miR-125a-3p靶向作用P2RX7抑制糖尿病肾病小鼠肾纤维化发展

miR-125a-3p targeting P2RX7 inhibits the development of renal fibrosis in micewith diabetic nephropathy

摘要:

目的探究miR-125a-3p靶向作用P2RX7对糖尿病肾病(DN)小鼠肾纤维化发展的影响。 方法60只小鼠均分为对照组、模型组、miR-125a-3p NC组和miR-125a-3p mimic组,构建DN小鼠模型,腹腔注射miR-125a-3p NC或mimic。qRT-PCR检测肾组织miR-125a-3p表达,Western blotting检测P2RX7、Col-I、α-SMA蛋白表达,ELISA检测血肌酐(Cr)和血尿素氮(BUN),HE染色和Masson染色检测肾脏病理改变和胶原蛋白体积分数(CVF),双荧光素酶报告检测miR-125a-3p与P2RX7的靶向关系。 结果与对照组比较,模型组和miR-125a-3p NC组P2RX7、Cr、BUN和CVF、Col-I和α-SMA蛋白升高,miR-125a-3p表达降低(P＜0.05)；与模型组和miR-125a-3p NC组比较,miR-125a-3p mimic组上述趋势得到逆转(P＜0.05)。miR-125a-3p可与P2RX7靶向结合(P＜0.05)。 结论miR-125a-3p高表达靶向抑制P2RX7蛋白,缓解DN小鼠的肾纤维化,保护肾功能。

Abstract:

To explore the effect of miR-125a-3p targeting P2RX7on the development of renal fibrosis in diabetic nephropathy (DN) mice. Methods60mice were equally divided into control group, model group, miR-125a-3p NC group and miR-125a-3p mimic group. DN mouse model was induced, and the intraperitoneal injection of miR-125a-3p NC or mimic was performed. The relative expression level of miR-125a-3p was detected byreal-time quantitative polymerase chain reaction (qRT-PCR). The protein expression of P2RX7, Col-I, and α-SMA was detected by Western blotting. Serum levels of creatinine (Cr) and blood urea nitrogen (BUN) were detected by enzyme-linked immunosorbent assay (ELISA). The pathological changes of the kidney and tissue collagen volume fraction (CVF) were observed by HE staining and Masson staining, and the targeting relationship between miR-125a-3p and P2RX7 was detected by dual-luciferase reporter assay. ResultsCompared with control group, the P2RX7, Cr, BUN and CVF, Col-I and α-SMA proteins in the model group and the miR-125a-3p NC group were increased, and the expression of miR-125a-3p was decreased (P＜0.05). The above trend was reversed in miR-125a-3p mimic group compared with model group and miR-125a-3p NC group (all P＜0.05). miR-125a-3p could target and bind to P2RX7 (P＜0.05). ConclusionThe high expressionof miR-125a-3p can relieve DN-induced renal fibrosis and protect renal function via targeting P2RX7.