白藜芦醇体外及小鼠体内抗白血病作用机制.doc

白藜芦醇体外及小鼠体内抗白血病作用机制【摘要】目的:探讨白藜芦醇(resveratrol,Res)体内外抗白血病作用的分子机制。方法:采用HuT78、Jurkat和L12103种白血病细胞株,不同浓度Res作用不同时间后,应用甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)比色法检测细胞增殖情况,流式细胞仪检测细胞凋亡,蛋白印迹法检测Bcl2和Bax蛋白表达,免疫沉淀法检测信号转导子和转录激活子(signaltransducersandactivatorsoftranscription,STAT)3及其磷酸化活性(phosphoTyr705STAT3,pSTAT3)。BALB/、25、50mg/(kg·d)Res组,皮下注射L1210细胞建立白血病模型。逆转录聚合酶链式反应(reversetranscriptionpolymerasechainreaction,RTPCR)检测各组白血病小鼠外周血T淋巴细胞白细胞介素6(interleukin6,IL6)基因表达,流式细胞仪检测IL6蛋白分泌,免疫组织化学法和蛋白印迹法检测肝脏活性pSTAT3蛋白表达。结果:Res能抑制白血病细胞HuT78、Jurkat、L1210的生长增殖,诱导细胞凋亡,同时明显下调Bcl2/Bax比值、降低活性pSTAT3的蛋白表达;Res还能够剂量依赖地下调白血病小鼠外周血T淋巴细胞IL6mRNA表达及其胞内分泌,同时降低小鼠肝组织中活性pSTAT3蛋白表达水平。结论:Res体内外均具有明显的抗白血病效应,其作用机制可能与其对IL6及pSTAT3的影响有关。【关键词】白藜芦醇;细胞凋亡;白血病;白细胞介素6;STAT3转录因子;信号转导接头蛋白类;小鼠;体外研究 Methods:Threekindsofleukemiacelllines,HuT78,JurkatandL1210cells,,methylthiazolyltetrazolium(MTT)colorimetrywasusedtodetectthecellproliferation;apoptosiswasdetectedbyflowcytometry;WesternblotandimmunoprecipitationmethodwereusedtodetecttheBcl2andBaxproteinsexpressionandtheactivityofphosphosignaltransducerandactivatoroftranscription3(pSTAT3).Besides,atotalof40BALB/,25and50mg/(kg·d),leukemia6(IL6)hesecretionfunctionofTlymphocytesofthemice;theIL6mRNAexpressioninthelivertissueofmicewasalsodetectedbyreversetranscriptionpolymerasechainreaction(RTPCR),andtheexpressionofthepSTAT3proteinwasmeasuredbyWesternblotandimmunohistochemicalmethod. Results:TheresultsindicatedthatresveratrolcouldinhibittheproliferationofHuT78,,theradioofBcl2/Baxandexpressionofp,resveratrolcouldreducetheexpressionofIL6mRNAandintracellularcontentofIL6,anddecreasetheexpressionofpSTAT3proteinintheliverofleukemicmiceatadosedependen