Study On The Mechanism Of Action Of Sijunzi Decoction In Improving Intestinal Microecology In CKD Mic

Purpose: In this study,an animal model of CKD was established by 5/6 nephrectomy,and the changes in intestinal flora in CKD mice were observed after treatment with Sijunzi decoction.Based on the theory of the "intestine-kidney axis",the influence of Sijunzi decoction on intestinal flora and its mechanism of protecting renal function were discussed.Material and method: Sixty C57BL/6 mice were randomly divided into sham operation group,model group,Meichang ’an group(225 mg/kg/d),and Sijunzi decoction low,medium,and high dose groups(2.34 g/kg/d,4.68 g/kg/d,9.36 g/kg/d).After the success of the model,the corresponding drug treatment was given for 8 weeks.After the experiment,the mice were killed and their serum was collected to detect the contents of creatinine Scr,BUN,PCS,IL-1β,and TNF-α.The contents of IL-1β and TNF-α mRNA in colon tissue were detected by RT-qCPR.Pathomorphological changes in the kidney and colon were observed by HE and IF staining.The protein expression of ZO-1 in colon tissue was detected by the WB method.Observe the intestinal contents of mice in each group,cultivate the flora,and observe the changes in the intestinal flora.Results:1.Serum levels of Scr,BUN,and PCS in mice: Compared with the sham-operated group,the serum levels of Scr,BUN,and PCS in the model group were significantly higher(P < 0.05,P < 0.01);Compared with the model group,the serum contents of Scr,BUN,and PCS decreased after treatment in Sijunzi decoction group and Meichang ’an group(P < 0.01).2.Effect of Sijunzi Decoction on renal morphology: Compared with the sham-operated group,the glomerulus in the model group was obviously atrophied,with severe mesangial cell proliferation and a large number of inflammatory cells infiltration;The lumen of the renal tubule is dilated,and the dilated lumen is folded.Epithelial cells are necrotic and exfoliated,and the renal tubule is vacuolated locally.After the intervention of the Sijunzi decoction group and Meichang ’an group,the pathological results of each group were improved.3.Effect of Sijunzi Decoction on inflammation(Elisa method): The levels of TNF-α and IL-1β in the model group were higher than those in the sham-operated group(P < 0.01).After the intervention of Sijunzi Decoction and Meichang ’an in different concentrations,the levels of TNF-α and IL-1β in each group decreased,except for the level of IL-1β in the low-dose group of Sijunzi Decoction,the other groups had the following characteristics.4.Effect of Sijunzi Decoction on the expression levels of TNF-α and IL-1β mRNA in colon tissue(RT-QCPR detection method): Compared with the sham operation,the levels of TNF-αand IL-1β mRNA in the colon of the model group increased significantly(P < 0.01);Compared with the model group,the levels of TNF-α and IL-1β mRNA in Sijunzi decoction group and Meichang ’an group decreased(P < 0.05,P < 0.01).5.Effect of Sijunzi Decoction on intestinal morphology: HE staining showed that the intestinal tissue structure of the sham operation group was complete,no obvious inflammatory cell infiltration was found,and there were neat intestinal villi;In the model group,intestinal villi were injured and a large number of inflammatory cells infiltrated;Compared with the model group,the pathological damage of colon tissue in Sijunzi decoction group and Meichang ’an group was significantly improved after intervention,especially in the high dose group.6.Effect of Sijunzi Decoction on the expression level of ZO-1 protein in colon tissue(WB detection method): Compared with sham operation group,the expression level of ZO-1 protein in model group decreased significantly(P < 0.01);However,after the intervention of Sijunzi decoction group and Meichang ’an group,the expression of ZO-1 protein in colon tissue was improved(P < 0.05,P < 0.01).7.Effect of Sijunzi Decoction on the expression level of ZO-1 protein in colon tissue(IF staining method): Compared with the sham operation group,the expression level of ZO-1 protein in the model group decreased significantly(P < 0.01);The expression of ZO-1 protein in colon tissue of mice was significantly increased after the intervention of Sijunzi decoction group and Meichang ’an group(P < 0.01).8.Effect of Sijunzi Decoction on intestinal flora: Compared with the sham operation group,the contents of Escherichia coli and Enterococcus in the model group increased,while the contents of Bifidobacterium and Lactobacillus decreased(P < 0.01);Compared with the model group,the contents of Escherichia coli and Enterococcus decreased(P < 0.05)and the contents of Bifidobacterium and Lactobacillus increased(P < 0.01)after the intervention of Sijunzi decoction group and Meichang ’an group.Conclusion:1.Sijunzi decoction can improve the kidney function and morphology of CKD mice and reduce the accumulation of urotoxin in CKD mice;2.Urotoxin can affect the inflammatory state of the body and increase the inflammatory factors in the intestine of mice through the intestinal barrier during CKD;3.The intestinal barrier of CKD mice is destroyed,which is closely related to tight junction protein ZO-1;Inflammatory cell infiltration in intestinal tissue is related to inflammatory factors TNF-α and IL-1β.4.The intestinal flora of mice with chronic kidney disease changed,which mainly showed that the contents of Escherichia coli and Enterococcus increased,while the contents of Bifidobacterium and Lactobacillus decreased;5.Sijunzi decoction can regulate intestinal flora through the "intestine-kidney axis",improve the intestinal barrier,reduce the micro-inflammatory state of the body and intestine,and play a role in protecting renal function.