Effect of miR-92b-5p on podocyte apoptosis induced by hyperglycemia and its mechanism
Objective To investigate the possible role of miR-92b-5p in podocyte injury or apoptosis induced by hyperglycemia,and explore its effect on mitochondrial function and its molecular mechanism.Methods ① The podocytes were cultured and divided into normal glucose(NG)group and high glucose(HG)group,and the differentially expressed miRNAs were screened using high-throughput RNA-sequencing(RNA-seq)technology.②In order to investigate the effect of miR-92b-5p on podocyte mitochondrial function and podocyte apoptosis,the podocytes under normal glucose condition were divided into NC mimics group,miR-92b-5p mimics group,NC inhibitor,and miR-92b-5p inhibitor group.In order to investigate the effect of miR-92b-5p on podocyte apoptosis and mitochondrial function,the podocytes were divided into NG group,HG group,HG+NC inhibitor group,and HG+miR-92b-5p inhibitor group.Real-time polymerase chain reaction(qRT-PCR)was used to determine the expression of miR-92b-5p,flow cytometry was used to measure the podocyte apoptosis rate,CCK-8 was used to measure the podocyte viability,and Western blot was used to measure the expressions of apoptosis-related proteins and mitochondrial-related proteins.Results ①Through high-throughput sequencing technology,16 differentially expressed miRNAs were identified,among which the expression of miR-92b-5p in HG group was significantly increased(P＜0.05).②Compared with NC mimics group,the expression of miR-92b-5p was increased in miR-92b-5p mimics group(P＜0.05),the cell viability was decreased(P＜0.05),the podocyte apoptosis rate was increased(P＜0.05),the expressions of Bcl-2 and Mfn1 were decreased(P＜0.05),and the expressions of Bax and Fis1 were increased(P＜0.05).③Compared with NG group,the expression of miR-92b-5p was increased in HG group(P＜0.05),the podocyte viability was decreased(P＜0.05),the podocyte apoptosis rate was increased(P＜0.05),the expressions of Bcl-2 and Mfn1 were decreased(P＜0.05),and the expressions of Bax and Fis1 were increased(P＜0.05).Compared with HG+NC inhibitor group,the expression of miR-92b-5p was decreased in HG+miR-92b-5p inhibitor group(P＜0.05),the cell viability was increased(P＜0.05),the podocyte apoptosis rate was reduced(P＜0.05),the expressions of Bcl-2 and Mfn1 were increased(P＜0.05),and the expressions of Bax and Fis1 were decreased(P＜0.05).Conclusion Under high glucose condition,the expression of miR-92b-5p increases.Inhibition of miR-92b-5p could reduce the podocyte apoptosis induced by hyperglycemia by improving mitochondrial function,thus delaying the progression of diabetic nephropathy.
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