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目的探讨脂多糖(LPS)、巨噬细胞表达肿瘤坏死因子相关凋亡诱导配体(TRAIL)和肝细胞凋亡的相互关系。方法LPS刺激巨噬细胞后,以流式细胞仪测定细胞表面的TRAIL表达,用酶联免疫吸附法(ELISA)法检测培养上清中可溶性TRAIL的表达;以(?)铬释放试验测定可溶性TRAIL和膜性TRAIL 对HepG2细胞的毒性作用,并用Annexin V染色法验证凋亡细胞的产生。结果经100 ng/ml LPS刺激后, 只有9.8%的巨噬细胞有TRAIL的表达,而培养上清中可溶性TRAIL达(67.4±5.1)ng/ml,有明显增加。巨噬细胞培养上清液中可溶性TRAIL能溶解HepG2细胞,经Annexin V染色法证实为细胞凋亡,此作用可被特异性TRAIL抗体阻断。结论LPS能增加巨噬细胞可溶性TRAIL表达,并诱导肝细胞凋亡,提示TRAIL在病毒性肝炎发病机制中有重要作用。
Objective To investigate the relationship between the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and hepatocyte apoptosis in lipopolysaccharide (LPS) and macrophages. Methods After LPS stimulated macrophages, the expression of TRAIL on the cell surface was determined by flow cytometry. The expression of soluble TRAIL was detected by enzyme linked immunosorbent assay (ELISA). The soluble TRAIL And membrane-bound TRAIL on HepG2 cells. Annexin V staining was used to verify the apoptotic cells. Results After stimulated with 100 ng / ml LPS, only 9.8% of the macrophages had TRAIL expression, while the soluble TRAIL in the culture supernatant was (67.4 ± 5.1) ng / ml, which was significantly increased. Soluble TRAIL in macrophage culture supernatant could dissolve HepG2 cells, which was confirmed by Annexin V staining as apoptosis. This effect could be blocked by specific TRAIL antibody. Conclusion LPS can increase the soluble TRAIL expression in macrophages and induce the apoptosis of hepatocytes, suggesting that TRAIL plays an important role in the pathogenesis of viral hepatitis.