Based On The JAK/STAT/SOCS Signaling Pathway Study The Effect And Mechanism Of Qishishenshu Capsule On Diabetic Nephropathy Rats

Objective: By observing the effect of Qishishenshu capsule on the JAK/STAT/SOCS signaling pathway in diabetic nephropathy rats,the effect and mechanism of Qishishenshu capsule on diabetic nephropathy rats were discussed.Methods:The 50 male rats of clean grade SD were randomly divided into control group,model group,low-dose group of Qishishenshu capsule(Qishishenshu capsule),high-dose group of Qishishenshu capsule(Qish-ishenshu capsule)and valsartan group according to the random number table,with 10 rats in each group.In addition to the control group,rats in other groups were fed a high-fat and high-sugar diet and intraperitoneal injection of streptozotocin(STZ)(35mg/Kg)to induce the establishment of diabetic nephropathy rat model.The control group and the model group were given normal saline gavage,and the other groups were given qishishenshu capsule low-dose(1.2g/kg.d),qishishenshu capsule high-dose(2.4g/kg.d)and valsartan capsule(0.034g/kg?d)gavage once a day for 8 weeks.Observation rat generally,fasting,can not help but water every 2 weeks after 12 hours of weighing and measuring,fasting glucose metabolic cage and collect urine test urine trace albumin/uric creatinine(ACR),every 4 weeks quantitative detection of 24 hours urinary protein,the last to be put to death in the rat to return after the treatment the blood samples of renal function,return kidney under light microscopy to observe morphological changes(HE staining and Masson staining);The expressions of P-JAK2,P-STAT3,SOCS1 and SOCS3 were observed by immunohistochemical staining;The mRNA expressions of JAK2,STAT3,SOCS-1 and SOCS-3 in renal tissues were detected by fluorescence quantitative PCR.The protein contents of P-JAK2,P-STAT3,SOCS-1 and SOCS-3 in renal tissues were determined by Western blot.Results:1.Changes of blood glucose and body weight: compared with the control group,blood glucose was significantly increased in the model group and the treatment group(p<0.01),but there was no statistically significant difference in body weight(p>0.05).Compared with the model group,the blood glucose changes in each treatment group were not statistically significant,while the body weight increased significantly(p<0.01),the difference was statistically significant.Compared with the valsartan group,there was no statistically significant difference in blood glucose between the low kidney group and the high kidney group(p>0.05).Weight gain in the low kidney group was not statistically significant(p>0.05),while weight gain in the high kidney group was statistically significant(p<0.05).Compared with the group with high kidney,the changes of blood glucose in the group with low kidney were not statistically significant(p>0.05),while the body weight was significantly reduced(p<0.05).2.ACR and 24-hour urinary protein quantification: Compared with the control group,there was no statistically significant difference in ACR and 24-hour urinary protein quantification before treatment in the model group and each treatment group(p>0.05),and the difference was significantly increased after treatment(p<0.01).Compared with the model group,ACR and 24-hour urinary protein quantification were significantly reduced in each treatment group,with statistically significant differences(p<0.05).Compared with the valsartan group,the ACR and 24-hour urinary protein quantification were increased in the renal low group,but the differences were not statistically significant(p>0.05),while the ACR and 24-hour urinary protein quantification were decreased in the renal high group(p<0.05).Compared with the group with high kidney,the ACR and 24-hour urinary protein quantification were increased in the group with low kidney,but the difference was not statistically significant(p>0.05).3.Indicators of urea nitrogen(BUN)and blood creatinine(CREA): compared with the control group,BUN and CREA in the model group,the kidney group and the kidney group were all increased,with statistically significant differences(p<0.05).Compared with the model group,the level of BUN in the kidney-low group increased(p<0.01)and the level of CREA decreased(p>0.05),while the level of BUN and CREA in the kidney-high group and the valsartan group decreased significantly,but there was no significant difference in BUN(p>0.05)and the level of CREA decreased(p<0.01).Compared with the valsartan group,the levels of BUN and CREA were significantly higher in the low kidney group(p<0.01)and in the high kidney group(p>0.05).Compared with the kidney high group,both BUN and CREA in the kidney low group were higher,but there was no statistical significance(p>0.05).4.Renal pathological changes: HE and Masson staining showed no obvious atrophy or enlargement of the glomerulus,no thickening of the basement membrane of capillaries and no inflammatory exudation in the control group.In the model group,different degrees of inflammatory cell infiltration were observed in the renal interstitium,mainly lymphocytes and neutrophils,and a small amount of fibrous connective tissue hyperplasia and proliferative fibroblast aggregation were observed in the renal interstitium.In the renal hypoxia group,there was more inflammatory cell infiltration in the renal interstitium,mainly lymphocytes,and more renal tubular epithelial cells were observed to be seriously swollen and deformed in a balloonlike manner.The infiltration of inflammatory cells in the renal stroma and the slight balloon-like changes of renal tubular epithelial cells in the kidney-high group were less severe than those in the kidney-low group,and the lesions in the sartan group were similar to those in the kidney-high group.5.Immunohistochemical staining: in the control group,P-JAK2,P-STAT3,SOCS1 and SOCS3 were slightly expressed in the glomerular mesangial area and the renal tubulointerstitial area.Compared with the control group,the expressions of P-JAK2 and P-STAT3 in the model group and the treatment group were relatively increased,while the expressions of SOCS1 and SOCS3 were relatively decreased,but the differences were not statistically significant(p>0.05).The changes of P-JAK2 in the nephrotic group were not statistically significant(p>0.05),while the expression of P-STAT3 was increased when Compared with the model group,the expression of P-JAK2 and P-STAT3 in each treatment group was relatively reduced,while the expression of SOCS1 was relatively increased,and there was no statistically significant difference in the expression of SOCS3(p>0.05).Compared with the valsartan group,there was no statistically significant difference in the expression of P-JAK2,P-STAT3,SOCS1 and SOCS3 in the low-kidney group(p>0.05),and no statistically significant difference in the expression of P-JAK2 and P-STAT3 in the high-kidney group(p>0.05).Compared with the group with high kidney,the expression of P-JAK2 and P-STAT3 in the group with low kidney increased,while the expression of SOCS1 and SOCS3 decreased,but the difference was not statistically significant(p>0.05).6.The mRNA expressions of JAK2,STAT3,SOCS-1 and SOCS-3 in kidney tissues were detected by fluorescence quantitative PCR: compared with the control group,the mRNA expressions of JAK2 and STAT3 in other groups increased,and the mRNA expressions of SOCS1 and SOCS3 decreased,with statistically significant differences(p<0.01).Compared with the model group,the mRNA expressions of JAK2 and STAT3 in each treatment group were decreased,but the difference was not statistically significant(p>0.05),and the mRNA expressions of SOCS1 and SOCS3 were significantly increased(p<0.01).Compared with each treatment group,JAK2 and STAT3 mRNA expression was the highest in the renal low group,while SOCS1 and SOCS3 mRNA expression was the lowest in the renal high group,while JAK2 and STAT3 mRNA expression was the lowest in the renal high group,while SOCS1 and SOCS3 mRNA expression was the highest in the valsartan group,but the difference was not statistically significant(p>0.05).7.Western Blot detection of the contents of P-JAK2 and P-STAT3 as well as SOCS-1 and SOCS-3 in renal tissues: compared with the control group,the expressions of P-JAK2 and P-STAT3 in other groups were significantly increased,while the expressions of SOCS1 and SOCS3 were significantly decreased(p<0.01).Compared with the model group,the expressions of P-JAK2 and P-STAT3 in each treatment group were significantly decreased(p<0.01),while the expressions of SOCS1 and SOCS3 were significantly increased(p<0.01).Compared with the valsartan group,the expressions of P-JAK2 and P-STAT3 were increased and the expressions of SOCS1 and SOCS3 were decreased in the low-kidney group,and the expressions of P-JAK2 and P-STAT3 were decreased in the high-kidney group,while the expressions of SOCS1 and SOCS3 were increased in the high-kidney group,with statistically significant differences(p<0.05).The expression of P-JAK2 and P-STAT3 increased in the low-kidney group,while the expression of SOCS1 and SOCS3 decreased in the low-kidney group(p<0.01).Conclusion:1.Qishishenshu capsule can reduce urinary microalbumin/urinary creatinine ratio and 24-hour urinary protein ration in diabetic nephropathy rats.2.Activation of the JAK/STAT/SOCS signaling pathway is involved in the development of kidney damage in diabetic nephropathy rats.3.The protective effect of Qishenshu capsule on the kidney of diabetic nephropathy rats may be related to down-regulation of the expression of P-JAK2 and P-STAT3 in the JAK/STAT/SOCS signaling pathway and up-regulation of the expression of SOCS1 and SOCS3.