BACKGROUND Inflammatory bowel disease(IBD)affects millions of people worldwide and has emerged as a growing problem in industrialized nations.The lack of therapeutic targets has limited the treatment of IBD.Studies fo...BACKGROUND Inflammatory bowel disease(IBD)affects millions of people worldwide and has emerged as a growing problem in industrialized nations.The lack of therapeutic targets has limited the treatment of IBD.Studies found that parasitic nematode infections can ameliorate clinical and experimental colitis.Our previous study found that rSj16,a 16-kDa secreted protein of Schistosoma japonicum produced by Escherichia coli,has protective effects on dextran sulfate sodium(DSS)-induced colitis in mice.Apoptosis is an important factor in the pathogenesis of colitis.However,it is not clear whether the effect of rSj16 on colitis is related to apoptosis.AIM To investigate whether the protective effects of rSj16 on colitis is related to apoptosis and its mechanism.METHODS In-vivo,colitis was induced by DSS.The severity of colitis was assessed.WB was used to detect the changes of apoptosis-related genes in colon tissues.Q-PCR was used to detect the changes of miRNA-217-5p and HNF1B.In-vitro,WB was used to detect the changes of apoptosis-related genes in intestinal epithelial cells.TUNNEL staining and flow cytometry were used to detect cell apoptosis.RESULTS rSj16 attenuates clinical activity in DSS-induced colitis mice.TUNNEL staining and WB results showed that apoptosis was increased in colon tissue after treatment with DSS,and the apoptosis of colon tissue was significantly reduced after treatment with rSj16.Compared with normal mice,the expression of miR-217-5p was increased in colon tissue of DSS-induced colitis mice.In addition,the miR-217-5p target gene hnf1b was decreased after administration of DSS.After treatment with rSj16,the expression of miR-217-5p was decreased and the expression of HNF1B was increased compared with the DSS-treated group.When Etoposide was used in combination with miR-217-5p mimic on MODE-K cells,the expression of cleaved-Caspase-3 and Bax was increased,and Bcl-2 was decreased compared with only Etoposide treatment,the expression of HNF1B was significantly reduced,suggesting that miR-217-5p acts as a pro-apoptotic in colon epithelial cells and down-regulates the target gene hnf1b.After rSj16 administration in MODE-K cells,miR-217-5p expression was significantly decreased,HNF1B expression was increased,and apoptosis was reduced.CONCLUSION The protective effects of rSj16 on colitis is related to apoptosis and miRNA-217-5p may be a further target for therapeutic intervention against IBD.展开更多
基金The study was reviewed and approved by the Sun Yat-sen University Institutional Review Board.(Approval No.SYSU-IACUC-2019-B517)the National Natural Science Foundation of China,No.81902081 and No.81871682+5 种基金the Natural Science Foundation of Guangdong Province,No.2020A1515011573 and No.2019A1515012068China Postdoctoral Science Foundation,No.2018M640858 and No.2019T120771Fundamental Research Funds for the Central Universities,No.19ykpy170,No.17ykpy09 and No.19ykpy29National Science and Technology Major Project,No.2018ZX10101002-001the 111 Project,No.B12003and the Natural Science Foundation of Guangdong Province,No.2021A1515010976.
文摘BACKGROUND Inflammatory bowel disease(IBD)affects millions of people worldwide and has emerged as a growing problem in industrialized nations.The lack of therapeutic targets has limited the treatment of IBD.Studies found that parasitic nematode infections can ameliorate clinical and experimental colitis.Our previous study found that rSj16,a 16-kDa secreted protein of Schistosoma japonicum produced by Escherichia coli,has protective effects on dextran sulfate sodium(DSS)-induced colitis in mice.Apoptosis is an important factor in the pathogenesis of colitis.However,it is not clear whether the effect of rSj16 on colitis is related to apoptosis.AIM To investigate whether the protective effects of rSj16 on colitis is related to apoptosis and its mechanism.METHODS In-vivo,colitis was induced by DSS.The severity of colitis was assessed.WB was used to detect the changes of apoptosis-related genes in colon tissues.Q-PCR was used to detect the changes of miRNA-217-5p and HNF1B.In-vitro,WB was used to detect the changes of apoptosis-related genes in intestinal epithelial cells.TUNNEL staining and flow cytometry were used to detect cell apoptosis.RESULTS rSj16 attenuates clinical activity in DSS-induced colitis mice.TUNNEL staining and WB results showed that apoptosis was increased in colon tissue after treatment with DSS,and the apoptosis of colon tissue was significantly reduced after treatment with rSj16.Compared with normal mice,the expression of miR-217-5p was increased in colon tissue of DSS-induced colitis mice.In addition,the miR-217-5p target gene hnf1b was decreased after administration of DSS.After treatment with rSj16,the expression of miR-217-5p was decreased and the expression of HNF1B was increased compared with the DSS-treated group.When Etoposide was used in combination with miR-217-5p mimic on MODE-K cells,the expression of cleaved-Caspase-3 and Bax was increased,and Bcl-2 was decreased compared with only Etoposide treatment,the expression of HNF1B was significantly reduced,suggesting that miR-217-5p acts as a pro-apoptotic in colon epithelial cells and down-regulates the target gene hnf1b.After rSj16 administration in MODE-K cells,miR-217-5p expression was significantly decreased,HNF1B expression was increased,and apoptosis was reduced.CONCLUSION The protective effects of rSj16 on colitis is related to apoptosis and miRNA-217-5p may be a further target for therapeutic intervention against IBD.
