發(fā)布時(shí)間：2018-03-04 05:06

  本文選題：氧化應(yīng)激　切入點(diǎn)：細(xì)胞凋亡　出處：《細(xì)胞與分子免疫學(xué)雜志》2015年01期 　論文類型：期刊論文

【摘要】：目的探討糖原合成酶激酶-3β(GSK3β)在氧化應(yīng)激誘導(dǎo)肝細(xì)胞凋亡中的作用。方法以人肝HL-7702細(xì)胞為研究對象,H2O2/抗霉素A誘導(dǎo)細(xì)胞產(chǎn)生氧化應(yīng)激,建立肝細(xì)胞凋亡模型。SB216763為GSK3β特異性抑制劑,在H2O2/抗霉素A給藥前2 h干預(yù)。采用鈣黃綠素乙酰甲酯/碘化丙啶(PI)雙染色觀察細(xì)胞存活情況,采用annexinⅤ-FITC/PI聯(lián)合流式細(xì)胞術(shù)檢測細(xì)胞凋亡,同時(shí)對細(xì)胞培養(yǎng)上清進(jìn)行乳酸脫氫酶(LDH)檢測來評價(jià)細(xì)胞死亡程度;Western blot法檢測p-GSK3β、GSK3β、caspase-3、cleaved caspase-3、c-Jun氨基末端激酶(JNK)和細(xì)胞色素C(Cyt C)蛋白的表達(dá)。結(jié)果 H2O2/抗霉素A誘導(dǎo)的氧化應(yīng)激促進(jìn)了GSK3β活性增加,抑制GSK3β活性緩解了氧化應(yīng)激和由氧化應(yīng)激引起的細(xì)胞凋亡。與氧化應(yīng)激模型組相比,SB216763干預(yù)組中PI染色的細(xì)胞顯著減少,流式細(xì)胞術(shù)檢測細(xì)胞凋亡率降低,細(xì)胞培養(yǎng)上清中LDH顯著降低,Western blot法結(jié)果顯示干預(yù)組中cleaved caspase-3、JNK、Cyt C蛋白表達(dá)下降。結(jié)論 GSK3β是氧化應(yīng)激誘導(dǎo)細(xì)胞凋亡通路中的一種重要信號分子,抑制其活性可減輕氧化應(yīng)激而改善肝細(xì)胞凋亡。
[Abstract]:Objective to investigate the role of glycogen synthase kinase 3 尾 -GSK3 尾 in oxidative stress-induced hepatocyte apoptosis. Methods Hepatocyte apoptosis model, SB216763, was established by inducing oxidative stress in human hepatic HL-7702 cells induced by H _ 2O _ 2 / anti-mycin A, and SB216763 was used as a specific inhibitor of GSK3 尾. Two hours before the administration of H _ 2O _ 2 / anti-mycin A, the cell survival was observed by double staining of calcium xanthocyanin acetyl methyl ester / pyridine iodide, and apoptosis was detected by annexin 鈪，

本文編號：1564227