The Role Of Wnt/β-catenin Pathway And MiR-200a In The Process Of Human Podocyte Injury Induced By High Glucose

ObjectiveTo investigate expression of β-catenin, podocyte injury marker nephrin and miR-200a in HPC cultured with normal glucose or high glucose.To detect expression of miR-200a, β-catenin and nephrin in HPC cultured with normal glucose or high glucose which has been transfected with miR-200a-3p mimics previously. To investigate whether miR-200a-3p mimics can inhibit Wnt/β-catenin transduction and attenuate injury of HPC stimulated by high glucose.MethodsDifferentiated human podocyte randomed into5groups:normal control group,25mM high glucose treatment HPCs6hrs,12hrs,24hrs and25mM MN intervene HPCs24hrs group. The expressions of β-catenin, nephrin and miR-200a were examined by realtime PCR, β-catenin, nephrin were also examined by Western blot.Differentiated human podocyte transfected with miR-200a-3p mimics randomed into5groups were exposed to25mM concentrations of high glucose with different duration. The expressions of miR-200a, β-catenin and nephrin were examined by realtime PCR, β-catenin, nephrin were also examined by Western blot.Results(1) There are expression of nephrin and miR-200a in HPC cultured with normal glucose and a little expression of β-catenin.(2)Human podocyte stimulated by high glucose upregulated expression of β-catenin mRNA and protein. Level of p-catenin mRNA in MN group has not statistically significant versus that in control group.(3)Human podocyte stimulated by high glucose downregulated expression of nephrin mRNA and protein. Level of nephrin mRNA in MN group had not statistically significant versus that in control group.(4) Expression of miR-200a was gradually downregulated of human podocyte stimulated by high glucose. miR-200a level had statistically significanin in the HG12h and HG24h group versus that in control group.(5) Expression of miR-200a was increased significantly in HPC after transfection.(6)Human podocyte transfected with miR-200a-3p mimics inhibited expression of p-catenin.(7)Human podocyte transfected with miR-200a-3p mimics upregulated expression of nephrin mRNA and protein and attenuated the injury process induced by high glucose.Conclusion (1)Human podocyte stimulated by high glucose upregulated expression of β-catenin versus downregulated expression of nephrin.(2) Expression of miR-200a in HPC is existed and the level was gradually downregulated in HPC stimulated by high glucose.(3) Wnt/β-catenin pathway and miR-200a possibly may involve in the process of human podocyte injury induced by high glucose.(4)Transfecting with miR-200a-3p mimics into human podocyte not only inhibited the Wnt/β-catenin signaling tranduction, but also upregulated the expression of nephrin and attenuated the injury process induced by high glucose. The thesis comprises12figures and25references.