丙型肝炎病毒核心蛋白对低氧诱导因子-1α和血管内皮生长因子表达的影响

Effects of HCV core protein on the expression of hypoxia-inducible factor 1a and vascular endothelial growth facto

目的 探讨HCV核心蛋白对低氧诱导因子1α【（HIF-1α）和血管内皮生长因子（VEGF）表达的影响。方法将HCV核心蛋白基因的真核表达载体flag2B—core和HIF-1αsiRNA转染Huh7．5．1细胞；采用RT-PCR和Westernblot法分别检0n,0HIF～1α、VEGF在mRNA和蛋白水平的表达变化，采用酶联免疫吸附试验检测细胞上清液中VEGF的含量。对各组间数据进行t检验。结果Huh7．5．1细胞转染flag2B-core后，HIF-1α和VEGF的mRNA和蛋白表达水平升高，细胞上清液中VEGF含量明显高于对照组[（654．5±43．7）pg／ml与（365．9±26．8）pg／ml，t=653．1％，P〈0．01）]；Huh7．5．1细胞共转染flag2B—core和HIF-1asiRNA后，HIF-1a和VEGF的mRNA和蛋白表达水平降低，细胞上清液VEGF含量明显低于对照组[（389．2±29．6）pg／ml与（768．8±47．3）pg／ml，t=1330．22，P〈0．01）。结论HCV核心蛋白能够上调HIE-1α和VEGF的表达；HCV可能通过核心蛋白来调节HIF-1α和VEGF的表达。

Objective To investigate the effect of core protein of hepatitis C virus （HCV） on the expression of hypoxia-inducible factor-1α （HIF-1α and vascular endothelial growth factor （VEGF）. Methods HuhT.5.1 cells were transfected with plasmid flag2B-core carrying HCV core gene, expression of HIF-1α and VEGF were measured by reverse transcription-polymorphism chain reaction （RT-PCR） and western blot. Enzyme link immunobsorbent assay （ELISA） were used to detect the level of VEGF in the supernatants. Results The expression of HIF-1α and VEGF mRNA and protein were upregulated after flag2B-core was transfected into HuhT.5.1 cells, and VEGF level in the supematant was significant elevated as compared to controls [（654.5±43.7） pg/ml vs （365.9±26.8） pg/ml, t = 653.1%, P〈0.01]. The expression of HIF-1a and VEGF mRNA and protein were downregulated after flag2B-core and HIF-1a siRNA were co-transfected into Huh7.5.1 cells, and VEGF level in the supernatant was significanty reduced as compared to controls [（389.2±29.6） pg/ml vs （768.8±47.3）pg/ml, t = 1330.22, P〈0.01]. Conclusions HCV core protein enhances the expression of HIF-1a and VEGF. HCV may regulate the expression of HIF-1a and VEGF via the core protein.