Study On The Mechanism Of Action Of Sijunzi Decoction On Intestinal Inflammation And Intestinal Barrier In CKD Mic

Purpose:In the related research of Sijunzi Decoction on the prevention and treatment of CKD,we explored the effects of its mechanism on intestinal inflammation,UTs,intestinal mucosal barrier and kidney structure and function from the intestinal and renal axis theory as the breakthrough point.Therefore,we constructed a 5/6 nephrectomy-induced CKD mouse model,and intervened with Sijunzi Decoction to achieve the effects of improving intestinal inflammation,reducing UTs level,repairing intestinal mucosa,protecting renal function,and delaying renal damage.To provide new treatment methods and ideas for the prevention and treatment of CKD with traditional Chinese medicine.Material and method:In this study,60 male mice with SPF-grade C57BL/6 were divided into 6 groups using a 5/6 nephrectomy model.Sham operation(Sham)group,Model(Model)group,Meichang ’an(MCA)group,Sijunzi Decoction low-dose(SJZT-L)group,Sijunzi decoction medium-dose(SJZT-M)group,Sijunzi decoction high-dose(SJZT-H)group,the Sham group and model group were respectively given the same dose of normal saline.MCA group was given corresponding dose intragastric administration,and SJZT group was given corresponding dose intragastric administration,once a day for 8 weeks.The kidney and colon specimens were collected,blood was collected,serum Scr and BUN were measured,and HE stained the kidney and colon tissues,ELISA method to detect serum IL-6,IS,TMAO,RT-q PCR to detect Occludin,Claudin-1,IL-6m RNA,Western-blot method to detect colon tissue Occludin,Claudin-1 protein content,immunofluorescence method to detect Occludin,colon tissue,Claudin-1 protein content.The experimental data were analyzed statistically by SPSS 23.0、Image J and Graphpad prism 8.0.The experimental results is expressed as the average standard deviation;One-way analysis of variance was used to compare data differences between groups,and P <0.05 indicated that there were statistical differences.Results:1.Serum results show:Compared with the Sham group,serum Scr and BUN levels were significantly higher in the Model group(P <0.05);compared with the Model group,serum Scr and BUN levels were significantly lower in the SJZT-L,SJZT-M,SJZT-H and MCA groups(P <0.05).2.Renal HE staining results showed that:In the Sham group,the glomeruli were intact,no atrophy was seen,and the tubular lumen was normal.compared with the Sham group,the tubular lumen of the Model group was significantly enlarged,glomerular atrophy was evident,the walls of the small arteries were thickened,inflammatory cells were infiltrated,and cell shedding was seen.these pathological histological changes were improved in the SJZT-L,SJZT-M,SJZT-H,and MCA groups.3.HE staining of colon showed that:In the Sham group,the intestinal tissue was structurally intact at all levels,with neatly arranged intestinal villi and abundant intestinal glands,and no significant inflammatory cell infiltration was seen.Compared with the Sham group,the intestinal glands in the Model group were disrupted,the gap was widened,and local inflammatory cell infiltration was seen.Compared with the Model group,the SJZT-L,SJZT-M,SJZT-H and MCA groups showed less intestinal gland destruction,narrower interstitial space and less inflammatory cell infiltration in the lamina propria,with more obvious improvement in the SJZT-H and MCA groups.4.The results of ELISA showed that Compared with the Sham group,serum IL-6,IS and TMAO levels were increased in the Model group(P < 0.05);compared with the Model group,IL-6,IS and TMAO levels were decreased in the SJZT-L,SJZT-M,SJZT-H and MCA groups(P < 0.05),with significant decreases in the SJZT-H and MCA groups compared with the Model group.5.RT-q PCR results showed that Compared with Sham group,the level of IL-6m RNA in colon tissue in Model group increased significantly,while the m RNA of Occludin and Claudin-1 decreased significantly.Compared with the Model group,the levels of IL-6m RNA in SJZT-L,SJZT-M,SJZT-H and MCA groups decreased significantly,while the levels of Occludin and Claudin-1m RNA increased.The above differences were statistically significant(P < 0.05),especially in SJZT-H and MCA groups.6.Western-blot results showed that Compared with the Sham group,the expression of Occludin and Claudin-1 protein in colon tissue decreased in the Model group(P < 0.05);compared with the Model group,the expression levels of Occludin and Claudin-1 protein increased in the SJZT-L,SJZT-M,SJZT-H and MCA groups(P < 0.05),with SJZT-H and MCA groups increased significantly compared with the Model group.7.Immunofluorescence staining showed that Occludin and Claudin-1 protein expression was decreased in the Model group compared with the Sham group(P < 0.05).Compared with the Model group,Occludin and Claudin-1 protein expression was increased in the SJZT-L,SJZT-M,SJZT-H and MCA groups(P < 0.05),with a significant increase in the SJZT-H and MCA groups compared with the Model group.Conclusion:1.The application of Sijunzi decoction can reduce the content of Scr,BUN and IL-6 in the blood of CKD mice,protect renal function,delay renal pathological damage and alleviate intestinal pathological structure.2.Si Jun Zi Tang can reduce IS and TMAO content in serum of CKD mice,decrease UTs,increase intestinal tissue Claudin-1 and Occludin protein expression and reduce intestinal inflammatory factor IL-6 content,enhance tight junctions,repair intestinal barrier damage,reduce local inflammation in the intestine,and achieve the effect of improving intestinal barrier function in CKD mice.