The Expression And Significance Of Nestin And Cleaved Caspase-3 In Glomeruli Of Lupus Nephritis

Object: MRL/faslpr mice were used in this study to investigate the expression change of nestin and cleaved caspase-3 in mouse podocyte and its relationship with mice proteinuria level.Methods: Female MRL/MPJ and MRL/faslpr mice were respectively set as Control group and LN group, and urine protein and serum biochemical levels were measured to detect the mice kidney function; The expression of cleaved caspase-3 protein in mice glomeruli were detected by immunohistochemistry and western blot; in addition, the expression of nestin protein in mice glomeruli were detected by immunohistochemistry; And the co-localization level of nestin and cleaved caspase-3 protein were analysis by immunofluorescence.Results:1 The 24 h Upro level of 30-week female MRL/faslpr mice was increased. Compared with Control group, the 24 h Upro level of LN group mice was increased, and the difference was statistically significant; However, there was no significant difference in BUN and Scr level between two groups.2 The podocyte foot fusion was observed in mice glomeruli of LN group. The result of transmission electron microscopy showed that there was no significant structure change of glomeruli in Control group, and increased thickness of glomerular basement membrane and the podocyte foot fusion in the glomeruli were observed in LN group.3 The expression of cleaved caspase-3 protein was increased in glomeruli of LN group. Immunohistochemistry showed that, the positive signal of cleaved caspase-3 protein was shown as yellow granular, and mainly located in the nuclear and(or) cytoplasm of glomeruli cells. In Control group, no obviously positive signal was detected in mice glomeruli, and in LN group, the expression level of cleaved caspase-3 protein was obviously up-regulated. Western blot result showed that, the relative expression level of cleaved caspase-3 in mice glomeruli of LN group was significant increased then that in Control group.4 The expression of nestin was increased in glomeruli of LN group. Immunohistochemistry showed that, the positive signal of nestin protein was shown as yellow granular, and mainly located in the nuclear and cytoplasm of glomeruli cells. Compared with Control group, the expression level of nestin protein in mice glomeruli cells of LN group was significantly increased.5 The expression of cleaved caspase-3 in mice podocyte of LN group was increased. Immunofluorescence showed that, in glomeruli of Control group there was no obviously co-localization of nestin and cleaved caspase-3 protein, and in glomeruli of LN group significant yellow positive co-localization signal was detected.6 Correlation analysisi: In LN group, the 24 Upro level of mice was significantly positive correlated with the expression quantity of cleaved caspase-3 protein in glomeruli cells(r=0.827, P=0.006).Conclusions: the expression levels of cleaved caspase-3 and nestin protein in mice podocyte of LN group were both significantly up-regulated, and there was obviously co-localization of them, the 24 Upro level of mice was significantly positive correlated with the expression quantity of cleaved caspase-3 protein in glomeruli cells. All the results indicate that, the cytoskeleton change and apoptosis of podocyte maybe involved in the pathogensis of LN.