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

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。

Aim To explore the effect of miR-125a-3p targeting P2RX7on the development of renal fibrosis in diabetic nephropathy(DN)mice.Methods:60mice 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 quanti-tative polymerase chain reaction(qRT-PCR).The protein expression of P2RX7,Col-I,andα-SMA was detected by Western blotting.Serum levels of creatinine(Cr)and blood urea nitrogen(BUN)were detected by enzyme-linked immu-nosorbent assay(ELISA).The pathological changes of the kidney and tissue collagen volume fraction(CVF)were ob-served by HE staining and Masson staining,and the targeting relationship between miR-125a-3p and P2RX7 was detected by dual-luciferase reporter assay.Results:Compared with control group,the P2RX7,Cr,BUN and CVF,Col-I andα-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).Conclusion The high expressionof miR-125a-3p can relieve DN-induced renal fibrosis and protect renal function via targeting P2RX7.