乙醇诱导HepG_2细胞凋亡模型的建立及Bcl-2对抗作用

Resistance of Bcl-2 adenovirus vector to HepG_2 cell apoptosis induced by ethanol

目的研究 Ｂｃｌ－２家族蛋白在乙醇引发肝细胞凋亡时的作用。方法以 ＴＵＮＥＬ法、 ＤＮＡ梯度检测乙醇诱导ＨｅｐＧ_２肝细胞凋亡，以免疫组织化学法检测细胞中Ｂｃｌ－２、Ｂａｘ、Ｂａｋ等蛋白表达情况，并以Ｂｃｌ－２腺病毒感染细胞观察其抗凋亡作用。结果体积分数 ０．２％、 １．０％乙醇处理 ２４ ｈ对 ＨｅＰＧ_２细胞无明显细胞毒性作用；３．０％组则有明显的细胞毒性作用，伴有典型的凋亡特征。免疫组织化学发现Ｂａｘ、Ｂａｋ蛋白在３．０％组显著增加；Ｂｃｌ－２腺病毒载体可部分抑制凋亡。结论 Ｂａｘ、Ｂａｋ蛋白加强表达在乙醇诱导ＨｅｏＧ_２细胞凋亡中起一定作用； Ｂｃｌ－２腺病毒载体可有效抑制乙醇诱导的凋亡。

Objective To evaluate the role of Bcl-2 family proteins in hepatocytic apoptosis caused by ethanol. Methods We induced hepatocytic apoptosis in HepG_2 cells with ethanol and detected the apoptosis with TUNEL assay. Bcl-2, Bax, and Bak protein expression in the hepatocytes was determined by immunohistochemistry method. We also observed the expression of Bcl-2 protein in HepG_2 cells infected with Bcl-2 adenovirus vector and its protection against hepatocytic apoptosis caused by ethanol. Results No obvious cytotoxicity was noted in 0.2% and l% ethanol treatment group, while it developed in 3% ethanol treatment group and accompanied by a marked expression of Bax, Bak proteins. Conclusion The overexpression of Bax, Bak proteins may play a role in HepG_2 cell apoptosis induced by ethanol and can be blocked effectively by Bcl-2 adenovirus vector.