmnnhng kinng gpcaccthycvs gipnhittnhcaccanhchk thutvintiBmnHosinh,Bmn Dclc,trng ihcDcH NivVindcliutrung ng lun hng dn,ngvinvgiptitrong sut qutrnhlmthcnghim.S tntmcng kinthcuynbccaccthyc,lunlunltmgng sng chotinoitheotrong qutrnhhctp,nghincutrong hintivctng lai.Ti xn gilicmnn gianh,bnbti,nhng ngi lunlun gip ,ng vin v to iu kin choti hon thnh kha lun ny.H Ni,thng 5 nm 2011 Sinh vin VTh ThanhHngMCLCLi cm n Mc lcDanhmc cck hiu,ccch vit tt Danhmc ccbng Danhmc cchnhT VN l Phn1;TNG QUAN31.1.TNG QUAN VBNH GT..................................................................... 31.1.1.Lch s bnh g t ...............................................................................................31.1.2.nh ngha.........................................................................................................31.1.3.Dch t ............................................................................................................... 41.1.4.C ch bnh sinh ca g t ................................................................................ 41.2.XANTHIN OXIDASE...................................................................................... 61.2.1.Phn b ..............................................................................................................61.2.2.c im cu trc v c ch hot ng......................................................... 61.2.3.Vai tr ca x otrong chuyn ho acid uric.................................................... 71.3.CC NHM THUC IU TR GT....................................................... 81.3.1.Thuc c ch phn ng vim........................................................................... 81.3.2.Thuc gy tng thi acid uric mu................................................................. 101.3.3.Thuc gy tng oxy ho acid uric mu.......................................................... 111.3.4.Thuc c ch XO............................................................................................. 111.3.5.Thucc ngun gc t dcliu...................................................................121.4.CYBNGLNGNc.................................................131.4.1.c im hnh thi..........................................................................................131.4.2.Phn b ............................................................................................................ 141.4.3.B phn dng............................................................................................... 141.4.4.Thnh phn ha hc.................................................................................... 141.4.5.Cng dng.................................................................................................... 151.4.6.Cc nghin cu v tc dng ca bnh ca Bng lng nc.................... 15Phn 2:I TNG V PHNGPHP NGHINcu............. 172.1.NGUYN VT LIU V THIT B NGHIN cu...................... 172.1.2.Dc liu......................................................................................................172.1.2.Thuc th v ha cht................................................................................ 172.1.3.ng v t ...................................................................................................... 172.1.4.Thit b v dng c ..................................................................................... 172.2. NI DUNG NGHIN c u .......................................................................182.3.PHNG PHP NGHIN CU............................................................ 182.3.1.Phng php chit xut dc liu............................................................. 182.3.2.Phngphpnhginhhngcadchchitnnngaciduricmu trn chut bnh thng.................................................................................. 182.3.3.Phngphpnhginhhngcadchchitnnngaciduricmu trn m hnh gy tng acid uric bng kalioxonat........................................ 192.3.4.Phngphpnhginhhngcadchchitlnhotxanthinoxidase gan chut nht trng bnh thng.......................................................202.3.5.Phngphpnhginhhngcadchchitlnhotxanthinoxidase gan chut nht gy tng acid uricmu bng kali oxonat.................. 202.3.6.Cc phng php nh lng ccch tiu ha sinh..................................202.3.7.Phng php x lsliu..........................................................................23Phn 3:THC NGHIM,KT QU V BN LUN................................243.1.Thc nghim vkt qu............................................................................ 243.1.1.nh hng ca dch chit l BLN trn chut bnh thng........................243.1.2.nh hng ca dch chit l BLN trn chut gy tng acid uric mu bngkali oxonat................................................................................................................ 303.2.Bnlun..........................................................................................................343.2.1.nhhngcadchchitlBnglngncnnngaciduricmu............................................................................................................................343.2.2.nhhngcadchchitlBnglngncnhotenzymxo gan chut thc nghim............................................................................................ 34KT LUN V XUT.................................................................................. 36TI LIU THAM KHOTN CCK HIU, CH VIT TT1.BLN:bng lng nc.2.DHBS:3,5-dichloro-2-hydroxy benzen sulfonic acid.3.T:i tho ng.4.XO;xanthin oxidase.5.COX:cyclo-oxygenase.6.DL:dcliu.7.SE;sais chunDANH MCCC BNGTnbng TrangBng 3.1Anh hng ca dch chit BLNln nng acid uric ca chut nht trng bnh thng26Bng 3.2AnhhngcadchchitBLNlnhotxanthin oxidase gan chut nht trng bnh thng29Bng 3.3AnhhngcadchchitBLNlnnngaciduric ca chut nht trng gy tng acid uric bng kali oxonat31Bng 3.4AnhhngcadchchitBLNlnhotxanthin oxidaseganchutnhttrnggytngaciduricbng kali oxonat33DANH MCCC HNHTnhnh TrangHnh1.1 V tr cc khp b vim,cc ht tophi trong bnh Gout 6Hnh1.2 C ch phn ng ca enzym xanthin oxidase 7Hnh1.3 Cy Bng Lng Nc 15Hnh 3.1Anh hng ca dch chit BLNln nng acid uric ca chut nht trng bnh thng27Hnh 3.2AnhhngcadchchitBLNlnhotxanthin oxidase gan chut nht trng bnh thng30Hnh 3.3nhhngcadchchitBLNlnnngaciduric ca chut nht trng gy tng acid uric bng kalioxonat32Hnh 3.4AnhhngcadchchitBLNlnhotxanthin oxidaseganchutnhttrnggytngaciduricbng kali oxonat34T VN Gt l mt bnh ri lon chuyn ha acid uric vi cc triu chng in hnh l vim,auquanhcckhpxng.Bnhkhngchgyaun,nhhngtikh nnglaong,hn th na nu bnhkhng ccha tr kpthic thdn ti bin chngtnph[12].ccncphttrin,bnhgtchimtlt1 -2%.TiVit Nam, cng vi s pht trin ca cc iu kin kinh t - x hi, bnh ngy cng hay gp. Trong mt nghincu v m hnh bnh tt tiKhoa c xomg khp-Bnh vin Bch Mai trong10 nm (1991- 2000), bnh chim tl8% (so vi trc y l1,5%),ng hng th 4 trong cc bnh khp hay gp ti khoa [8].Mc tiu quan trng trong iu tr gt l lm gim nng acid uric trong mu. cnhiuloithuctrnthgiiraiciutrrngrivemlihiuqu caonhngasl nhngthuctnghpha hc,khidngkodithngcnhiu tcdng khng mongmun.Dovicnghincunhng thuccngungcthin nhin hn ch tcdng khng mong mun khidng kodiangl xu hng pht trincayhcthgiidatrnnntrithcyhcctruyn.Aciduriclmtsn phmchuyn ho ca cc base purin.Ccbasepurin c chuyn thnh hypoxanthin v xanthin ri b oxy ho nh xc tc ca enzym xanthin oxidase (XO) thnh acid uric. Nh vy xo l mt trong nhng enzymquan trng tham gia vo qu trnhsn sinh ra aciduric.Vvy,mttrongnhnghngnghincuiutrgthinnayls dng cc dc liu c tc dng c ch xo lm gim nng acid uric mu [6].Theo y hc c truyn,c nhiu dc liu c s dng iu tr gt. Trong s,Bnglngnc(BLN)(Lagerstroemaspeciosa(L.)Pers.)csdng theo kinh nghim dn gian v c lu hnh rng ri nh mt thc phm chc nng iu trgt nhiu nctrn thgiinh Philippin,ThiLan,Nht Bn...Trong khi Vit Nam, Bng lng nc (BLN) mc d c trng kh ph bin nhng ch mi cbitnvicngdnglmcycnh.TcdngchagtcaBLNVit Nam hunhchac nghincu.Vildo,chng titin hnh ti"NghincutcdngcadchchitlBnglngnc{Lagerstroemiaspeciosa(L.)Pers.)trn nng acid uric mu v hot xanthin oxydase gan ca chut thc nghim" vi cc mc tiu sau:-nh gi tc dng h acid uric mu ca dch chit l Bng lng nc trn chut bnh thng v chut gy tng acid uric mu bng kali oxonat.-TmhiucchgyhaciduricmucadchchitlBLNthngquanh hng trn hot xanthin oxidase gan chut thc nghim.Phn1:TNG QUAN1.1. TNG QUAN V BNH GT1.1.1. Lch s bnh gtGtl mt trong nhng bnhlc bitnlui nht ca loingi.Tn bnh t gcch Latinhlgotta(ngha l ng vn thnh git)haygcch HiLpl podagra (podos: bn chn, agras:ng vn)[1].Miticuithk18,ccnhkhoahccvMmiphthinccc tinh th urat trong cc u cc (tophi)quanh khp, trong cc vin si h tit niu,ng thi pht hin cs khc nhau gia lngacid uric nc tiu ngi bnh thng v ngi bnh.n nhng nm 60ca th k 20,cc nh khoa hc nghin cu su hn vsinh hc t bo,cchsinhl bnh ca tnh trng tng aciduricmuv bnh gt. Nm1679, Anton von Leewenhoek, ngi pht minh ra knh hin vi,l ngi u tin nhn thy tinh th urat, nhng sau vi nm Alfred Gaod mi khng nh"urat l nguyn nhn ca vim gt". Nm1931, Gaod Archibaldtuyn b nguyn nhn ca gt l do rilon s chuyn ha"[28],Do ,ngi ta bit rng gt l bnh l do rilonchuyn ha gy ra bitnh trnglng ngcc tinh thurat cc mcac th do tng acid uric trong mu[1].T cc phng php chn on v iu tr bnh gt c cng c v hon thin.1.1.2. nh nghaBnhgtlmt bnhkhpdorilonchuyn ha nhnpurinca tbo,lm tngcaoqumcacidurictrongmuv trongccmcac th,lngng tinhth urat trong cc t chc nh mng hot dch khp,sn xng, gn, t chc di da.c trngcabnhlnhngtvimthpkhpcp,tiphtgyaud di,cthtin trin thnh vim khp mn tnh[1],[8].Y hc c truyn cn gi gt l thng phong,do ngoi t xm nhp vo c th gy tc nghn kinhlc, kh huyt tr ti khp gy au,co dui kh khn.Khiu,bnhcn c biu kinhlc,bnhlu, t kh vogn xng gy tn thng tng ph,chc nngcakhhuyttndchrilon,tndchtrthnhm,khhuytngngtr thnh , m kt m hnh thnh cc u cc tphi quanh khp, di da. Bnh tin trin lungygytnthngncanthn,lmbindngcckhpvtiphtnhiuln[5],[11].1.1.3. Dch t.Gt ch yu gp nam gii, t l gp cao nht tui trn 40, trc y bnh t khi xy ra ngi tr nhng ngy cng c xu hng tr ha, n t khi xy ra trc tuimn kinh.Tlbnhnhn namchimkhong90%cc trng hp.Bnhchim khong0,1-0,2%tngsbnhnichungv0,4-5%tngsbnhkhpv thngc lin quan n mc sng v ch dinh dng hng ngy[5].nhng nccnn kinht pht trin,bnhgt chim tlcaovokhong1- 2%.TiVit Nam,cng vis phttrinca kinh t- x hi,khong10nmtrli ybnh c chiu hng tng ln r rt.Da trn mt c s d liu bo him ca M c tnhrng t nm1990-1999,tlbnhgt tng60% nhng ngi trn65tui v tng gpivi nhng ngi trn75tui.Ccnghin cu khccho thy bnh gt ngycngtr nnphobintrongccx hinh NewZealand,iLoanvHoaK [41].TheomtnghincukhccaAnh,tlmcbnhgttrongsngitrng thnhl1,4%,cbittptrung namgii trn75tui.Nhngconsnychorng bnh gtl dng bnh ph binnht ca vim khp ngiln v ang tip tc tng ln.XuhngnykhngchcquanstthyphngTymcnccnc ang pht trin Chu [31].1.1.4. C ch bnh sinh ca gt1.1.4.1.C ch lng ng acid uric.C chlngngaciduricch yuldotngaciduricmukodidnns bin i v hnh thi cc t chc. Tng acid uric dn n kt ta thnh cc tinh th hnh kimgytnthngsn,mnghotdch,baokhp,hnhthnhcchttophigy phhyxngdidngkhuytxnghnhcu.Cctnthngthphtnh:vim mnghotdch,tngsinhmnghotdch,thmnhimcctbolympho.Slng ng cc tinh th tchc to thnh cc ht tophi kch thc to nh khc nhau.Lng ngtinh thurat kthndnn tnthng thnnh:sithn,vimthnk,x ha cu thn,lng acid uric trong nc tiu tng v s toan ha nc tiu dn n si titniutrongbnhgt.Vkhitn thnglanrngdnnsuythn,tnghuytp [8],1.1.4.2.C ch bnh sinhcn gt cp tnhTngaciduricmudnntngnngvkttacctinhthacidurichoc mui ca n trong tchc v dch c th.Gii hn ha tan ti a ca acid uric trong mu khng qu 70 mg/L (tng oTig khong 415|xmol/L)[1].Khi vt qu nng ny acid uric d b kt ta di dng tinhth hnh kim cc t chc. Kh nng kt ta caccaciduricmuphthucnhiuyut,trongvicgnviproteinhuyt tngctcdng hnch kt ta.Trongiu kin nhit thp,dchcpHacid th natri urat dkt tinh thnh tinh th,ngcli trongiu kin nhitcaohn v pH ca dch kim th natriurat c ha tan cao hn,t b kt tinh hom.S kt tinh thnh cc tinh th natri urat khi t n mt lTig nht nh s gy ra vim khp (vim khp vi tinh th) v gy ra cn gt cp tnh [5].1.1.4.3.C ch bnh sinhcn gt mntnhGtmn tnhbiuhinbngduhinnicccc(tophi)v vima khpmn tnh.Khi cc ht tisn khp v s khi pht qu trnh vim gy cn gt cp.S lng ng vi tinh th cnh khp, trong mng hotdch, trong m sn v m xng s dn n bnh xng khp mn tnh do gt [8].f Gout Attack Locations,- - ' . . . i f .vrhrj.Hnh1.1. V tr cc khp b vim, cc ht tophi trong bnh Gout1.2. XANTHIN OXIDASEXanthin oxidase (Xanthin oxygen oxidoreductase, EC1.17.3.2),l en2ymxc tc qu trnh oxy ha xanthine v hypoxanthine c th tip tc xc tc cho qu trnh oxy ha xanthin thnh acid uric .1.2.1. Phn b ngi, XO thng c tm thy trong gan v trong mu. cc ng vt khc nhauncphnbrng ritrong muv trongccm.xo cngcphthin trong mt s m ca ng vt c v, chim,cn trng v vikhun[19].ng vt, XOc tm thy trong tbo gan, tbo rut,ngoira cn thy trong t bo thn, ddy,huyt thanhvmphi[4],[21],nng thp thy xng,c,timvno [21].1.2.2.c im cu trc v c ch hot ngXanthin oxidasel mt protein bao gm 2 tiu oTi v ging nhau, c khi lng phnt300kD,mitiunvckhilng150kD.Trongmitiunvc2 phntflavin(FAD),2nguynt molybdenv8nguyn t st gntrnmtnv enzym [35].Qutrnhoxiha xanthinthnhaciduricdinratitrungtmmolybdenca XO[13].Trong trng thi oxi ha, trung tm molybden ca enzym c xy dng nh mhnhLMo'^^OS(OH).Cchcaphnngcchranhsau:protontnhm Mo-OH tn cng trungtminhn tivtrC-8ca c cht xanthin(R),ng thi vnchuynhydrogiiphngratvtrC-8tinhmMo=stora LMo'^0(SH)(OR),OR kt hpvi nguyn t kimloivonhm hydroxyl,sn phm trung gian ny b ph v bis vn chuynelectron ti cc v tr kh khc ca enzym v gii phng H^, to ra LMo^ OS (OR),c pht hin bi quang phcng hng t, sau nhm R c thay th bi hydro, quay v trng thi ban u LMo'^'OS(OH) ca enzym[16].oooHO',p,e'oS_M^S^S-Mo-SH^ ,S-Mo^SsMc=s>=0 >~N >-Ngiu,26i'^0o^Hnh1.2. C ch phn ng ca enzym xanthin oxidase L23.Vai tr ca xo trong chuynho acid uricXanthin oxidase (XO)l enzym ng vai trquan trng trong d ha purin mt s loi, bao gm c con ngi[4], Acid uric l sn phm thoi ha ca cc acid nucleic cbaselpurin.C3nguncungcptinchtcaaciduricgm:thoihaacid nucleic t thc n a vo, t cc t bo bcht v do purin ni sinh trong c th.Qu trnhthoihaacidnucleictoraccpurinnginnhadenin,hypoxathin, xanthin,guanin.Ccpurintrncchuynhathnhaciduricnhcsthamgia caccenzymaminotransferase,hypoxanthinguaninphosphoribosyltransferase,v xanthin oxidase[1],[5].1.3.CC NHM THUC IU TR GT1.3.1.Thuc c ch phn ng vim ColchicinColchicinchitxuttcyColchinumautumnal.Nm1763chphmt Colchicum autummaleln u tin c s dng trong iu tr cn gt cp v n nm 1814 ngi ta mi chit xut c colchicin t loi cy ny[1].Tc dng:Thucchc tcdnggimauvchng vimcpdogt,khngc tcdngvitrnghpkhngphidobnhgt.Tcdngchiuvicongtcp[3].Cch\Colchicinlmgimsdichuyncabchcu,gimstptrungca bch cu ht t chc vim, c ch hin tng thc bo cc tinh th urat v do km hmsnxutlactic,gichopHtichcbnhthcmg.Thuckhngctcdng ln s o thi acid uric. Ngoi ra cochicin cn c tc dng chng phn bo, phn hy lympho bo, tng sc bn mao mch[3].Ch nh: iu tr gt cp, phng cn cp bnh nhn gt mn tnh[3].Tc dng khng mong mun:Thnggprilon tiu ha;bunnn,nn,tiu chy. Ngoi ra thuc gy gim bch cu, tiu cu, ri lon thn kinh c [33]. GlucocorticoidTcdng:Thucctcdngchngvimmnh.Corticoidcnctcdngtrn chuynha(glucid,lipid,protid,chuynhamuinc).Docchcch phospholipasec nnthucgimgiiphnghistaminvccchthahctrunggian gy d ng. c bit, glucocorticoid c ch min dch do gim s lng t bo lympho, c ch chc nng i thc bo v c ch sn xut khng th[3],C ch chng vim:Glucocorticoid c ch phospholipase A2 thng qua kch thch tng hplipocortin,lm gim tng hp leucotrien v prostaglandin. Ngoi ra n cn c tcdng cchdng bchcun nhn,a nhnlymphoboivom gykhi pht phn ng vim [3.Ch nh:iu trgt chng chnhvi NSAIDsv colchicin.Thuc c th iu tr gt cp[33].Tcdngkhngmongmun:Corticoidthnggylongxng,tngng huyt,lotddyttrng,ph,tnghuytp,rionphnbmvccrilon khc[3]. NSAIDsTc dng[3]:-Thucctcdnghst,gimau,chngvim.Lthucgimau,nhng NSAIDsl thuc gim au ngoi vi v khng c tc dng gy nghin.Tc dng chnh trong iu tr gt l tc dng chng vim.- NSAIDs c ch tng hp thromboxan nn c tc dng chng kt tp tiu cu-liu iu tr,thucc tc dng h st khic th bst do bt k nguyn nhn nogynnvchctcdngiutrtriuchng,khnggyh thnnhit ngi bnh thng- Thuc c tc dng gim au t au nh n au va.c bit, tt cho cc chng au do vimC ch:-Thucctcdngcchsinh tng hp prostaglandindocchenzymcyclo- oxygenase (COX) lm gim tng hp prostaglandin.-Thuccnlmbnvngmnglysosomdohnchgiiphngccenzym ca lysosom trong qu trnh thc bo, nn c tc dng chng vim.-Ngoira thuccncchccchttrunggianhahccaqutrnlivim,c chcchtcaenzym,cchsdichuyncabchcu,cchphnngkhng nguyn - khng th.-Ringnhmsalicylatcnlmtnggiiphngsteroidnnlmtngtcdng chng vim[3],Ch nh:iu tr cn gt cp tnh [11],[33].10Tc dng khng mong mun:tcdng khng mongmun ca NSAIDsch yu l:lot d dy t trng, xut huyt,vim thn k[3],[33].1.3.2. Thuc gy tng thi acid uric ProbenecidTc dng:liu iu tr, probenecid c tc dng lm tng thi acid uric (ngc li,liuthplmgimthiaciduric).Ngoira,thuccchvnchuyncc monoamin n dch no ty v c ch bi tit mt [3],[23].C ch:c ch cnh tranh h vn chuyn anion gy c ch ti hp thu acid uric ng thn,lmtngthiaciduricqua nc tiu.Khinngaciduricmugim,cc tinh th urat lng ng cc khp s tan ra v tr li mu ri thi tr dn ra khi c th[3].Ch nh:iutrbnhgtthmnctnthngm,thngphihpvi colchicin v cc thuc chng vim[3].Tcdngkhngmongmun:Probenecidtngianton,tgytcdng khng mong mun.Thng gp ri lon tiu ha, tng to si thn hoc cn auqun thn, d ng v gy bun ng [11]. SuinpyragonSulfinpyrazoncphthinkhongnm1960,trongkhitmkimmtthuc bititaciduricniuvchngvimtcnht.Lchtchuynhaca phenylbutazon.Tcdngvcch'.Tcdnglnsthitrurattngtpronenecid,nhng hiu qu cao hon.Tc dng ca thuc ph thuc vo liu dng.Liu thp lm gim bi xutaciduricvolngngthn,ngcliliucaongnchntihpthuaciduric [23].Ch nh'.Thucdngiutrgtmn,phihpvicolchicinvccthuc chng vim khc[3].11Tc dng khng mong mun:C th gpau bng,bun nn, nn,lot d dy t trng hoc taibin trn mu ging nh phenylbutazon.Dngthucthn trng bnh nhn suy thn[3],[23].1.3.3. Thuc gy tng oxy ha acid uric RasburcaseRasburicaseldngtithpcauratoxidasevcsnxuttchng Saccharomyces cervsiae.Tc dng v c ch: Thuc c tc dng h nng urat. Rasburicase xc tc phn ngoxihacaaciduricthnhchtchuynhaalantoinkhnghottnh,snphm chuyn ha ha tan nhiu trong nc hn acid uric[29].Ch nh:Thuc c dng iu tr gt c sn cng nghim trng [29].Tc dng khng mong mun:Thuc c tnh khng nguyn nn c th gy ra phn ng min dch khi dng thuc. Phn ng min dch c th xy ra nh co tht ph qun, myay,ccphnngdngda,scphnv.Ngoira,thuccngymtstc dng bt li: bun nn, au u,a chy[23],[33]. Pegylat uricase khc phc cc nhc im ca rasburicase, pegylat uricase c pht minh. Tc dng vc ch:Thucc tcdng tt trongh thpnngaciduricmu v gn mc ban u do xc tc qu trnh oxi ha acid uric chuyn thnh alantoin.Ch nh:Thucdng cho bnh nhn gt mn tnh,tht bi vicc phng php iu tr hoc c sn to do tch lng [24].1.3.4. Thuc c ch Xanthn oxdase. AUopurnoTc dng:Thuclmgimnng acid much yudoc chcnh tranhtng hpaciduric.Ngoira,thuclmtngbixutcctinchtcaaciduricquanc tiu[3],[33],12C ch:Allopurinolcchenzymxanthinoxidase,lenzymcvaitrchuyn cctinchthypoxanthinvxanthinthnhaciduric,nhlmgimnngacid uric mu [3].Ch nh: iu tr gt mn, Cc trng hp tng acid uric th pht (do dng thuc chng ung th, thuc li tiu thiazid...)[3].Tc dng khng mong mun:C th gy kch ng tiu ha,c vi gan, suy thn v d ng da [33]. PebuxostatPebuxostatlthucmicavothtrngnhmtthucthayth allopurinol[34],Tc dng v c ch:Thucc tcdng h nng acid uric mu theoc ch c ch xanthin oxidase, tng t allopurinol[34].Ch nh:iu tr gt xut hin vng da , tn thng.c bitl d phng khi khng c colchicin[11].Tc dng khng mong mun:Tc dng khng mong mun thng gp nht khi s dngfebuxostatllmsuygimchcnnggan,tiuchy,auu,bunnn,pht ban, cn gout cp[11].1.3.5. Thuc c ngun gc t dc liuTrong nhng nm gn y, bnh gt ngycng ph bin, nhu cu v thuc cha bnhgtngycngcao.Thuctndciutrgtgithnhcaovgpnhiutc dng khng mong mun.V vy vic nghin cu tc dng iu tr gt c ngun gc t tho dc l cn thit. Vic s dng cc thuc c ngun gc thin nhin c nhiu thun li.ccncnh TrungQuc,Australia,Chile,Paraguay,Panama,Vit Nam,mt snghincuchothynhiudcliucchngminhlctcdngcchhot tnh XO.Mt nghin cu Trung Quc tin hnh trn122cy thuc cs dng trong y hc c truyn iu tr gt, vi liu100)Lig/ml c 69 dch chit methanol v 40 dch13chitncctcdngcchhotxo,trongs29dchchitmethanolv13 dchchitnclmgimhn50%hotxo.Ccchtchottnhcaonhtlm gimhn50%hotx o lQuchi(Cinnamomumcassia),tiplCchoa (Chysanthemumindicnm).lcaLvcopuseiiropoeusvctkhc(Polygonum cuspidatum)[6],[22],NghincugnycaNguynThThanhMaitinhnhvi96cythuc csdngtrongyhcctruyniutrgttiVit Nam.Trongs288dch chit t 96 cy thuc trn th188 dch chit c chng minhc tc dng c ch hot XO mcliu100|Jg/ml,trong 46dchchitlmgim hn50%hotxo, cc dch chit methanol in hnh c tc dng lm gim hn 50% hot x o l: Ngi cu(Artemisia vulgaris),Tmc(Caesalpnia sappan),ibi(Blumea balsamifera), Cchoa(Chrysanthemum sinense),T gic leo(Tetracera scandens)[26].Cc nahin cu nycho thy,nhiu cy thuccs dng iu trgt cda trnCC7ch c ch hot xanthin oxidase.Ngoi racnnhiudcliu khcc tcdng iu trgt docch x o nh: Hy thim{Siegesbeckia orientaisL.)[6],Trcbchdip(Biotaorientalis)[18],[36], Daphne genkwa [27], Du tm {Morus alba L.)[6], Hnh ty {Allium cepa L.)[20].1.4. CY BNG LNG NcBnglngnc(BLN)-Lagerstroemiaspeciosa(L.)Pers.,thuchTvi- Lythraceae.Cy cn c tn khc l T vi tu, tn thng gi l Bng lng [7].1.4.1. c im hnh thiCy gln c kch thc trung bnh.L bu dc, trn gc, nhn ngn chp, di10-20cm, rng 5-9cm,dai, rt nhn,c 2mt l u c mu nht.Chu hoa ng ngn, nhnh c lng, n trn o . Hoa to, rng 3cm hay hn, mu tm; i c lng st;6cnhhoaccung5mm;nhnhiu.Qunangtrndidngtrng(20x18mm) mang l i xo ra, n lm 6 mnh. Ht c ng knh12-15mm[5].14Hnh1.3. Cy Bng Lng Nc (Lagerstroemia speciosa (L.) Pers.)1.4.2. Phn bBLN thng mc vng nhit i, ch yu Nam v ng Nam nh n ,Indonesia,Philipin,Malaysia...[9],ngoira cycnphnb mtsnc Nam M [7].Vit Nam, BLN c trng tng inhiu Ty Nguyn, Bnh Phc, Ty Ninh, ng Nai v mt s thnh ph ln khc nh H Ni, Hi Phng, Nng...Cy mc hoang hoc c trng lm cnh [9].1.4.3. B phn dngBphndngchyucacylv,lvqu{Cortex,FoliumetFructus Lagerstroemiae) [9].1.4.4. Thnh phn ha hcTrn th gii c nhiu nghin cu cho thy thnh phn ha hcch yu ca Bnglngncltanin,chimhn10%,tpchungchyulgivquchn. Trongngchnhtlnhmellagitarmin(gm3chtxcnhcutrcl FlosinB,Lagerstroemin,regininA)vgallotannin(gmacidtannicvpenta-o-15galloyl-D-glucopyranose).Mtdnchtpolyphenolkhccngctmthytrong dchchitacetoncalBLNlvaloneaicaciddilacton[39].MtnghincuVit NamchothyhmlngpolyphenoltonphntronglBnglngncchimti 39,5% tnh theo dc liu kh[7].MtnhmthhaictmthytronglBnglngnclcchpcht terpenoid.TdchchitmethanolcalBnglngncngitaphnlpc mtloitriterpenoidctcdnghglucosehuytlacidcorosolic(acid2a- hydroxyursolic).NgoiralBnglngnccncccterpenoidkhcnh:acid 3p,23-dihydroxy-loxo-olean-12-en-28-oic;tinotufolinc,tinotufolinD,lutein,24- methylencycloartanol acetat, largerenol acetat[7].1.4.5. Cng dngBLNcnhiucngdngcngdngrngrictrongvngoincnh:-V cy c tc dng gim st. Nc sc v cha au bng, tiu chy [2]-R c tc dng sn se, gim st[2]-Ht c cht gy ng[2]-L c tc dng lm gim ng huyt, cha bnh gt [9]-Qu dng p ngoi tr l ming [7].1.4.6.Cc nghin cu v tc dng cha bnh ca Bng lng nc t lu, mt s nc Chu , BLN c s dng nh mt cy thuc dn gian iu tr cc bnh ri lon chuyn ho nh bnh i tho ng, gt.Nm1996,mtnghincu NhtBnchothydchchitncnngcal BLN c tc dng h glucose huyt ng k trn chut nht T typ 2di truyn chng KK-Ay sau 5 tun iu tr.Cng trn chng chut KK-Ay, acid corosolic, mt triterpen t l BLN vi liu 2 mg/kg lm gim nng glucose v insulin huyt tng sau 4 gi ungthuc.Mtnghincu khc tinhnh tiHnQucchothydchchitl BLN16tiuchunhacha1%acidcorosoliclmgim26%glucosehuytcachutT typ 2 chng C57BLKsJ sau 4 tun dng thuc [7].Mtnghincu NhtBnnm2004chothyhotchtacidvaloneaicacid dilacton (VAD)trongl BLNc kh nng c chmnh xanthinoxidase,l enzymc vai tr quan trng trong qu trnh tng hp acid uric ca c th[32]. Nghin cu ny l c s khoa hc bc u cho vic s dng BLN iu tr gt nhiu nc theo kinh nghim ca y hc c truyn.Nh vy, tc dng cha bnh ca l BLN c nghin cu v p dng rng ri tinhiunctrn thgii.Trong khi, Vit Nam,BLNmcdc trng ph bin nhng hu nh cha c bit n trong cha bnh. Nghin cu ca Phng Thanh Hong(2009)chothylBLNctcdng hng huyt trnnhiumhnhthc nghim khc nhau[7],Tuy nhin, cha c mt nghin cu no Vit Nam tm hiu v tc dng iu trbnh gt ca BLN.Do vy, rt cnc nhng nghin cu tip theo nh gi an ton v tc dng cha bnh ca BLN,nhm m ra trin vng khai thc mt loi thc vt ph bin nc ta trong phng v iu tr bnh,c bit l cc bnh ri lon chuyn ha nh i tho Tig v bnh gt.17Phn 2:I TNG V PHNG PHP NGHIN c u2.1.NGUYN VT LIU V THIT B NGHIN c u2.1.2.DcliuLBnglngnc{Lagerstroemiaspeciosa(L.)Pers.)cthuhivo thng5-7nm2010tiH Ni.Saukhithu hidcliuclmsch,phi, sykh,xaythnhbtth,chitbngphngphpngmlnhviethanol70 trong72gi.Mul Bnglngnccgimnhng tn khoa hcbi TS.Trn Th Bch (Vin Sinh thi v Tinguyn sinh vt).2.1.2.Thuc th v ha cht- Kali oxonat (Sigma-Aldrich).-AllopurinolttiuchunBP2001/SP25(YixingCityXingyuMedcine Chemicals Co.,Ltd).-C cht xanthin (Sigma-Aldrich).- B ha cht nh lng acid uric (Teco diagnostics).2.1.3.ng vt-ChutnhttrngthunchnggingSwiss(Mumusculus),chaiging, trng lng 20 2g,khe mnh do Vin V sinh dch t trung ng cung cp- ng vt c nuidng theoch tiu chun ti phng nuing vt th nghim,Trng i hc Dc H Ni.2.1.4.Thit b v dng c- My o quang ph V-VIS- My li tm K centrifuge (Harmonie sris).- My xt nghim sinh ha mu (Teco diagnostics)- T sy chn khng Heraeus-instruments vacutherm-Kelvitront. KUNCI!Dc HN T H U V NM']3y ... .'.'.'. '.h-Vi'j ...i. 'I----^SoOKCB;....18- Ni cch thySSY-H,GFL,Memmert.- Cn k thut Precisa-BJ610C,TE 412 hng Sartorius.-Cn phn tch AY 220 hngShimadzu.- T sy dng c Binder- My nghin ng th v cc micropipet (Labopette)-Ccdngcthnghimphcvcholymuv xtnghim;micropipet, bm kim tim cc c,maoqun,ng nghim ccloi...- Ccdng c dng trong chit xut;bnh chit,ng ong,cc c m...2.2.NI DUNG NGHIN c utcmctiunghincutra,ticthchinviccni dungc thsau:-Chit xut bt dc liu.-nhgitcdnghaciduricmudchchitlBLNtrnchutbnh thng v chut gy tng acid uric mu cp bng kalioxonat.- nh gi c ch gy h acid uric mu ca dch chitl BLN thng qua nh hng trn hot xanthin oxidase gan chut thc nghim.2.3.PHNG PHPNGHIN CU2.3.1.Phng php chit xut dcliuNgmbtdcliuvidungmiethanol70nhitphngtrong72 gi.Dchchitcctthuhidungmidipsutgimthucn.cn c pha thnh hn dch trong nc vi t l1:4 th trn chut[12].2.3.2.Phngphpnhginhhngcadchchitnnngacid uricmutrnchutbnhthigChut nht trngc nuinnh mi trng phng th nghim v c chia ngu nhin thnh ccl,mil10con.19- L chng:ung nc ct.- L ichiu:ung allopurinolliu 20 mg/kg chut nht trng.-Cclth:ungdchchitBLN.3lvi3liukhcnhau:5gDL/kg; 1OgDL/kg,20g DL/kg (kho stliu da trn liu dc liu thng thng)Chutcung mu th trong5ngy vomt gi nhtnh.n ngy thnm,saukhiungthucc2gi,lymu,mulngtnhintrong khongIginhitphngriemlytmvitc3500-4000vng/pht, trong10 pht, thu huyt thanh i nh lng acid uric[11],[14].2.3.3.Phngphpnhginhhngcadchchitnnngacid uricmu trnmhnhgytng aciduricbngkal oxonatSau thnghim nghin cu nh hng ca thucn nng acid uric mu trnchut bnh thng,sla chn liu dng thch hpca dchchitgy h acid uric trn m hnh tng acid uric mu thc nghim[6], B tr th nghimChut nht trng c nuin nh mi trng phng th nghim v c chia ngu nhin thnh ccl,mil10 con.- L chng:ung nc ct.- L i chiu :ung allopurinol viliu 20mg/kg chut nht trng.- L th;ung dch chitl BLN liu 5g DL/kg chut.Chutcungmuthtngngtrong5ngyvomtginhtnh. nngy th nm,timmngbngkalioxonat500mg/kgIgitrclnung thuccuicnggytngaciduricmu.Saukhiungthucc2gi,ly mu,emulngtnhintrongkhongIginhitphngriemlytm vitc 3500-4000vng/pht,trong10 pht,thu huyt thanh inhlng acid uric[11],[14],202.3.4.Phngphpnhginhhngcadchchitlnhotxanthn oxidase gan chut nht trng bnh thngChut nht trng c nuin nh mi trng phng th nghim v c chia ngu nhin thnh cc l,mil10con.- L chng:ung nc ct.- L i chiu:ung allopurinol liu 20tng/kg.-Cclth:ungdchchitBLN.(3lvi3liukhcnhau:5gDL/kg; lOgDL/kg, 20g DL/kg)Cclchutcungmuthtongngtrongnmngyvorntgi nht nh.n ngy th nm,sau khi ung thuc 2gi,git chut ly gan xc nh hot xanthin oxidase[6].2.3.5.Phngphpnhginhhngcadchchitlnhotxanthin oxdase gan chut gy tng aciduricmubngkal oxonatBtrthnghimvtinhnhnhmiutphn2.3.4.Vongyth5, timmngbng kalioxonat500mg/kgIgitrclnung thuccuicng gy tng acid uric.Sau khi ung thuc c 2 gi,chut c mly gan xc nh hot x o [6].2.3.6.Ccphng php nhlng ccch tiuha sinh.2.3.6.1.nhlng acid uric mu.nh lng acid uric mu trong huyt tng bng phng php enzymuricaseAcid uric +O2 +2 H2O -------- Allantoin + CO2 + H2O2PeroxidasH2O2 + 4-Aminoantipyrine + DfBS-----------Chromogen + H2O21Aciduricbchuynthnhallantoinvhydroperoxidnhuricase. Hydroperoxid phnng vi4-aminoantipyrin v 3,5-dichloro-2-hydroxy benzen sulfonicacid(DHBS)tothnhchromogencnhlngbiphngphp o quang bcsng 520nm.-Tlgimnngaciduricmutronghuyttngcalthsovil chng:I(%)=[(Cc -Ct)/Cc]x100%I(%):t l gim nng acid uric huyt thanh ca l th so vil chng Ccinng (mg/dl) ca l chng, c t:nng (mg/dl) ca l th.2.3.6.2.Phng php nh lng x oPhng phpoquang datrnnhlng acd uric:Phng phpcaNoro(1983)[27]Nguyn tc phn ng:Xanthin + O2 + H2O>Acid uric + H2O2 Hnhpphnngbaogmenzymx o vcchtxanthintrongmi trngmphosphatpH8,0ctrong30phtnhit37c.Phnng cngng bngHClIN,bt hotenzym.Hot x o c xcnh thng qua lng acid uric to thnh trong1 pht c o bc sng 290 nm.nvhot(U)caenzymctnhbnglngaciduricsinhra trong1 phttrnIgproteiniukin37c,pH8,0.Hmlngproteintrong gan c nhlng bng phng php Lowry.Cng thc tnh hot th tch:22( Eth"Echng)^ ^>0X1 5/ ml =-------------------------------12,2x0,1Xl , OxTCng thc tnh hot ring:yr/,.u I ml/g protein =Trong :12,2: hp th milimol phn t ca a.uric ti bc sng 290nm (mM''.cm'')6.0:Th tch phn ng (ml)0,1:Th tch enzym cho vo (ml)15:Hs pha long1.0: dy cuvet (cm)T:Thi gian phn ng (60 pht)K:S g protein/lg gan.Xc nh hot tnh x obng phng php chit enzymCnkhongIggan,ctthnhccmnhnh,sauchovongnghin ngth,thmImldungdchmphosphatlnh(pH=8,0)vo.Nghinng thgantrongiukinlnh( t 0,05 4,78%4 L dch chit (1 OgDL/kg chut)325,5513,17 p > 0,05 -7%5 L dch chit (20gDL/kg chut)310,29 33,67 p > 0,05 -2%26oE5.33pfljo-ocz350300250200150100500400'304.06325.55310.29T 289.52217.71Chng AllopurinolDich chitDch chitDch chit5gDL/kglOgDL/kg20gDL/kgLoHnh 3.. nh hng ca dch chit BLNinnng acid urc ca chut nht trng bnh thngNhn xt:- Allopurinol(liu 20mg/kg) c tc dng h acid uric muc ngha thng k so vil chng (p 0,05).273.1.1.2.KtqunhginhhngcadchchitlBLNccnng lnhot xanthin oxdase gan chut nht trng.- B tr th nghim:+ L chng:ung nc ct (n =10)+ L i chiu:ung allopurinolliu 20 mg/kg (n =10)+Cclth:ungdchchitBLN(3lvi3liukhcnhau:5gDL/kg; lOgDL/kg, 20g DL/kg) (n - 10)Chutcungmuth tngngvimiltrong5ngyvomtgi nht nh.nngyth5tinhnhnhlngvsosnhhotenzymxo gan gia ccl.Kt qu c th hin bng 3.2 v hnh 3.2.28Bng 3.2. nhhng ca dch chit BLNlnhot xanthnoxdase ganchut nht trng bnh thngSTT L chutHot enzym xo (mMacid uric/ pht/ g protein gan)% Hot enzymMc c ch1 L chng2,110 0,205100%2 L allopurinol1,2880,233 p < 0,05* 61,01% 38,99%3L dch chit (5gDL/kg chut)1,244 0,078 p < 0,05* p>0,05**58,96% 41,04%4L dch chit (lOgDL/kg chut)1,257 0,075 p < 0,05* p > 0,05** p> 0,05***59,57% 40,43%5L dch chit (20gDL/kg chut)0,934 0,103 p0,05** p>0,05***55,73% 44,26%(*):so snh vi l1;so snh vi l 2;so snh vi l3(Kt qu cho trn bngl gi tr trung bnh ca10chut,migan c tinhnh lpli trong 3ln)292.5T3ns3I I 15S o1o ^ 0l ll | 0.5 qjao- o02.110.934Chng AllopurinolDich chitDich chitDch chit 5gDL/kg lOgDL/kg20gDL/kgLHlnh 3.2. nhhng ca dch chit BLNlnhot xanthn oxidase ganchut nht trng bnh thngNhn xt:-L ung allopurinolliu 20mg/kg chot xo gim c ngha thng k so vi nhm chng (p < 0,05).-CclungdchchitlBLNviccliukhcnhau(5g/kg, lOg/kg,20g/kg) c hot enzym x ogim c ngha thng kso vil chng (p < 0,05),khng khc bit vil ung allopurinol (p > 0,05) v khng khc bit vi nhau (p > 0,05).303.1.2.nhhngcadchchitlBLNtrnmhnhchutgytngacd urcmubng kalioxonat3.I.2.I.nhhngcadchchitlBLNnnngaciduricmutrn mhnhchut tng aciduricmubng kalioxonat.-B tr th nghim:+ L chng:ung nc ct (n =10)+ Lichiu:ung allopurinol 20mg/kg chut (n =10)+ L th:ung dch chit l BLN liu 5g dc liu/kg chut (n =10)Chutcungmuth toTigngvimiltrong5ngyvomtgi nht nh.en ngy th 5tin hnh nh lng v sosnh nng acid uric mu gia cc l.Kt qu c th hin bng 3.3v hnh 3.3.Bng 3.3. nhhng ca dchchit BLNlnnng acd uriccachut nht trng gy tng aciduricbng kali oxonatSTT LNng aciduric mu (.mol/L)p so vi chngT l gim so vi l chng (%)1 L chng 346,25 3,932 L allopurinol242,3115,09 p< 0,01* 30,02%3 L dch chit (5gDL/kg chut)287,28 19,82p< 0,01* p>0,05**17,03%(*):so snh vi l1;(**): so snh vi l 231400350o 3003'TJ250uvl 200p150O-100bOc,050Z0346.25T287.28ChngAllopurinolDch chit5gDL/kgLHnh 3.3. nh hng ca dch chit BLNlnnng acid uric ca chut nht trng gy tng acd uric bng kai oxonatNhn xt:-Lchutungallopurinolcnngaciduricgimcnghathng k so vil chng (p < 0,01)- L ung dchchitl Bng lng nc nng acid uric gimc ngha thngksovilchng(p 0,05)323.I.2.2.nhhigcadchchitlnhotxanthnoxdaseganchut gy tng acduric bngkali oxonat- B tr th nghim:+ L chng:ung nc ct (n =10)+ L ichiu:ung allopurinol 20mg/kg chut (n =10)+ L th:ung dch chit l BLN liu 5g dc liu/kg chut(n =10)Chutcungmu th tngngvimiltrong5ngyvomtgi nht nh.nngyth5tinhnhnhlngvsosnhhotenzymxo gan gia ccl.Kt qu c th hin bng 3.4 v hnh 3.4.Bng 3.4.nhhng ca dch chit BLNlnhot xanthnoxdase gan chut nht trng gy tng acid uric bngkali oxonatSTT L chutHot enzym x o(mM acid uric/ pht/ g protein gan)% Hot enzymMc c ch1 L chng1,250,15100%2L allopurinol0,74 0,04 p