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.

中文翻译：

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miR-326的上调调节水牛颗粒细胞中靶向TLR-4的促炎细胞因子。

内毒素耐受的水牛颗粒细胞中的microRNA（miRNA）的全基因组概况分析在前10个上调的miRNA中鉴定了miR-326。在这项研究中，我们阐明了miR-326在体外颗粒细胞中的作用。电子计算机分析表明，miR-326具有与TLR-4（Toll样受体4）的3'UTR结合的位点。转染实验表明，将miR-326模拟物转染到水牛颗粒细胞中时，TLR-4具有明显的抑制作用。此外，当将miR-326转染的颗粒细胞暴露于LPS，然后对TLR-4复杂基因（TLR-4和MyD88）和促炎细胞因子进行表达分析时，我们发现TLR-4复杂基因和pro炎症细胞因子（TNF-α，IL-6和IL-1β）。我们还发现抗炎（IL-10）基因的表达上调。据我们所知，这是第一项显示使用miR-326调节TLR-4的研究。目前关于TLR-4调控的发现很重要，有助于理解不同病理生理学下的先天免疫调控。