IL-32对HepG22.15细胞凋亡的影响

Effect of Interleukin-32 on the Apoptosis of HepG2 2.15 Cells

目的研究白细胞介素-32（IL-32）对乙型肝炎病毒（HBV）感染的肝细胞（HepG2 2.15细胞,插入HBV全基因组,持续表达）凋亡的影响。方法将不同浓度的IL-32（0~0.4μg/mL）蛋白质与HepG2细胞、HepG2 2.15细胞共培养,48h后用MTT检测细胞增殖。收集与IL-32共培养48h的HepG2细胞、HepG2 2.15细胞,加入AnnexinⅤ和PI,室温避光孵育15min,流式细胞仪检测细胞凋亡。将IL-32（0.2μg/mL）与HepG2细胞、HepG2 2.15细胞共培养,分别于0、12、24h后收集细胞,Human Active Caspase-3Immunoassay检测试剂盒检测活化的Caspase-3表达水平,Caspase-3Colorimetric检测试剂盒检测Caspase-3酶活性。结果IL-32抑制HepG2细胞、HepG2 2.15增殖,诱导HepG2细胞、HepG2 2.15细胞凋亡,且随着IL-32浓度的增高,细胞凋亡水平亦增高。IL-32诱导HepG2细胞、HepG22.15细胞活化的Caspase-3表达水平及Caspase-3酶活性均增加。结论 IL-32可能通过活化Caspase-3通路诱导HBV感染的肝细胞凋亡而发挥抗病毒作用,同时亦参与慢性乙型病毒性肝炎肝损伤的发生。

Objective To investigate the effect of interleukin-32（IL-32）on the apoptosis of HepG2 2.15 cells in which the whole genome of HBV was inserted and persistently expressed.Methods HepG2 cells or HepG2 2.15 cells were treated with different concentrations of IL-32（0-0.4μg/mL）.Forty-eight hours later,cell proliferation was assayed by MTT method.Cell apoptosis was assayed by flow cytometry（FCM）after treatment of both cells with IL-32 for 48 h,and then with annexin Ⅴ and PI at room temperature in the dark for 15 min.Subsequently,HepG2 cells or HepG2 2.15 cells were co-cultured with IL-32 at 0.2μg/mL.They were collected at 0,12,24 h,respectively.Activated caspase-3 expression levels were assayed by using Human Active Caspase-3 Immunoassay Kit.Caspase-3 enzyme activities were detected by Caspase-3 Colorimetric Assay Kit.Results IL-32 could inhibit the proliferation of HepG2 cells and HepG2 2.15 cells,and induce their apoptosis in a dose-dependent man-ner.Activated caspase-3 expression levels and caspase-3 enzyme activities were increased in both HepG2 and HepG2 2.15 cells co-cultured with IL-32.Conclusion IL-32 play an anti-viral role by activating the caspase-3 pathway and inducing the apoptosis of HBV-infected hepatocytes,and it is involved in the liver damage of chronic hepatitis B at the same time.