The Role Of IQGAP1 In The Regulation Of Cytoskeletal Rearrangement In Podocytes

Objective:The experimental study of mouse podocytes in vitro is carried out by using molecular biology method,To investigate the regulatory role of IQGAP1 in podocytes cytoskeletal rearrangement.Methods:The animal model of minutely denatured nephropathy induced by purinamycin has a certain similarity in clinical manifestaition and pathological changes,the most characteristic pathological change of small denatured nephrosis is the extensive fusion of the foot process,the reorganization of the cytoskeleton protein of the podocyte plays a key role in the fusion of podocyte process.Conditionally immortalized mouse podocytes were used to perform cell culture and stimulated by puromycin aminonucleoside(PAN).RT-PCR and Western blot analysis were used to measure the level of IQGAP1 mRNA and protein-respectively.The cytoskeleton of F-actin was evaluated by FITC-conjugated phalloidin staining under laser confoc al scanning microscope and semi-quantitative system with cortical F-actin score(CF S)was used to analyze the degree of actin cytoskeleton arrangement.Then we further explored the changes of cytoskeletal rearrangement after up-and down-regulation of IQGAP1 using plasmid transfection technique and gene silencing technique.SPSS22.5 software was used for statistical analysis.Measurement data are represented by mean number± standard deviation(x±s),the parameter data were analyzed by single factorANOVA and LSD was compared between groups.The correlation analysis indicated the correlation between the two variables.The difference was statistically significant with P<0.05.Results:Repeated three tests were carried out smoothly(1)when compared with control group,the expression ofIQGAP1 mRNA and protein was significantly decreased after PAN stimulation in mouse podocytes(P<0.05).PAN stimulation induced.cytoskeletal rearrangement including cortical F-actin ring formation,stress fiber attenuation and CFS up-regulation;(2)the above rearrangement was exacerbated bylQGAP1 siRNA transfection(P<0.05);(3)PAN induced cortical F-actin ring formation and CFS up-regulation was abated after IQGAP1 plasmid transfection(P<0.05).Conclusion:IQGAP1caninhibit PAN induced cytoskeleton rearrangement in podocytes and plays an important role in maintaining normal podocyte cytoskeleton.