Protection of rat hepatocytes from apoptosis by inhibition of c-Jun N-terminal kinase
Protection of rat hepatocytes from apoptosis by inhibition of c-Jun N-terminal kinase
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DOI：
10.1067/msy.2003.237
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发表时间：
2003-08-01
期刊：
影响因子：
3.8
通讯作者：
Geller, DA
中科院分区：
文献类型：
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作者：
Marderstein, EL;Bucher, B;Geller, DA
背景肝缺血/再灌注损伤后发生细胞凋亡和c-Jun N-末端激酶（JNK）激活。在其他细胞类型中，JNK激活被证明是细胞凋亡所需的。本研究验证了JNK参与肝细胞凋亡和抑制JNK活性提高细胞活力的假设。从Sprague道利大鼠收获大鼠肝细胞，并用JNK抑制剂SP 600125预处理。随后，他们暴露于由胆盐甘氨鹅去氧胆酸（GCDC）或肿瘤坏死因子（TNF）-α和放线菌素D组成的凋亡刺激。Western blotting证实SP 600125对JNK有特异性抑制作用。抑制JWK导致改善的生存能力测量结晶紫，减少原位DNA缺口末端标记阳性，并减少裂解的聚（ADP-核糖）聚合酶和caspase-3。TNF-α和放线菌素D诱导细胞凋亡，上调p53，下调抗凋亡蛋白X-连锁凋亡抑制蛋白的表达。这些作用可被JNK抑制剂所消除。这些数据表明，JNK活性的药理学抑制通过维持抗凋亡蛋白的表达来减少胆盐或TNF α诱导的凋亡。结果表明，JNK是凋亡信号级联的重要组成部分，并建议在某些肝脏疾病的可能的治疗策略。
Background. Apoptotic cell death and c-Jun N-terminal kinase (JNK) activation occur after hepatic ischemia/reperfusion injury. In other cell types, JNK activation was shown to be required for apoptosis. This study tested the hypotheses that JNK contributes to hepatocellular apoptosis, and that inhibition of JNK activity improves cell viability.Methods. Rat hepatocytes were harvested from Sprague Dawley rats and pretreated with SP600125, a JNK inhibitor. Subsequently, they were exposed to apoptotic stimuli consisting of either the bile salt glycochenodeoxycholic acid (GCDC) or tumor necrosis factor (TNF)-alpha and actinomycin D.Results. Western blotting demonstrated specific inhibition of JNK by SP600125. Inhibition of JWK resulted in improved viability measured with crystal violet, decreased in situ DNA nick end labeling positivity, and decreased cleavage of poly (ADP-ribose) polymerase and caspase-3. TNF-a and actinomycin D induced apoptosis, upregulated p53, and downregulated expression of the anti-apoptotic protein X-linked inhibitor of apoptosis protein. These effects were abrogated by JNK inhibition.Conclusion. These data show that pharmacologic inhibition of JNK activity reduces bile salt or TNFalpha-induced apoptosis by maintaining expression of anti-apoptotic proteins. The results indicate that JNK is an important component of the apoptosis signaling cascade and suggest a possible therapeutic strategy in certain liver disorders.