| Part One:Clinical study Objective:To make clear the effect of Shengqing Jiangzhuo Capsule on intestinal barrier function and micro inflammatory state of CKD patients through clinical study.Method:The patients were divided into the treatment group and the control group by a randomized controlled trial,with CKD3-5(non dialysis)patients as the research subjects.The control group was treated with routine treatment.On the basis of routine treatment,the treatment group was given Shengqing Jiangzhuo Capsule 5 capsules,3 times per day,and the course was 8 weeks.The patient's age and sex,primary disease,clinical symptoms were collected.And the blood,urine test were checked,stool samples were collected both before and after treatment.(1)The baseline clinical data,TCM symptom scores,renal function and micro inflammatory indexes of two groups were compared,so as to confirm the effects of Shengqing Jiangzhuo Capsule on clinical symptoms,renal function and micro inflammatory in CKD patients.(2)To compare the intestinal physical barrier index IS,LPS,immune barrier index fecal sIgA between the two groups,to clarify the effect of Shengqing Jiangzhuo Capsule on intestinal barrier function of CKD patients.Results:A total of 60 patients were enrolled.Finally,52 patients completed the trial,24 in the control group and 28 in the treatment group.(1)The effect of Shengqing Jiangzhuo Capsule on the symptoms of Chinese medicine of CKD patients:The symptom score of TCM syndromes in two groups of patients before and after treating was compared.The results showed that after treating in the treatment group,the scores of it was significantly lower than before treating and the control group after treating(P<0.05).The differential value before and after treating of the treatment group was significantly higher than control group(P<0.05).There was no significant difference in the score of the control group before and after treating(P>0.05).The effect of Shengqing Jiangzhuo Capsule on gastrointestinal symptoms was further analyzed.After treatment,the anorexia,nausea and vomiting,abdominal distension symptom of treatment group were statistically significant compared with the control group(P<0.05),and the anorexia,abdominal distension symptom were statistically significant compared with those before treating(P<0.05).The severity of these symptoms after treatment in the treatment group was significantly lower than that in the control group and before treatment.(2)The effect of Shengqing Jiangzhuo Capsule on the safety index of CKD patients:After treating,there was no significant difference in ALT,AST and K between the control group and the treatment group(P>0.05),and there was no significant difference in the above indexes between the treatment group and the control group after treating(P>0.05).(3)The effect of Shengqing Jiangzhuo Capsule on Blood biochemical indexs of CKD patients:After treating,there was no significant difference in Hb?ALB?TG?TC?Ca?P?PTH?TCO2 between the control group and the treatment group(P>0.05),and there was no significant difference in the above indexes between the treatment group and the control group after treating(P>0.05).While the UA in the treatment group after treating was significantly lower than it in the control group(P<0.05),and decreased significantly compared with that before treating(P<0.05).(4)The effect of Shengqing Jiangzhuo Capsule on renal function of CKD patients:After treating,the ACR in the treatment group was significantly lower than those in the control group(P<0.05).Although there was no significant difference in eGFR between the treatment group and the control group after treating(P>0.05),the difference value of eGFR between the treatment group and the control group was significantly larger than that of the control group(P<0.05).(5)The effect of Shengqing Jiangzhuo Capsule on the micro inflammation of CKD patients:The IL-6 and TNF-? in the treatment group after treating were significantly lower than those before treating treating,and significantly lower than those in the control group after treating(P<0.05).However,there was no significant difference between the two groups before and after treating in CRP(P>0.05).(6)The effect of Shengqing Jiangzhuo Capsule on the intestinal barrier function of CKD patients:The IS and LPS in the treatment group after treating were significantly lower than those before treating(P<0.05),and significantly lower than those in the control group after treating(P<0.05).There was no significant difference between the two groups of fecal sIgA before and after treating(P>0.05).The correlation between sIgA and eGFR before treating was analyzed,and there was a positive correlation between the two indexes.Comparing the amount of fecal sIgA before treating with different CKD stage,it was found that the difference among the three groups was statistically significant(P<0.05),and the amount of fecal sIgA in CKD5 stage was the least.The amount of fecal sIgA in the two groups was compared at different CKD stages.The fecal sIgA in the CKD5 stage treatment group after treating was significantly higher than that before treating(P<0.05),and was significantly higher than that in the control group after treating(P<0.05).Conclusion:It is clear that Shengqing Jiangzhuo Capsule has protective effect on intestinal barrier function and alleviating micro inflammatory reaction in CKD patients.And it is found that it has the effect of lowering serum uric acid,so as to provide clinical evidence for further promoting its use.It is also found that the amount of fecal sIgA is becoming less and less with the progress of CKD.Part Two:Experimental study Objective:To elucidate the mechanism of Shengqing Jiangzhuo Capsule in treating chronic kidney disease through experimental study.To further explore the effect target of Shengqing Jiangzhuo Capsule Based on the theory of Gut-kidney Axis and intestinal TLRs/NF-? B signal transduction pathway so as to provide the basis for the treatment of CKD with Shengqing Jiangzhuo Capsule.Method:5/6 nephrectomy model was used to establish the CRF model in rats.Set up a normal group,a sham operation group,a model group and a Shengqing Jiangzhuo Capsule treatment group.5 rats in each group were tested.The intervention began second days after the success of the model,and the total intervention was 8 weeks.After the intervention,the blood was collected,the urine were preserved for 24 hours,faeces were collected,and the kidneys,ileum and colonic tissues were retained.(1)The serum creatinine,urea and 24 hours urine protein were compared in different groups.Pathological changes of renal tissue in rats were compared,and the degree of renal fibrosis in different groups was compared.(2)The serum IL-6,TNF-?,IS,LPS and fecal sIgA that detected by ELISA method were compared in different groups.And the correlation of each index was analyzed.(3)Light microscopic of intestinal mucosa in rats were checked,to compare the intestinal mucosal pathology changes of different groups.Compare the expression of tight junction protein claudin-1,occludin and ZO-1 in the intestinal mucosa of different groups by Western-blot method.Using Real-time PCR technique to detect TLR2,TLR4,and TLRs/NF-? B signal transduction pathway's key protein MyD88,NF-? B mRNA expression in intestinal tissue,compare the mRNA expression of aboves of different groups.Analyze the relationship between tight junction protein and the micro inflammation index.Results:(1)The effect of Shengqing Jiangzhuo Capsule on renal function of 5/6 nephrectomy rats:In the indexes of renal function,the serum creatinine,urea of model group and treatment group were significantly higher than normal group(P<0.05).There was no significant difference between normal group and sham operation group(P>0.05).The serum creatinine,urea of treatment group was significantly lower than model group(P<0.05).In the 24 hour urine protein quantitative,model group and treatment group were significantly higher than normal group(P<0.05),while treatment group was significantly lower than model group(P<0.05).In renal pathology,normal group and sham operation group show the normal renal tissue in HE staining,while model group showed obvious fibrosis pathological changes.And treatment group was significantly improved compared with the model group.In masson staining,the semi quantitative analysis results showed that model group and treatment group were significantly higher than the normal group and sham operation group in kidney fibrosis level(P<0.05).Treatment group was significantly lower than model group in kidney fibrosis level(P<0.05).Treatment group decreased by about 70.5% compared with model group in fibrosis score.(2)The effect of Shengqing Jiangzhuo Capsule on the micro inflammation of 5/6 nephrectomy rats:The levels of IL-6 and TNF-? in model group and sham operation group were significantly higher than normal group(P<0.05).Treatment group and sham operation group were significantly lower than model group(P<0.05).While there was no statistical significance between treatment group and normal group(P>0.05).The levels of IS and LPS in model group were significantly higher than those in normal group and sham operation group(P<0.05).The levels of IS and LPS in treatment group were significantly lower than those in model group(P<0.05).There was no significant difference among sham operation group,treatment group and the normal group(P>0.05).There was no significant difference in the fecal sIgA among normal group,sham operation group,model group and treatment group(P>0.05).Pearson correlation analysis showed that there is a positive correlation between LPS and IL-6,also LPS and TNF-?.And there is no significant correlation between LPS and fecal sIgA.IS and IL-6,IS and TNF-? were both positively related.(3)The effect of Shengqing Jiangzhuo Capsule on the intestinal barrier of 5/6 nephrectomy rats:Intestinal pathology HE staining showed that the intestinal villi of model group were smaller and shorter than those of normal group and sham operation group,and the gland proliferation of model group was not active.The colonic mucosa edema and the inflammation of the lamina propria were serious in model group.Treatment group was obvious improved compared to model group.The expression of tight junction protein in intestinal tissue was detected by Western-blot method.The expression of the three TJPs in model group was lower than those of the normal group and the sham operation group(P<0.05).The expression of the three TJPs in treatment group increased significantly higher than those in model group(P<0.05).Using Real-time PCR technique to detect TLR2,TLR4,MyD88,NF-? B mRNA expression in intestinal tissue.The expression of TLR2 mRNA in model group was significantly lower than that in normal group,sham operation group and treatment group(P<0.05).There was no statistical significance between treatment groups and normal group in TLR2 mRNA expression(P>0.05).There was no significant difference in TLR4 mRNA expression between model group and normal group(P>0.05).The TLR4 mRNA expression in sham operation group and treatment group was significantly higher than that in model group(P<0.05).The expression of MyD88 and NF-? B mRNA in model group and sham operation group were significantly higher than those in normal group(P<0.05).The expression of MyD88 and NF-? B mRNA in treatment group were significantly lower than those in model group(P<0.05).One of the representative of TJPs ZO-1 and intestinal toxins IS,TLR2 mRNA expression,LPS and TLR4 mRNA expression were analyzed by Pearson correlation analysis.the results showed that ZO-1 and IS,ZO-1 and LPS were both negatively correlated,and positively correlated between ZO-1 and TLR2 mRNA expression.There is no significant correlation between ZO-1 and TLR4 mRNA expression.The correlation between TLR4 mRNA expression and IL-6,TNF-? was analyzed.And there is no significant correlation was found(P>0.05).The correlation analysis between NF-? B mRNA expression and IL-6,TNF-?,ZO-1 showed that NF-? B mRNA expression was positively correlated with IL-6 and TNF-?,and negatively correlated with the ZO-1.Conclusion:It was proved that Shengqing Jiangzhuo Capsule can delay the 5/6 nephrectomy rat renal interstitial fibrosis,reduce micro inflammation,reduce intestinal toxins LPS and IS in the circulation,repair of intestinal barrier function.Finally,the hypothesis was proved that Shengqing Jiangzhuo Capsule delay renal interstitial fibrosis by regulating the TLRs/NF-? B signal transduction pathway.The deeper mechanism of Shengqing Jiangzhuo Capsule in treating CKD was elucidated.And it was found that the effect was multi target and multi-level. |
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