| Objective:To investigate the protective effect of Akkermansia muciniphila(A.muciniphila)on renal function in rats with adenine-induced chronic kidney disease(CKD).The mechanism may be related to the improvement of intestinal barrier by A.muciniphila.Methods:Culture A.muciniphila strain;Twenty six week old SD rats,half male and half female,were randomly divided into three groups:normal group(8 rats),CKD group(8rats)and experimental group(10 rats).Normal group:intragastric administration of 0.9%sodium chloride injection 1ml/d for 4 weeks;CKD group:gavage adenine 200mg/(kg·d)for 4 weeks;Experimental group:Adenine 200mg/(kg·d)and A.muciniphila1.5×10~9CFU/d were given by gavage at the same time.During the experiment,the mental state of the rats was observed,and the body weight was measured every week.After 4 weeks,the rats were anesthetized by intraperitoneal injection of 10%chloral hydrate,and the rat serum was collected by cardiac puncture.The rats were incised in the lower abdomen.The left kidney tissue was excised and weighed,and the colon tissue of the same segment was frozen at-80℃for examination.Serum of rats was collected to measure serum creatinine(Scr)and blood urea nitrogen(BUN),and renal function was assessed.After completion,the blood of rats was collected,the serum creatinine(Scr)and blood urea nitrogen(BUN)were measured,the glomerular filtration rate was calculated,and the renal function was evaluated.Masson staining was used to observe the interstitial fibrosis of renal tissue,and Periodic Acid-Schiff stain(PAS)staining was used to observe the thickness of renal tubular basement membrane in renal tissue.Western blot was used to detect the expression of tight junction proteins ZO-1,Occludin and Claudin-1 in colon tissue.At the same time,immunohistochemical staining was used to observe the expression of tight junction protein in rat colon tissue.PAS staining was used to observe the thickness of mucus layer and the number of goblet cells in colon tissue.Results:Compared with the normal group,the serum creatinine and blood urea nitrogen of the adenine-induced CKD group were increased(P<0.01),and the serum creatinine and blood urea nitrogen of the experimental group were lower than those of the CKD group(P<0.01).The results of Masson staining and PAS staining of kidney tissue showed that,compared with the normal group,the renal tissue of the CKD group had severe interstitial fibrosis,dilated renal tubules,and partial inflammatory cell infiltration(P<0.01).The renal interstitial fibrosis and renal tubular dilatation of the rats in the experimental group intervened by A.muciniphila were reduced(P<0.05).The results of Western blot and immunohistochemistry in colon tissue showed that the expressions of tight junction proteins ZO-1,Occludin and Claudin-1 in the CKD group were lower than those in the normal group(P<0.05),while those in the experimental group treated with A.The expression levels of the above proteins in mice were higher than those in the CKD group(P<0.05).The results of PAS staining of colon tissue showed that compared with the normal group,the colon glands of the CKD group were atrophied and deformed,the thickness of the mucus layer was significantly reduced,and the goblet cell count was significantly reduced(P<0.01).The thickness of mucus layer in colon tissue increased(P<0.05),and the number of goblet cells increased(P<0.01).Conclusion:A.muciniphila can improve renal function and renal interstitial fibrosis in rats with chronic kidney disease,and the mechanism may be related to the protection of intestinal barrier function by A.muciniphila. |
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