Tianjin Medical University, Tianjin 300070, China; Department of Minimal Invasive Surgery, Tianjin Nankai Hospital, Tianjin 300100, China.
Department of Minimal Invasive Surgery, Tianjin Nankai Hospital, Tianjin 300100, China.
Biomed Pharmacother. 2018 Apr;100:448-454. doi: 10.1016/j.biopha.2018.02.012. Epub 2018 Feb 22.
MicroRNAs (miRNAs) are involved in the pathological progression of various disease including type 2 diabetes (T2D). Chronic inflammation in adipose tissue is a cause of insulin resistance and T2D. MiR-17 palys an anti-inflammatory role in many biological processes. We hypothesized that miR-17 suppressed inflammatory macrophage that is related to insulin resistance in patients with T2D.
Macrophage migration and secretion of inflammatory cytokines including TNF-α, IL-6 and IL-1β were detected through transwell migration assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was tested by the radioactivity of tritium-labeled glucose in 3T3-L1 adipocytes. Dual luciferase reporter gene assay was employed to evaluate the interaction between miR-17 and 3'UTR of ASK1.
Our results showed that miR-17 inhibited macrophage infiltration and secretion of TNF-α, IL-6 and IL-1β. Moreover, insulin-stimulated glucose uptake of 3T3-L1 was suppressed by treatment with LPS-induced macrophage conditioned media (CM), whereas the opposite effect was showed after treatment with the CM of macrophages transfected with miR-17. Furthermore, we found that miR-17 directly prevented expression of ASK1 by binding to its 3'UTR.
miR-17 improved inflammation-induced insulin resistance by suppressing ASK1 expression in macrophages. These results indicated that miR-17 had an anti-diabetic acitivity by its anti-inflammation effect on macrophage.
微小 RNA（miRNA）参与包括 2 型糖尿病（T2D）在内的各种疾病的病理进展。脂肪组织中的慢性炎症是胰岛素抵抗和 T2D 的原因。miR-17 在许多生物过程中发挥抗炎作用。我们假设 miR-17 抑制与 T2D 患者胰岛素抵抗相关的炎症性巨噬细胞。
通过 Transwell 迁移测定和酶联免疫吸附试验分别检测巨噬细胞迁移和炎症细胞因子（包括 TNF-α、IL-6 和 IL-1β）的分泌。通过 3T3-L1 脂肪细胞中氚标记葡萄糖的放射性来测试胰岛素刺激的葡萄糖摄取。双荧光素酶报告基因测定用于评估 miR-17 与 ASK1 的 3'UTR 之间的相互作用。
我们的结果表明，miR-17 抑制巨噬细胞浸润和 TNF-α、IL-6 和 IL-1β 的分泌。此外，用 LPS 诱导的巨噬细胞条件培养基（CM）处理会抑制 3T3-L1 的胰岛素刺激的葡萄糖摄取，而用转染 miR-17 的巨噬细胞的 CM 处理则显示出相反的效果。此外，我们发现 miR-17 通过与其 3'UTR 结合直接阻止 ASK1 的表达。
miR-17 通过抑制巨噬细胞中 ASK1 的表达改善了炎症诱导的胰岛素抵抗。这些结果表明，miR-17 通过其对巨噬细胞的抗炎作用具有抗糖尿病活性。
