| Purpose of research:Acute kidney injury(AKI)is a common and critical emergency,and ischemia and reperfusion injury(IRI)is one of the common causes of AKI,more than half of the patients will gradually progress to chronic kidney disease(CKD).With the incidence and mortality of the disease increasing year by year,it is urgent to find the effective treatment drug for the disease.The study found that patients who with kidney disease had enhanced oxidative stress and intestinal dysbiosis.Shenshuai Xiezhuo decoction has been put into clinical use for many years.Previous studies have demonstrated that Shenshuai Xiezhuo decoction can significantly improve renal function in patients with CKD,inhibit oxidative stress and improve the structure of intestinal flora.Starting from oxidative stress and intestinal flora,our research group further explored the pathway regulation mechanism of renal function,renal tissue and intestinal histopathology,oxidative stress,intestinal flora and kidney injury related proteins in AKI rats,so as to provide experimental basis for the scientific prevention and control of AKI in Shenshuai Xiezhuo decoction.Methods:A total of 24 SD rats were selected and randomly divided into sham,model,low-,medium-,and high-dose groups of Shenshuai Xiezhuo decoction and emodin group.In all groups except the sham group,the bilateral renal pedicles were occluded by micro artery clamping for 45 min to establish acute renal ischemia-reperfusion model in rats.The model and sham operated groups were intragastrically administrated with normal saline,and the other groups were intragastrically administrated with corresponding doses for 15 d,respectively.The total 24-h urinary protein,serum creatinine(Cre)and blood urea nitrogen(BUN)levels in urine of rats in each group were detected by ELISA;PCR was used to detect the distribution of Escherichia coli,Bacteroides fragilis and Enterococcus faecalis in the feces of medium dose group in Shenshuai Xiezhuo decoction;HE staining was used to detect the histopathological damage of renal tissue and small intestine;The expression of apoptosis inhibitor of macrophage(AIM),kidney injury molecule 1(KIM-1)in kidney tissue was detected by immunohistochemistry;KIM-1,serum D-lactate and neutropil gelatinase-associated lipocalin(NGAL)content in urine,and serum D-lactate levels were measured by ELISA;The expression levels of AIM,KIM-1,phospho-c-Jun N-terminal kinases(p-JNK),and phospho-p38mitogen-activated protein kinases(p-p38)proteins in renal tissues were determined by Western blot.Results:1.Renal function results of rats in each group: The levels of BUN,Cre and urinary protein in the model group and emodin group were significantly increased compared with those in the sham group(P < 0.05);Compared with the model group,the urine protein content of Cre and urinary protein at 1d and 14 d in the low dose group of Shenshuai Xiezhuo decoction significantly decreased(P < 0.05),BUN,Cre and urinary protein content were significantly decreased in the middle-dose grope and high dose group(P <0.05),Urinary protein content was significantly lower in the emodin group at 1d and 7d(P <0.05),while there was no significant difference in BUN,Cre and urinary protein content at 14 d in the emodin group.2.Results of renal histopathological injury of rats in each group:In the model group,the renal tubules were loosely arranged,and the tissue was edematous,with damage to the glomerular structure;In the high-dose group and emodin group of Shenshuai Xiezhuo decoction,the renal tubules were tightly arranged,the glomerular structure was better,and edema was reduced.3.Results of renal tubular injury-related protein in each group: By ELISA,compared with the sham grope,the grope with drug urinary NGAL was significantly increased at 7d and 14d(P <0.05),and KIM-1 protein content was significantly increased at 1d,7d and 14d(P <0.05);compared with the model grope,NGAL and KIM-1 content decreased in the group with drug,the medium dose group was the most significant(P <0.05).The WB test found significantly higher expression of AIM and KIM-1 protein in the model group compared with the sham group(P <0.05),and the expression of AIM and KIM-1 protein was significantly decreased in the high dose group and emodin group compared with the model group(P <0.05).Immunohistochemical testing found that AIM and KIM-1 expression were increased in the model group compared with the sham group;AIM and KIM-1 expression were decreased in the group with drug compared with the model group,and AIM and KIM-1 protein expression were the least in the high dose group and emodin group.4.Results of intestinal histopathological injury in each group:Structural injury of intestinal glands,edema of small intestine muscle layer and ring muscle tissue in model group;In the low,medium and high dose groups,the intestinal glands were arranged neatly,and the edema of the annular muscle tissue was reduced;The emodin group arranged intestinal glands.5.Structure of intestinal flora and intestinal mucosal permeability of rats in the medium dose group of renal failure decoction: PCR found that the medium dose group decreased at 14 d compared with the 1d(P <0.05);the beneficial B.fragilis and E.faecalis colonies increased significantly on 14 d compared with the 1d(P <0.05);ELISA found that serum D-lactate was significantly higher in the model group compared with the sham group(P <0.05);compared with the model group,serum D-lactate was significantly lower in the low and emodin group(P <0.05).6.Results of p-p38 and p-JNK protein expression measured by Western Blot: Both p-p38 and p-JNK protein were significantly upregulated in the model group compared with the sham group(P <0.05);p-p38 and p-JNK protein were significantly decreased in the low-,medium-,high-dose group compared with the model group(P <0.05),among them,the medium dose group and emodin group had a better effect on p-p38 protein,while the high dose group had a better effect on p-JNK protein.Conclusion:1.Shenshuai Xiezhuo decoction improved renal function in AKI rats.2.Shenshuai Xiezhuo decoction may improve renal pathological damage in AKI rats.3.Shenshuai Xiezhuo decoction can alleviate AKI by downregulating NGAL,AIM and KIM-1 proteins.4.Shenshuai Xiezhuo decoction improved the intestinal pathological damage in AKI rats.5.Shenshuai Xiezhuo decoction improved the intestinal flora structure and reduced intestinal mucosal permeability in AKI rats.6.Shenshuai Xiezhuo decoction can regulate the expression of p-p38 and p-JNK proteins and inhibit oxidative stress. |
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