微小RNA-302c在高糖下对人肾小球系膜细胞的影响

Effect of MiR-302c on Human Glomerular Mesangial Cells in High Glucose Environment

目的探讨微小RNA-302c(miR-302c)在高糖环境下对人肾小球系膜细胞功能的影响。方法采用双荧光素酶报告实验验证miR-302c是否参与TIMP3表达的调控;在高糖环境下培养人肾小球系膜细胞,采用实时荧光定量PCR(qRT-PCR)法检测细胞miR-302c以及TIMP3 mRNA相对表达量,检测肾小球系膜细胞的存活数,采用qRT-PCR方法检测细胞炎症因子IL-6以及IL-8 mRNA的表达。结果经双荧光素酶报告基因实验证实TIMP3是miR-302c的直接靶基因;在不同糖浓度下培养人肾小球系膜细胞,结果发现,与正常糖培养相比较,高糖处理下的人肾小球系膜细胞的miR-302c表达显著性增加,TIMP3 mRNA表达显著性下降,同时存活细胞的数量显著性减少。与正常糖培养相比较,高糖培养下人肾小球系膜细胞的炎症因子IL-6以及IL-8 mRNA的表达显著性增高。结论高糖环境下miR-302c的表达上调,并靶向下调TIMP3的表达,可能促使炎症因子分泌增加、参与高糖环境下对人肾小球系膜细胞的损伤作用。

Objective To investigate the effect of micro-RNA-302c(miR-302c)on the function of human glomerular mesangial cells in high glucose environment.Methods Dual-luciferase reporting assay was used to verify whether miR-302c was involved in the regulation of TIMP3 expression.Human glomerular mesangial cells were cultured in the high glucose environment,and the relative expression levels of miR-302c and TIMP3 mRNA were detected by real-time quantitative fluorescence PCR(qRT-PCR),the survival number of glomerular mesangial cells was detected,and the mRNA expressions of inflammatory cytokines IL-6 and IL-8 were detected by qRT-PCR.Results The dual luciferase reporter assay confirmed that TIMP3 was the target gene of miR-302c.Compared with the normal glucose group,the miR-302c expression of human glomerular mesangial cells was significantly increased,and the expression of TIMP3 mRNA was significantly decreased in the high glucose group.The number of surviving cells was significantly decreased in the cultured human glomerular mesangial cells under the high glucose concentration compared with the normal glucose concentration.Compared with normal glucose culture,the mRNA expressions of inflammatory cytokines IL-6 and IL-8 in human glomerular mesangial cells with high glucose culture were significantly increased.Conclusion The expression of miR-302c up-regulated in high glucose environment,and it can target to down-regulate the TIMP3 expression,It might promote the secretion of inflammatory factors,and probably involved in the damage of human glomerular mesangial cells under high glucose environment.