Chandipura virus (CHPV) is a negative single-stranded RNA virus of the Rhabdoviridae family. CHPV infection has been reported in Central and Western India. CHPV causes acute encephalitis with a case fatality rate of 70 % and mostly affects children below 15 years of age. CHPV infection in brain leads to neuronal apoptosis and activation of the microglial cells. The microRNAs (miRNAs) are small endogenous non-coding RNA that regulate the gene expression. Viral infections perturb the expression pattern of cellular miRNAs, which may in turn affect the expression pattern of downstream genes. This study aims to investigate hsa-miR-21-5p mediated regulation of PTEN, AKT, NF-ĸBp65, IL-6, TNF-α, and IL-1β, in human microglial cells during CHPV infection. To understand the role of hsa-miR-21-5p in CHPV infection, the human microglial cells were infected with CHPV (MOI-0.1). Real-time PCR, western blotting, Luciferase assay, over-expression and knockdown techniques were used to understand the role of hsa-miR-21-5p in the regulation of PTEN, AKT and, NF-ĸBp65, IL-6, TNF-α, and IL-1β in this study. The hsa-miR-21-5p was found to be upregulated during CHPV infection in human microglial cells. This led to the downregulation of PTEN which promoted the phosphorylation of AKT and NF-ĸBp65. Over-expression of hsa-miR-21-5p led to the decreased expression of PTEN and promoted further phosphorylation of AKT and NF-ĸBp65 in human microglial cells. However, the inhibition of hsa-miR-21-5p using hsa-miR-21-5p inhibitor restored the expression. This study supports the role of hsa-miR-21-5p in the regulation of pro-inflammatory genes in CHPV infected human microglial cells.

中文翻译：

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钱迪普拉病毒失调 hsa-miR-21-5p 的表达以激活人小胶质细胞中的 NF-κB

钱迪普拉病毒 (CHPV) 是弹状病毒科的一种阴性单链 RNA 病毒。CHPV感染在印度中部和西部已有报道。CHPV 引起急性脑炎，病死率为 70%，主要影响 15 岁以下的儿童。脑中的 CHPV 感染导致神经元凋亡和小胶质细胞活化。微小RNA (miRNA) 是调节基因表达的小的内源性非编码RNA。病毒感染会扰乱细胞 miRNA 的表达模式，进而影响下游基因的表达模式。本研究旨在研究在 CHPV 感染期间人小胶质细胞中 hsa-miR-21-5p 介导的 PTEN、AKT、NF-ĸBp65、IL-6、TNF-α 和 IL-1β 的调节。为了了解 hsa-miR-21-5p 在 CHPV 感染中的作用，人小胶质细胞感染了 CHPV (MOI-0.1)。实时 PCR、蛋白质印迹、荧光素酶测定、过表达和敲低技术用于了解 hsa-miR-21-5p 在 PTEN、AKT 和 NF-ĸBp65、IL-6、TNF-本研究中的α和IL-1β。发现 hsa-miR-21-5p 在人小胶质细胞的 CHPV 感染期间上调。这导致 PTEN 的下调，从而促进了 AKT 和 NF-ĸBp65 的磷酸化。hsa-miR-21-5p 的过表达导致 PTEN 的表达降低并促进人小胶质细胞中 AKT 和 NF-ĸBp65 的进一步磷酸化。然而，使用 hsa-miR-21-5p 抑制剂抑制 hsa-miR-21-5p 恢复了表达。