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1、h枸杞葉總黃酮對人肝癌細(xì)胞HepG2增殖與凋亡的影響#雷蕾1，袁國強(qiáng)2，劉健鋒1，王長江1，岳明霞1，秦崧1，張璐1，王進(jìn)寶1，邵寶平51，王建林1**（1.蘭州大學(xué)生命科學(xué)學(xué)院，蘭州730000；2.蘭州大學(xué)第二醫(yī)院神經(jīng)病學(xué)研究所，蘭州730030）摘要：【目的】研究枸杞葉總黃酮對體外培養(yǎng)的人肝癌細(xì)胞HepG2增殖抑制及其細(xì)胞凋亡的1015202530354045影響?！痉椒ā渴褂貌煌瑵舛鹊蔫坭饺~總黃酮作用于體外培養(yǎng)的人肝癌細(xì)胞HepG2，用四甲基偶氮噻唑藍(lán)(MTT)法檢測HepG2細(xì)胞存活率,通過Hoechst33258染色法，AV/PI雙染
2、流式細(xì)胞術(shù)檢測HepG2細(xì)胞凋亡情況,利用免疫細(xì)胞化學(xué)法分析枸杞葉總黃酮對HepG2細(xì)胞凋亡相關(guān)蛋白Bax、caspase-3表達(dá)的影響。【結(jié)果】枸杞葉總黃酮分別作用于人肝癌細(xì)胞HepG248h、72h均可明顯抑制其增殖，并呈現(xiàn)顯著的時間、劑量依賴性。高濃度枸杞葉總黃酮作用人肝癌細(xì)胞HepG248h后，HepG2細(xì)胞呈現(xiàn)出典型的細(xì)胞凋亡特征，凋亡率可高達(dá)67.3%。枸杞葉總黃酮作用于HepG2細(xì)胞一定時間后，可引起促凋亡蛋白Bax、caspase-3的表達(dá)上調(diào)?！窘Y(jié)論】枸杞葉總黃酮可明顯抑制人肝癌細(xì)胞HepG2增殖和誘導(dǎo)細(xì)胞凋亡,其作用機(jī)制可能是通過有效
3、上調(diào)促凋亡蛋白Bax，破壞細(xì)胞內(nèi)穩(wěn)態(tài)環(huán)境，繼而激活凋亡蛋白caspase-3，最終引發(fā)人肝癌細(xì)胞HepG2凋亡。關(guān)鍵詞：人肝癌細(xì)胞HepG2；枸杞葉總黃酮；凋亡；Bax；caspase-3中圖分類號：Q95EffectsoftotalflavonoidsinLyciumchinensisleavesonproliferationandapoptosisofhepatomacelllineHepG2invitroLEILei1,YUANGuoqiang2,LIUJianfeng1,WANGChangjiang1,YUEMingxia1,QINSong1,Z
4、HANGLu1,WANGJinbao1,SHAOBaoping1,WANGJianlin1(1.SchoolofLifeScience,LanzhouUniversity,LanZhou730000;2.Instituteofneurologyofthesecondhospital,LanZhouuniversity,LanZhou730030)Abstract:[Aims]ThegoalofthisstudywastoinvestigatetheeffectsoftotalflavonoidsinLyciumchinensisleavesoncellp
5、roliferationandapoptosisofhumanhepatocellularcarcinoma(HepG2cells)culturedinvitro.[Methods]HepG2cellsweretreatedwithdifferentdosesoftotalflavonoidsinLyciumchinensisleaves,thecellviabilitywasmeasuredusingMTTassay.ThemorphologicalchangesoftheapoptoticcellswereassessedbyfluorescentH
6、oechst33258stain.Theapoptoticratewasmeasuredbyflowcytometry.Theapoptosis-associatedproteinexpressionlevelsofBaxandCaspase-3wereanalyzedbyimmunocytochemistrymethod.[Results]ThetotalflavonoidsinLyciumchinensisleavecandose-dependentlyandtime–dependentlyinhibitthecellviabilityofHepG2
7、effectivelywhenincubatedeitherfor48horfor72h.TheHepG2wastreatedwithtotalflavonoidsinLyciumchinensisleavesinhighdosefor48h,thecellpresentedtypicalapoptosischaracteristic.Themaximumratioofapoptosisisupto67.3%.Thepro-apoptoticproteinbaxandcaspase-3wereincreased,whenthecellwasincubat
8、edforacertaintime.[Conclusion]Thetotalfl
