Role of lincRNA-p21 in alcohol-induced intestinal injury
Objective To investigate the role of intergenic long non-coding RNA-p21(lincRNA-p21)in alcohol-induced intestinal injury.Methods C57BL/6Cnc mice were fed adaptively for one week,and we randomly selected 10 mice and randomly divided them into the lincRNA-p21 overexpression group and control group,with 5 mice in each group.The mice in the above two groups were injected with lincRNA-p21-over-AAV8-U6-GFP virus and Control-AAV8-U6-GFP virus through the tail vein,respectively.Five Trp53cor1 knockout C57BL/6 mice(lincRNA-p21-/-)were used as KO group,and five C57BL/6Cnc mice were used as wild control group(WT group).Each group was fed a liquid diet contain-ing 5%alcohol for 4 weeks,and 40%alcohol was given 5 g/kg for the last 3 days to establish the alcohol injury models.After the successful modeling,the mice were killed and the intestinal tissues were retained.The expression of lincRNA-p21 in the intestinal tissues was detected by RT-qPCR.The pathological morphological changes of intestinal tissue were observed by HE staining.The expression levels of claudin-1 and ZO-1 in the intestinal tissues were detected by immunohistochemis-try.Microbial DNA was extracted from cecum contents of mice,and 16S amplicon sequencing was performed to analyze the polymorphism and composition of microflora.Results The relative expression level of lincRNA-p21 in intestinal tis-sues of the overexpression group was higher than that of the control group(t=5.31,P<0.05).The relative expression of lincRNA-p21 in the KO group was lower than that in the WT group(t=3.95,P<0.01).HE staining showed that intestinal injury and infiltration of granulocyte and monocyte in intestinal mucosa were observed in all groups.Compared with the control group,the intestinal injury of lincRNA-p21 overexpression group was more serious,intestinal villi vacuolation increased,intestinal mucosal cell structure was more disordered,intestinal wall edema and intestinal wall recess damage were more serious.Compared with the WT group,intestinal villi vacuolation was reduced in the KO group,intestinal mucosal cell structure was more orderly,intestinal wall edema and intestinal wall recess injury were less severe.The rela-tive expression levels of claudin-1 and ZO-1 in the lincRNA-p21 overexpression group were lower than those in the control group(t =20.84 and 7.57,respectively;both P<0.01).The relative expression levels of claudin-1 and ZO-1 in the KO group were higher than those in the WT group(t=5.53 and 6.69,respectively;both P<0.01).The intestinal flora sequencing results showed that the dominant intestinal flora of the lincRNA-p21 overexpression group and control group,KO group and WT group were firmicutes or bacteroides.The abundance of actinomycetes in intestinal flora of the lincRNA-p21 overexpression group was higher than that of the control group(t=4.21,P<0.01).The abundance of cyanobacteria in the intestinal flora of the KO group was lower than that of the WT group(t=2.62,P<0.05).KEGG analysis showed that the effects of lincRNA-p21 overexpression and gene knockout on intestinal microflora function in alcohol-exposed mice were related to amino acid metabolism,lipid metabolism,nutrient digestion and absorption,and cell growth and death,etc.Conclusions Overexpression of lincRNA-p21 can exacerbate alcoholic intestinal injury,while knockdown of lincRNA-p21 can alleviate alcoholic intestinal injury.The mechanism may be related to increasing intestinal barrier damage and altering intestinal microbiota structure.
alcoholic intestinal injuryintergenic long non-coding RNA-p21intestinal barrierintestinal flora
万方数据
维普
