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目的:探讨microRNA-30(miR-30)缓解血管紧张素Ⅱ(AngⅡ)所致足细胞损伤的分子机制。方法:(1)通过皮下埋置渗透压泵,给予小鼠AngⅡ[1 000 ng/(kg·min)×28d]构建肾损伤模型。利用原位杂交及qRT-PCR技术检测小鼠肾小球中miR-30s的表达,运用免疫组化及蛋白印迹检测calcineurin信号重要分子细胞瞬时受体电位阳离子通道蛋白6(TRPC6),钙调磷酸酶(PPP3CA、PPP3CB、PPP3R1)及活化T细胞核因子(NFATC3)表达。(2)给予miR-30s慢病毒尾静脉注射,观察其对AngⅡ诱导损伤模型的肾脏保护作用。(3)体外研究观察miR-30a高表达能否抑制AngⅡ(10-6mol/L)诱导的足细胞骨架损伤和足细胞凋亡。结果:(1)AngⅡ诱导小鼠肾小球中miR-30s水平下调,并上调TRPC6,激活calcium/calcineurin信号通路;(2)高表达的miR-30s能够抑制AngⅡ导致的calcium/calcineurin信号活化从而缓解AngⅡ诱导的肾小球损伤;(3)miR-30s能够逆转AngⅡ所致足细胞骨架损伤及抑制其凋亡过程。结论:miR-30s具有抑制AngⅡ诱导的足细胞损害作用,其机制可能与抑制calcium/calcineurin信号通路有关。
AIM: To investigate the molecular mechanism of microRNA-30 (miR-30) in relieving podocyte injury induced by angiotensin Ⅱ (AngⅡ). Methods: (1) A rat model of renal injury was established by subcutaneously infiltrating the osmotic pressure pump with Ang Ⅱ [1 000 ng / (kg · min) × 28 d]. The expression of miR-30s in glomeruli of mice was detected by in situ hybridization and qRT-PCR. The expressions of TRPC6, TRAP6 and calcineurin were detected by immunohistochemistry and Western blot. (PPP3CA, PPP3CB, PPP3R1) and activated T cell nuclear factor (NFATC3) expression. (2) The miR-30s lentivirus was injected into tail vein to observe its protective effect on AngⅡinduced injury model. (3) In vitro study to observe whether miR-30a overexpression can inhibit AngⅡ (10-6mol / L) induced podocyte cytoskeleton injury and podocyte apoptosis. Results: (1) miR-30s was down-regulated in glomeruli of AngⅡ-induced mice and TRPC6 was upregulated to activate calcium / calcineurin signaling pathway; (2) miR-30s overexpression inhibited the activation of calcium / calcineurin signaling induced by AngⅡ (3) miR-30s can reverse AngⅡ-induced podocyte cytoskeleton damage and inhibit its apoptosis. Conclusion: miR-30s can inhibit AngⅡ-induced podocyte injury, which may be related to the inhibition of calcium / calcineurin signaling pathway.