Department of Endocrinology and Metabolism, The First People's Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan, China.
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, Yunnan, China.
Anal Cell Pathol (Amst). 2023 Jun 17;2023:5350999. doi: 10.1155/2023/5350999. eCollection 2023.
Diabetes is one of the most common diseases and major public health burdens worldwide. Type 2 diabetes mellitus (T2DM) is associated with chronic hepatitis C virus (HCV) infection, and lncRNAs play an important role in HCV-induced T2DM. We aimed to explore the effect of lncRNA AC040162.3 on HCV-induced T2DM.
HCV was used to infect MIN6 cells to establish an in vitro model. HCV copy number and miRNA expression were detected by Real Time Quantitative PCR (RT-qPCR). Enzyme-Linked Immunosorbent Assay (ELISA) was used to detect the secretion of insulin, and methyl thiazolyl tetrazolium (MTT) was applied to analyze cell viability. Apoptosis was analyzed by Western blotting and flow cytometry. In addition, Western blotting and TdT-mediated dUTP Nick End Labeling (TUNEL) were used to analyze pyroptosis. Luciferase reporter assays were used to investigate the targeting relationship.
The expression of LncRNA AC040162.3 and NLRP3 was markedly increased in HCV-T2DM, while the expression of miR-223-3p was remarkably inhibited. In vitro experiments demonstrated that lncRNA AC040162.3 silencing or miR-223-3p overexpression remarkably alleviated HCV-induced T2DM deterioration by inhibiting cell apoptosis and pyroptosis and enhancing cell viability. We then demonstrated that silencing lncRNA AC040162.3 promoted the expression of miR-223-3p and that miR-223-3p bound to lncRNA AC040162.3 and the NLRP3 binding site. In addition, the protective effects of LncRNA AC040162.3 silencing in HCV-infected MIN6 cells were reversed by overexpression of NLRP3 or silencing of miR-223-3p.
Silencing of lncRNA AC040162.3 alleviates the process of HCV-induced T2DM by governing the miR-223-3p/NLRP3 axis.
糖尿病是全球最常见的疾病和主要公共卫生负担之一。2 型糖尿病（T2DM）与慢性丙型肝炎病毒（HCV）感染有关，lncRNAs 在 HCV 诱导的 T2DM 中发挥重要作用。我们旨在探讨 lncRNA AC040162.3 对 HCV 诱导的 T2DM 的影响。
用 HCV 感染 MIN6 细胞建立体外模型。采用实时定量 PCR（RT-qPCR）检测 HCV 拷贝数和 miRNA 表达。酶联免疫吸附试验（ELISA）检测胰岛素分泌，甲基噻唑基四唑（MTT）分析细胞活力。Western blot 和流式细胞术分析细胞凋亡。此外，Western blot 和末端转移酶介导的 dUTP 缺口末端标记（TUNEL）分析细胞焦亡。荧光素酶报告实验用于研究靶向关系。
在 HCV-T2DM 中，LncRNA AC040162.3 和 NLRP3 的表达明显增加，而 miR-223-3p 的表达明显受到抑制。体外实验表明，沉默 LncRNA AC040162.3 或过表达 miR-223-3p 可显著减轻 HCV 诱导的 T2DM 恶化，抑制细胞凋亡和焦亡，增强细胞活力。然后，我们证明沉默 LncRNA AC040162.3 可促进 miR-223-3p 的表达，并且 miR-223-3p 结合到 LncRNA AC040162.3 和 NLRP3 结合位点上。此外，过表达 NLRP3 或沉默 miR-223-3p 可逆转沉默 LncRNA AC040162.3 在 HCV 感染的 MIN6 细胞中的保护作用。
沉默 LncRNA AC040162.3 通过调控 miR-223-3p/NLRP3 轴减轻 HCV 诱导的 T2DM 进程。
