Effects Of Shendi Granules On PBMC Apoptosis?MCP-1 And TGF-?1 In Chronic Nephritis And Mesangial Prolife Rative Nephritis Rats

Objective To observe the effect of Shendi granules on PBMC apoptosis and serum MCP-1?TGF-?1 in patients with chronic nephritis with deficiency of spleen and kidney.To investigate the therapeutic effect of Shendi granules on chronic nephritis in both cellular and humoral immunity.Methods After the clinical trial was approved by the Ethics Committee of the first affiliated Hospital of Anhui University of traditional Chinese Medicine.60 patients who met the inclusion criteria of chronic nephritis from March 2017 to December 2017 were collected.They were randomly divided into two groups for 12 weeks treatment: treatment group(Shendi granules,10g/time)and control group(valsartan,80mg/d).Observe the changes of indicators before and after drug intervention in the two groups,including renal function(Scr,BUN,e GFR),24 H urinary protein quantitation(24HUPr),PBMC apoptosis,MCP-1,and TGF-?1 changes.Another 20 healthy people in our hospital were selected as normal group.Their blood samples were collected to detect the expression of MCP-1?TGF-?1 and PBMC apoptosis rate.Results1.During the treatment,one case of the treatment group was treated with corticosteroids in the external hospital,while 2 cases in the control group were treated with Tripterygium Glycosides tablets in the external hospital.The final treatment group was 29 cases and the control group was28 cases.A total of 57 cases completed the experiment.2.The results of independent sample t-test showed that there was no statistical difference(P>0.05)in the TCM syndrome scores before treatment between the two groups,but decreased after treatment.T test results of paired samples showed that there was statistical difference in TCM syndromes score before and after treatment(P<0.05).After treatment,the treatment group was superior to the control group(P<0.05).3.There was statistical difference in improving the curative effect of TCM syndrome between the two groups(P<0.05).The treatment group was superior to the control group(P<0.05).4.The judgement of clinical disease curative effect: The difference of clinical effect after treatment in the two groups was statistically significant(P<0.05).The treatment group was better than the control group.5.24 HUPr comparison: Repeated measurement multivariate analysis of variance showed that there was no significant difference in 24-HUPr between visit 1(0 week)and visit 2(4 weeks)in both groups(P>0.05).The decrease of 24 HUPr in the treatment group was better than that in the control group(P<0.05).The curative effect of the treatment group was positively correlated with the time of treatment.6.Comparison of PBMC apoptosis and its apoptosis-regulating protein Bcl-2 and Fas expression:Compared with the normal group,the apoptosis rate of PBMC in the two groups before treatment was significantly higher than that in the normal group(P<0.01).The expression of Bcl-2 decreased and the expression of Fas increased in PBMC before treatment in both groups(P<0.05).Compared with before treatment,the apoptosis rate of PBMC in the control and treatment groups decreased after treatment(P<0.05).After treatment,the expression of Bcl-2was up-regulated and Fas expression was down-regulated in PBMC after treatment(P<0.05).The treatment group was better than the control group in improving PBMC apoptosis rate(P<0.05).7.Comparison of serum MCP-1 and TGF-?1 expression:Before treatment,the expression of serum MCP-1 and TGF-?1 was up-regulated in the two groups compared with the normal group(P<0.05).The expression was down-regulated after treatment(P<0.05).The treatment group was superior to the control group(P<0.05).8.There was no significant change in renal function index(Scr?BUN?e GFR)between the two groups before and after treatment(P>0.05).9.Comparison of safety indexes: there was no significant change in the safety index before and after the clinical study in the treatment group.And no adverse drug reaction was found in the course of treatment.Conclusion1.Shendi granules can significantly improve the clinical symptoms of CGN spleen and kidney deficiency syndrome patients and reduce urinary protein.2.Shendi granules can down-regulate the expression of Fas and up-regulating Bcl-2 expression of PBMC apoptosis protein in order to decrease the rate of PBMC apoptosis.Thereby reducing the immune inflammation of the kidney.The pathological damage of kidney was alleviated.3.Shendi granule can also down-regulate the expression of MCP-1 and TGF-?1 and alleviate the inflammation of glomeruli.4.No adverse reaction to the patient's administration of Shendi granules was found during the treatment.Observed before and after treatment,no significant changes in safety indicators.It shows high security.Objective The rat model of mesangial proliferative glomerulonephritis(MsPGN)was established to observe the apoptosis and apoptotic regulation protein bcl-2 and Fas expression levels in rats.And the expression of inflammatory cytokines MCP-1 and TGF-?1 was observed.In addition,24 HUPr and renal pathological conditions of rats were observed to investigate the intervention effects of Shendi granules on immune inflammatory disorders in MsPGN rats.Methods The male SD rats in the cleaning stage weighed about 200 g.All rats were in the experimental animal room of Anhui University of traditional Chinese medicine for 1 weeks.Ten rats were randomly selected as normal group and MsPGN models was produced in the remaining rats.Then randomly divided into experimental group,control group and model group.The rats were given the drug on the 21 st day after the models was completed.The control group was given valsartan 2ml(dose: 1.03mg/100g·d)daily.The treatment group was treated with 2ml(dose of0.8g/200g·d).The model group and normal group were given physiological saline 2ml.The three groups lasted 12 weeks.24 hours of urine was collected at 4 weeks before and after group entry.The rats were anesthetized by volume at the end of the course.Then the celiac artery and the isolated kidney tissue were used for the following test.First,Scr,BUN,endogenous creatinine clearance rate and 24 HUPr were detected.Second,the expression of serum and kidney MCP-1and TGF-?1 were detected.Thirdly,PBMC apoptosis rate and expression level of bcl-2 and Fas were detected.Finally,the pathological changes of renal tissue were observed under a microscope.Result1.Improve the quality of life of rats: The rats in the normal group had good mental state and activity,and their weight gradually increased.However,the other three groups showed listless lethargy,poor mental status and low diet.The weight was lighter than the normal group.The overall state of the rats after the intervention of the granules and valsartan was improved,which was close to the normal group rats.2.Reduce urinary protein:24HUPr increased in rats after mold making.24 HUPr was decreased after treatment with valsaran and Shendi granules.There was no significant difference of24 HUPr between Shendi granules group and valsartan group in the fourth week of the intervention treatment(P>0.05).Compared with valsartan group,the decrease of 24HUPr(P<0.05)was more obvious in the treatment of the 8 weeks and 12 weeks.This indicates that the Shendi granules has the effect of reducing the 24 H urinary protein in MsPGN rats.Moreover,the therapeutic effect of the treatment time was better than that of valsartan.This is consistent with the study of early Shendi granules.3.Effect on renal function: There was no significant change in BUN,Scr and Ccr before and after treatment in each group(P >0.05).4.Effect of regulating PBMC apoptosis and apoptosis protein:The apoptosis rate of PBMC was significantly increased in the rats after the modeling.This may be related to the immune disorders of MsPGN rats model.After 12 weeks of intervention with granules and valsartan intervention.The apoptosis rate of PBMC in the two groups was lower than that in the model group(P<0.05).The expression of up-regulated bcl-2 and down-regulated Fas is related.And the granules group was better than valsartan group(P<0.05).5.Effect on serum MCP-1and TGF-?1: The expression of MCP-1 and TGF-?1 in rat serum was higher than that in the control group.After 12 weeks of intervention with Shendi granules and valsartan,the expression of MCP-1,TGF-?1 in serum of rats in each group was higher than that in normal group.The expression of MCP-1,TGF-?1 in both groups was decreased(P<0.05),which indicated that both of them could decrease the expression of inflammatory cytokines in MsPGN rat models.However,there was no significant difference in the expression of MCP-1between Shendi granules group and valsartan group(P>0.05).The expression of serum TGF-?1was better than that of valsartan group(P<0.05).6.Effect on renal MCP-1 and TGF-?1:The relative expression levels of MCP-1 and TGF-?1 in kidney tissues were higher than that in normal group.The relative expression levels of MCP-1 and TGF-?1 were decreased in both groups of rats after 12 weeks of intervention with the granules and valsartan intervention(P<0.05).It is suggested that both can improve the renal inflammatory response in MsPGN rat model.Thereby reducing glomerular damage.However,the expression of renal MCP-1 in valsartan group was superior to that of the granules group(P<0.05).There was no significant difference in the expression of TGF-?1 in the kidney.7.The proliferation of mesenchymal mesangial cells and the growth of mesangial matrix were improved in the granule group and valsartan group.And the granule group was superior to valsartan group.Conclusion1.The immune inflammation disorder in MsPGN rats resulted in the increase of apoptosis rate of PBMC,which may be related to the disturbance of the expression of apoptosis-regulating protein Bcl-2 and Fas.2.Shendi granules can improve the survival quality and reduce urinary protein of MsPGN rats.3.Shendi granules may decrease the apoptosis rate of PBMC by up-regulating the expression of Bcl-2 and down-regulating the expression of Fas.4.Shendi granules could reduce the expression of MCP-1 ? TGF-?1 and alleviate the inflammatory reaction of kidney.5.The granules can reduce the proliferation of mesangial cells and mesangial matrix in MsPGN.