:. /..... 药学与临床研究 年 月 PharmaceuticalandClinicalResearch, ( ) 韩潇媛, 钱之玉 *, 张荣, 陈真, 闫俊岭, 杨丽娜 中国药科大学药理教研室, 南京 : 比较口服西红花酸和西红花苷对异丙肾上腺素 (ISO) 所致原代心肌细胞损伤和大鼠心肌缺血的影响 : 培养原代乳鼠心肌细胞, 用 ISO 诱导心肌细胞损伤, 给予含药血清, 测定 LDH CK 和 MTT; 用 ISO 诱导大鼠心肌缺血模型, 灌胃给药, 测定心电 生化指标和病理改变 : 西红花酸血清减少心肌细胞跳动变化和 CK LDH 释放, 增加细胞活力, 而西红花苷血清对上述指标没有影响 ; 灌胃西红花酸能对抗 ISO 引起 J 点和 T 波抬高, 降低 CK LDH MDA, 升高 SOD, 改善心肌病理变化, 而灌胃西红花苷对上述指标没有作用 : 西红花酸对 ISO 所致心肌细胞损伤和心肌缺血有改善作用, 而西红花苷不具有该作用 提示临床治疗心肌缺血应口服西红花酸, 西红花苷不适合口服 西红花酸 ; 西红花苷 ; 心肌缺血 ; 心肌细胞 ; 血清药理学 ; 异丙肾上腺素 R. ;R + A - ( ) - - Comparisonofefectonisoproterenol-induced myocardialinjurybetweencrocetinandcrocin HANXiao-yuan, QIANZhi-yu *, ZHANGRong, CHENZhen, YANJun-ling, YANGLi-na DepartmentofPharmacology, ChinaPharmaceuticalUniversity, Nanjing, China Abstract Objective:Tocomparetheefectofcrocinandcrocetinonisoproterenol-inducedmyocardialinjuryandischemiainprimeculturedcardiacmyocytesofrats.Methods:Aninvitroinjurymodel ofratneonatalcardiacmyocyteswasinducedbyiso( mm L - ).Celsweretreatedwithserumfrom ratsgivencrocetin(, mg kg - )orcrocin( mg kg - )for h.thelevelofcreatinephosphokinase(ck)andlacticdehydrogenase(ldh)inculturemediumweremeasuredandthecelactivitywasdetectedbymttasay.inaddition, myocardialischemiawasinducedbyiso( mg kg - for twodays, onceaday, sc)invivo.theratswererandomlyigcrocetinorcrocin.subsequently, Jpoint shiftandtwavechangewereasesedintheelectrocardiograph(ecg)afterthetreatment.cardiac indexeswereexamined.thelevelofldh, CK, malondialdehyde(mda)andsuperoxidedismutase (SOD)weremeasured.ThehistopathologicalchangeofmyocardialwasinvestigatedbyHEstaining. Results:Comparedwithmodelgroup, crocetincontainedserumreducedchangeofbeatingfrequency andleakageofldhandck, andprotectedtheactivityofcardiacmyocytes.crocetin(ig)downshiftedthejpointandtwavechange, obviouslydecreasedthelevelofldhandckinplasmaandthe contentofmdainhearttisueandincreasedtheactivityofsod.thehistopathologicalchangesconfirmedtheprotectiveefectofcrocetinonthemyocardialinjury.however, igcrocingavenosignificant improvementofisoproterenol-inducedmyocardialinjury.conclusion:crocetin(ig)canimproveisoproterenol-inducedmyocardialinjuryinvitroandinvivowhileigcrocincannot.itissuggestedoraladministrationofcrocetincanbeusedtocuremyocardialischemiainclinic, whilecrocinoforaladministrationisnotsuitableforthisuse. Keywords Crocetin;Crocin;Myocardialischemia;Cardiacmyocyte;Serumpharmacology;Isoproterenol,, E-mail:hanxiaoyuan @.com *,,,, Tel: - E-mail:qianzhiyu@ com
第 期 韩潇媛等, 比较西红花酸和西红花苷对异丙肾上腺素损伤心肌的作用 (CrocussativusL.) ( -, - - ),,,, HPLC, ;, HPLC, [ ],,. % ±. %,. % ±. % ( ),. % ±. %,. % ~. %,,, [ ],,,, [ ],, ISO, ISO ; ( ),,. 药品与试剂 (ISO, Sigma); - - - (BrDU, Sigma); (, : ); (, : );DMEM (Gibco); (MTT, Amresco); (PPA); (Typsin, Gibco); (LDH) (CK) (MDA) (SOD) ( ), ;,,. %, %(HPLC ),. %CMC-Na. 仪器 (ECG-, ); (Thermo); Eppendorf R. ISO 诱导乳鼠心肌细胞损伤模型.. 乳鼠心肌细胞的分离培养 ~ d SD ( ),,, D-Hanks, mm,, ml. %, min, ml % DMEM, ~,, r min - min,, D-Hanks, % DMEM,,, ml h( ),, r min - min, % DMEM - - - (. mmol L - ), ml -, μl,, h, % DMEM,, d.. 含药血清制备 SD, g~ g, : ( mg kg - ) ( mg kḡ, ),, ig. % CMC-Na h,,, - [, ], HPLC,.. 不同含药血清对 ISO 造成心肌细胞损伤的影响 d,, ( ml % DMEM ), ISO [ ]
药学与临床研究 PharmaceuticalandClinicalResearch 第 卷 (. ml - mol L - ISO +. ml % DMEM ), (. ml +. ml - mol L - ISO +. ml % DMEM ), (. ml - mol L - ISO +. ml % DMEM ), (. ml - mol L - ISO +. ml % DMEM ), [. ml - mol L - ( > %, HPLC )+. ml - mol L - ISO +. ml % DMEM ] [. ml - mol L - ( > %, HPLC )+. ml - mol L - ISO +. ml % DMEM ] h,, LDH CK μlmtt, h,, μldmso, nm (A). ISO 诱导大鼠急性心肌缺血模型 SD,, g~ g, Rona [, ], ISO :,, ( g kg -,, mg kḡ ) ( mg kg -, ) ig, bid,. %CMC-Na sc, sciso ( mg kḡ, qd, d, h ) %, ISO min, T J (QRS T, ST ), ΔT ΔJ,, -,, %, HE, MDA SOD LDH CK MDA SOD,. 统计方法 SPSS., x±s, t. 各组血清血药浓度 HPLC, (. ±. )μg ml -, - mol L -, [ ],, HPLC. A B C, D E A D, B, C E,,, Fig. HPLCchromatogramsofcrocinandcrocetinsera byoraladministratedofcrocinorcrocetin. Blankserum(A), crocetinserum(b)andcrocinserum (C)were detectedforcrocetin.andblankserum(d)andcrocinserum(e)were detectedforcrocin.peak, areimpurities.peak, arecrocetin.. 对 ISO 损伤心肌细胞 MTT 的影响, (P<., P<. ) (P<. ), (P>. ). 对 ISO 损伤心肌细胞生化指标的影响, LDH CK (P<. ), CK (P<. ), LDH LDH CK CK (P<. ), LDH
第 期 韩潇媛等, 比较西红花酸和西红花苷对异丙肾上腺素损伤心肌的作用 LDH CK, (P>. ) ISO LDH CK (x±s, n= ) Table EfectofCrocetinandCrocinonleakageof LDH, CKandMDAcontentandrelativecelactivity incardiacmyocytesinjuredbyiso(x±s, n= ) LDH (U L - ) CK (U ml - ) MTT (%). ±. *. ±. **. ±. **. ±.. ±.. ±.. g L -. ±.. ±. **. ±. * mg kg -. ±.. ±. **. ±. * - - mol L. ±.. ±. **. ±. * mg kg -. ±.. ±.. ±. - - mol L. ±.. ±. **. ±. ** * P<., ** P<. vs ; P<. vs. 对大鼠心电图和心脏指数的影响,, ISO J T (P<. ) (P<., P <. ) T J, ( / ) (P<. ); (P<., P <. ); (P>. ) (P<. ) ISO (x±s, n= ) Table EfectofCrocetinandCrocinonelectrocardiogramand myocardialindexofmyocardialischemiaratsinducedbyiso(x±s, n= ) (mg kg - ) ΔJ (mv) ΔT (mv) (%) ±. ** ±. **. ±. **. ±.. ±.. ±.. ±. *. ±. **. ±. **. ±. *. ±. **. ±. *. ±.. ±. **. ±.. ±.. ±. *. ±. * P<., ** P<. vs ; P<. vs. 对血浆 LDH CK 水平和心脏组织匀浆 MDA SOD 的影响, LDH CK (P<. );, LDH (P<., P<. ), CK. %(P<. ); LDH CK ; LDH CK (P<. ) sciso, MDA (P<. ), SOD. %(P<. ) ISO MDA SOD (P<., P<. ) ISO MDA SOD, MDA, SOD ISO LDH CK MDA SOD (x±s, n= ) Table EfectofCrocetinandCrocinonlevelofLDH, CKinplasmaandMDAcontent, SODactivityinmyocardialhomogenateofmyocardialischemiaratsinducedbyISO(x±s, n= ) (mg kg - ) LDH (U L - ) CK (U ml - ) MDA (nmol mgprot - ) SOD (U mgprot - ). ±. **. ±. **. ±. **. ±. *. ±.. ±.. ±.. ±.. ±. *. ±. *. ±. **. ±. **. ±. **. ±. **. ±. **. ±. *. ±. *. ±. *. ±. *. ±. *. ±.. ±.. ±.. ±. * P<., ** P<. vs ; P<. vs. 病理改变 %, HE,,, ;,, ;,,,
药学与临床研究 PharmaceuticalandClinicalResearch 第 卷. ( ) (A) ;(B) ;(C) ; (D) ;(E) ;(F). Fig. HEstainofasectionofcardiacmuscle. (magnification ):(A)Normalgroup;(B)Modelgroup; (C)FuFangDanShengroup;(D)Crocetingroup( mg kg - ); (E)Crocetingroup( mg kg - );(F)Crocingroup. β,, ISO,, [ ], ISO,,, HPLC, ;,,, ISO [, ], ISO,,,, LDH CK ;,,, LDH CK,, HPLC,, ISO, ;,,,,,,,,, ISO,, ISO J T, LDH CK MDA, SOD, ( ), ISO ;,, ISO,, (ig mg kḡ, h, μg ml - [ ] );,,,, ( ),,,,,,? [ ],,,,,,,, [ ],,.RP-HPLC [ J]. 药物分析杂志,, ( ): -. [ ],. [ J]. 中 国新药杂志, ; ( ): -. [ ],. [ J]. 中草药,, ( ): -. [ ] RonaG, ChapelCI, BalazsT, etal.aninfarct-likemyocardial lesionandothertoxicmanifestationsproducedbyisoproterenolin therat[ J].AMAArchPathol,, ( ): -. [ ],,,. [ J]. 中国药理学通报,, ( ): -. [ ] CommunalC, SinghK, PimentelDR, etal.norepinephrinestimulatesapoptosisinadultratventricularmyocytesbyactivationofthe beta-adrenergicpathway[ J].Circulation,, ( ): -. [ ] Iwai-KanaiE, HasegawaK.Intracelularsignalingpathwaysfor norepinephrine-andendothelin- -mediatedregulationofmyocardial celapoptosis[ J].MolCelBiochem,, ( - ): -. [ ] ShengL, QianZ, ZhengS, etal.mechanism ofhypolipidemic efectofcrocininrats:crocininhibitspancreaticlipase[ J].Eur JPharmacol,, ( - ): -. ( : - - )