Molecular Endocrinology, Functional Genomics and Systems Biology Laboratory, Animal Biochemistry Division, National Dairy Research Institute, Karnal, 132001, Haryana, India.
Molecular Endocrinology, Functional Genomics and Systems Biology Laboratory, Animal Biochemistry Division, National Dairy Research Institute, Karnal, 132001, Haryana, India.
Mol Immunol. 2020 Mar;119:154-158. doi: 10.1016/j.molimm.2020.01.019. Epub 2020 Feb 6.
A genome-wide profiling of microRNA (miRNA) in endotoxin tolerant buffalo granulosa cells identified miR-326 amongst top-10 upregulated miRNAs. In this study, we have elucidated the role of miR-326 in granulosa cells in vitro. In-silico analysis revealed that miR-326 have binding site for 3'UTR of TLR-4 (Toll-like receptor 4). Transfection experiments showed that there was a significant inhibition of TLR-4 when miR-326 mimic was transfected to buffalo granulosa cells. Furthermore, when miR-326 transfected granulosa cells were exposed to LPS, followed by expression analysis of TLR-4 complex genes (TLR-4 and MyD88) and pro-inflammatory cytokines, we found a decreased expression of both TLR-4 complex and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β). We also found that the expression of anti-inflammatory (IL-10) gene was upregulated. To the best of our knowledge, this is the first study that showed the regulation of TLR-4 using miR-326. The present findings on regulation of TLR-4 are important and would help in understanding innate immunity regulation under different patho-physiology.
利用内毒素耐受水牛颗粒细胞进行全基因组 miRNA 分析，确定了 miR-326 在 10 个上调 miRNA 中排名靠前。在这项研究中，我们阐明了 miR-326 在体外颗粒细胞中的作用。计算机分析显示，miR-326 有 TLR-4（Toll 样受体 4）3'UTR 的结合位点。转染实验表明，当 miR-326 模拟物转染到水牛颗粒细胞时，TLR-4 受到显著抑制。此外，当 miR-326 转染的颗粒细胞暴露于 LPS 后，对 TLR-4 复合物基因（TLR-4 和 MyD88）和促炎细胞因子的表达分析发现，TLR-4 复合物和促炎细胞因子（TNF-α、IL-6 和 IL-1β）的表达均降低。我们还发现抗炎（IL-10）基因的表达上调。据我们所知，这是第一项表明 miR-326 调节 TLR-4 的研究。TLR-4 调节的这一发现非常重要，将有助于理解不同病理生理学下的固有免疫调节。
