Department of Endocrinology and Metabolism, Fourth Affiliated Hospital, China Medical University, Shenyang, Liaoning, China.
Department of Endocrinology and Metabolism, Fourth Affiliated Hospital, China Medical University, Shenyang, Liaoning, China.
Gene. 2019 May 25;698:150-156. doi: 10.1016/j.gene.2019.02.047. Epub 2019 Mar 6.
Investigating the expression of the lnc RNAs screened above between normal and insulin resistant 3T3-L1 adipocytes. Addressing the mechanism underlying the regulation of inflammation response by lnc TINCR.
3T3-L1 preadipocytes were induced to differentiate into mature adipocytes. Oil red O staining was used to find the fat droplets in mature adipocytes. Mature adipocytes were randomized to normal control group and Tri-DAP (NOD1 ligand) group. After the establishment of insulin resistance model, we used deep RNA sequencing(RNA-Seq) to identify lncRNAs that are regulated during NODI activation in mouse adipocytes. Real-time PCR was used to analyze the expression of lnc TINCR, proinflammatory IL-6, TNF-α, Cxcl1 and RIPK2 in the presence or absence of Tri-DAP(10 μg/ml). We employed siRNA against lnc TINCR to confirm its effects in inflammatory response.
Deep RNA sequencing identified 81 lncRNAs and 167 coding genes that were significantly up-related while 78 lncRNAs and 82 coding genes that were significantly down-related greater than twofold during NOD1 activation in adipocytes. We discovered that lnc TINCR, termed lnc TINCR(Tri-DAP-inducible non-protein coding RNA) is greatly upregulated in Tri-DAP activated adipocytes. Moreover knockdown of lnc TINCR dampens the proinflammatory response (P < 0.05; in adipocytes).
lnc TINCR is a positive regulator of inflammation-induced insulin resistance presumably via activation of NOD1 signaling pathways.
研究上述筛选出的长非编码 RNA（lncRNA）在正常和胰岛素抵抗 3T3-L1 脂肪细胞之间的表达。探讨 lncTINCR 调节炎症反应的机制。
将 3T3-L1 前脂肪细胞诱导分化为成熟脂肪细胞。油红 O 染色法寻找成熟脂肪细胞中的脂肪滴。将成熟脂肪细胞随机分为正常对照组和三-DAP（NOD1 配体）组。在建立胰岛素抵抗模型后，我们使用深度 RNA 测序（RNA-Seq）鉴定在 NOD1 激活过程中在小鼠脂肪细胞中受到调节的 lncRNA。实时 PCR 用于分析在存在或不存在三-DAP（10μg/ml）的情况下，lncTINCR、促炎细胞因子 IL-6、TNF-α、Cxcl1 和 RIPK2 的表达。我们采用针对 lncTINCR 的 siRNA 来确认其在炎症反应中的作用。
深度 RNA 测序鉴定出 81 个 lncRNA 和 167 个编码基因在 NOD1 激活过程中显著上调，而 78 个 lncRNA 和 82 个编码基因显著下调两倍以上。我们发现 lncTINCR（称为 lncTINCR（三-DAP 诱导非蛋白编码 RNA））在三-DAP 激活的脂肪细胞中显著上调。此外，lncTINCR 的敲低抑制了促炎反应（P<0.05；在脂肪细胞中）。
lncTINCR 是炎症诱导的胰岛素抵抗的正调节剂，可能通过激活 NOD1 信号通路。
