Effects Of Leech On Podacalyxin And VEGF In Podocytes Of Focal Segmental Glomerulosclerosis In Rats

Objective:To investigate the effect of Leech on the serum creatinine and 24 hrUprV of focal segmental glomerular sclerosis,to provide a theoretical basis for leech treatment of focal segmental glomerular sclerosis.Methods: In the experiment,after using arteria renalis resection,the adriamycin nephropathy model was established by injecting adriamycin two 4mg/kg and 3mg/kg into the tail vein.The experimental animals were divided into four groups: blank group,sham operation group,model group and treatment group.The treatment group was treated with leech(0.9g/kg/d),sham-operation group and model group were treated with distilled water,blank group normal feeding.After 4 weeks and the last 8 weeks of treatment,the general state of the experimental rats,the changes of 24 h urine protein and serum creatinine were observed,and pathological changes of renal tissue were observed by PAS and MASSON staining of renal pathological sections;the ultrastructure of kidney was observed by electron microscope;immunofluorescence staining was used to observe the changes of Podacalyxin expression in podocytes;The expression of Podacalyxin and vascular endothelial growth factor(VEGF)protein in renal tissue were detected by immunohistochemical two step method;The expression of Podacalyxin and VEGF in renal tissue was detected by real-time fluorescence quantitative PCR.Results:1.leech can improve the general state of adriamycin nephropathy in rats;in the treatment group,the levels of 24 h urinary protein and serum creatinine were lower than those in the model group,and the difference was statistically significant(P<0.05);2.Leech can relieve the pathological changes of FSGS;3.Immunohistochemical results showed:Model group and treatment group compared with the control group,the reduction of the expression of podocalyxin in glomerular visceral epithelial cells,the differences were statistically significant(P<0.05),the treatment group compared with the model group,the expression of podocalyxin was significantly increased,the difference was statistically significant(P<0.05);model group,treatment group compared with the control group,increased expression of VEGF in glomerular visceral epithelial cells,the differences were statistically significant(P<0.05),the treatment group compared with the model group,the expression of VEGF was significantly decreased,the difference was statistically significant(P<0.05);4.RT-PCRresults show : Model group and treatment group compared with the control group,the expression of podocalyxinm RNA decreased,the differences were statistically significant(P<0.05),the treatment group compared with the model group,the expression of podocalyxin increased,the difference was statistically significant(P<0.05);model group,treatment group compared with the control group,the expression of VEGFm RNA increased,the differences were statistically significant(P<0.05).The treatment group compared with the model group,the expression of VEGFnmRNA was significantly reduced,the difference was statistically significant(P<0.05).Conclusions:1.Leech treatment group can reduce FSGS rats 24 h urine protein quantitative,reduce serum creatinine;2.leech can reduce the pathological changes of kidney and protect the kidney;3.The expression of podocalyxin in renal tissue of rats with adriamycin nephropathy was enhanced and the expression of VEGFmRNA was decreased,which could maintain the structure and function of podocytes,and delay the development of FSGS.