Bcl-2腺病毒载体抗肿瘤坏死因子α及氨基半乳糖诱导小鼠肝细胞凋亡

The effect of Bcl-2 adenovirus vector against murine hepatocyte apoptosis caused by tumor necrosis factor α and D-galactosamine

目的 探讨Ｂｃｌ－２家族蛋白在肿瘤坏死因子α （ＴＮＦα）致肝损伤及肝细胞凋亡中的作用。方法以ＴＮＦα联合Ｄ－氨基半乳糖诱发小鼠肝损伤，以免疫组织化学法检测Ｂａｘ、Ｂａｋ蛋白在鼠肝组织中的表达情况；并以Ｂｃｌ－２腺病毒载体感染肝细胞，观察其抗肝细胞周亡作用。结果ＴＮＦα可引起严重的肝损伤并有广泛的肝细胞凋亡，伴Ｂａｘ、 Ｂａｋ蛋白在肝细胞中表达增强；Ｂｃｌ－２腺病毒感染可使肝损伤小鼠ＡＬＴ水平由（１３７２．９ ± ２５１．４）Ｕ／Ｌ下降至（７９６．５±７８．７）Ｕ／Ｌ，统计分析差异有显著意义（Ｐ＜０．０００５）。结论ＦＮＦα诱导肝细胞凋亡可能与其诱导Ｂａｘ、 Ｂａｋ蛋白在肝细胞中表达增强有关； Ｂｃｌ－２腺病毒载体可在小鼠肝细胞中持续表达至少１个月并可部分抵抗ＴＮＦα诱发的肝细胞凋亡。

Objective To evaluate the role of Bcl-2 family proteins in hepatic apoptosis caused by TNF α and Dgalactosamine. Methods We induced mouse liver injury with TNF α and D-galactosamine, and detected hepatic apoptosis, the expression of Bcl-2, Bax, and Bak proteins on hepatocytes by using TUNEL or immunohistochemistry, respectively. We also observed the expression of Bcl-2 protein on hepatocytes infected with Bcl-2 adenovirus vector and its protection against hepatocyte apoptosis. Results Hepatocyte apoptosis was induced in BalB/c mice pretreated with TNF α plus Dgalactosamine, accompanying the enhanced expression of Bax, Bak proteins in hepatocytes. Bcl-2 protein was expressed in murine hepatocytes and lasted at least 1 month after injection of Bcl-2 adenovirus vector, which also lowered ALT level from (1372.9 ± 251 .4) U/L to (796.5 ± 78.7 )U/L and reduced hepatocyte apoptosis caused by TNF α and D-galactosamine. Conclusions The enhanced expression of Bax, Bak proteins may play a role in hepatocyte apoptosis induced by TNF α and D-galactosamine. D-galactosamine adenovirus vector can partially reduce hepatocyte apoptosis induced by TNF α and D-galactosamine.