Centre of Excellence in Molecular Biology, University of Punjab, Lahore, 54000, Lahore, Pakistan.
Department of Mathematics, Division of Science and Technology, University of Education, Lahore, 54000, Pakistan.
Sci Rep. 2019 Apr 16;9(1):6150. doi: 10.1038/s41598-019-42602-2.
HCV genes interfere with host cellular genes and play crucial role in pathogenesis. The mechanism under which HCV genes induce insulin resistance is not much clear. This study is aimed to examine the role of HCV NS5A in inducing insulin resistance by examining its affect in the phosphorylation level of AKT/PKB. In the present study, HepG2 cells were transfected with HCV NS5A and after 24 hours of transfection, protein was extracted from cells that were pre induced with insulin at three different time intervals i.e. 1hour, 2 hours and 3hours. Dot Blot analysis was performed to study the phosphorylation level of AKT. Results showed that there is clear upregulation of serine 473 phosphorylation level of AKT in NS5A transfected cells as compared with control (without NS5A). In conclusion, upregulation of serine 473 phosphorylation by NS5A of HCV genotype 3a suggests that this gene impairs the normal insulin AKT/PKB signaling pathway that leads towards insulin resistance and Type 2 diabetes mellitus. Therefore, HCV non-structural protein NS5A should be considered as promising candidate to be studied in detail for HCV induced insulin resistance and should be regarded as a therapeutically important target for the prevention of chronic liver diseases.
丙型肝炎病毒基因干扰宿主细胞基因，并在发病机制中发挥关键作用。丙型肝炎病毒基因诱导胰岛素抵抗的机制尚不清楚。本研究旨在通过研究 HCV NS5A 对 AKT/PKB 磷酸化水平的影响，来检验其诱导胰岛素抵抗的作用。在本研究中，将 HCV NS5A 转染 HepG2 细胞，转染 24 小时后，用胰岛素在三个不同时间点（即 1 小时、2 小时和 3 小时）预先诱导细胞，从细胞中提取蛋白质。采用斑点印迹分析研究 AKT 的磷酸化水平。结果表明，与对照（无 NS5A）相比，NS5A 转染细胞中 AKT 的丝氨酸 473 磷酸化水平明显上调。结论：丙型肝炎病毒 3a 型 NS5A 的上调可导致丝氨酸 473 磷酸化，表明该基因破坏了正常的胰岛素 AKT/PKB 信号通路，导致胰岛素抵抗和 2 型糖尿病。因此，丙型肝炎病毒非结构蛋白 NS5A 应被视为研究丙型肝炎病毒诱导胰岛素抵抗的有前途的候选物，并应被视为预防慢性肝病的重要治疗靶点。
