Uremic Clearance Granule Alleviates Renal Dysfunction And Renal Fibrosis In 5/6nephrectomized Rats By Upregulating Klotho Expression

Objective:The prevalence and mortality of chronic kidney disease(CKD)has increased year by year and has become one of the most important public health issues of global concern.Klotho is a gene closely related to the occurrence and progression of CKD.Preventing the decline of Klotho and re-activating endogenous Klotho to produce or supplement exogenous Klotho can attenuate renal fibrosis and delay the progression of CKD.It can be used as a potential therapeutic target for CKD.Uremic clear granules(UCG)is an effective traditional Chinese medicine preparation for the treatment of CKD.It has the characteristics of multi-component,multi-target and multi-channel synergy.The pharmacological mechanism is complex,and relevant in-depth research is still lacking.In this study,the rat model of CKD was prepared by 5/6 nephrectomy.The effect of uremic clearing on renal function and renal fibrosis in CKD rats and its mechanism of action were investigated.It is explored whether UCG may play a pharmacological role by regulating the expression of endogenous Klotho.Method:Thirty-six SD rats were randomly divided into three groups:sham operation group(Sham group,sham operation);model group(Model group,5/6 nephrectomy+high-phosphorus diet);uremic acid group(UCG group,5/6)Renal resection +high-phosphorus diet group+uremic clear granules;rat CKD model was established by 5/6 nephrectomy,blood was collected at 2 weeks,4 weeks,8 weeks,and 12 weeks after operation,and serum biochemical indexes of each group were detected.:creatinine(SCr),urea nitrogen(BUN);Renal tissue was taken 12 weeks after the modeling operation,and some were used for Western and RT-PCR to detect the expression of Klotho,TGF-?1,a-SMA,FN.The other part is used for HE staining,Masson staining and immunohistochemistry.Result:General situation:After the operation,the model rats gradually became yellow and messy and easy to remove hair,and the spirit was wilting.The above performance was lighter in the UCG group.The residual kidney weight and kidney index of the UCG group were significantly lower than the model group at 12 weeks after surgery(P<0.05).Appearance of the kidney:The kidney of the normal group was rosy and smooth.The residual kidney of the model group was hypertrophied,the surface was pale and ischemic,the renal pelvis was highly dilated(with urine accumulation),and the UCG group was ameliorated.Serum biochemical parameters:BUN and Scr were significantly increased in the 5/6 nephrectomied rats at 2 weeks after operation,and BUN and Scr in the model group continued to increase.UCG intervention can maintain renal function in rats.After 10 weeks of administration,The BUN and Scr level in UCG group was significantly lower than the model group(P<0.001).Renal tissue HE staining and Masson staining:there is different degrees of damage to the kidney tissue of the model rats,and the renal pathological damage in the UCG group was improved compared with the model group.Quantitative analysis of pathological images:the histopathological scores of the rats in the model group were significantly higher than those in the sham operation(P<0.001).The pathological damage of the glomeruli and renal tubules in the UCG group were lower than those in the model group(P<0.05).Western Blot and RT-PCR results showed that the expression of Klotho in UCG group was higher than that in model group(P<0.05).The expression of TGF-?1,?-SMA and FN in UCG group was lower than that in model group(P<0.05).The results of immunohistochemistry showed that the expression of Klotho in the renal tissue of the model group was decreased,and the expression of Klotho was up-regulated by UCG.Conclusion:UCG can improve renal function in 5/6 nephrectomized rats,reduce renal fibrosis and delay the progression of CKD.UCG may down-regulate the expression of renal injury molecule TGF-?1 by up-regulating the expression of protective molecule Klotho,thus EMT-related molecule E-cadherin expression is up-regulated,a-SMA and FN expression are down-regulated,and renal tubular epithelial transdifferentiation is inhibited.