| Objective: To explore schisandrin B(Sch B)reduces intestinal ischemia Reperfusion(IIRI)injury and the nuclear factor of NF-E2 related factor 2(Nrf2)/heme oxygenase 1(HO-1)signaling pathway and apoptosis in mice by network pharmacology and in vivo experiment.Methods: The targets of Sch B were searched from Pharm Mapper,CTD,HERB,Pubchem,Swiss Target Prediction,ZINC and the targets related to intestinal IIRI injury from Dis Ge NET,Gene Cards.Using R language to draw Venn diagram to obtain the common target of Sch B and intestinal IIRI injury,and the protein-protein interaction(PPI)network and "intestinal IIRI injury-Sch B-key target gene" network were established based on STRING and Cytoscape,followed by Gene Oncology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.36 healthy male SPF C57BL/6J mice were divided into 6 groups by random number table method(n=6): Sham operation group(group S),sham operation + schisandrin B group(group S+Sch B),intestinal ischemia reperfusion group(group IIRI),IIRI+Sch B group,IIRI+Nrf2 inhibitor ML385group(IIRI+ML385 group),and IIRI+Sch B+ML385 group(SMB group).By clamping superior mesenteric artery,the model of intestinal IIRI injury was established for 45 min,and reperfusion for 2 h in anesthetized mice.Sch B 80 mg/kg was administered with intragastrically for 7 consecutive days in Sch B groups,and ML385 30 mg/kg was intraperitoneally injected with 1h prior to ischemia in ML385 mice.Mice were sacrificed at 2 h of reperfusion,intestine tissues were took and observed the morphological structure under light microscope after stained by HE,and the injury was performed via the Chiu’s score,and the apoptosis index was observed and calcμLated via TUNEL staining,and the levels of Nrf2 and HO-1 protein were detected via immunohistochemistry staining,and Nrf2,HO-1and apoptosis related proteins(such as Bax,Bcl-2 and cleaved caspase 3)were determined by Western blot.Results: A total of 412 Sch B related targets,2166 intestinal IIRI injury related targets(The figure is represented by IIRI)and 153 common targets were screened out.PPI analysis showed 88 core target genes included NFE2L2(Nrf2),HMOX1(HO-1),BCL2,CASP3(caspase 3),etc.KEGG enrichment analysis screened 163 related pathways,showing that apoptosis pathway may play a key role in the treatment of intestinal IIRI injury by Sch B.The animal experiment have shown that,the Chiu’s score and apoptosis index were increased,expression of Nrf2,HO-1,Bax,cleaved caspase 3 were up-regμLated,expression of Bcl-2 and Bcl-2/Bax were down-regμLated in IIRI group compared to S group and S+Sch B group(P<0.05).The Chiu’s score and apoptosis index were decreased,expression of Nrf2,HO-1,Bcl-2 and Bcl-2/Bax were up-regμLated,expression of Bax,cleaved caspase 3were down-regμLated in IIRI+Sch B group compared to IIRI group(P<0.05),and the above indexes in IIRI+Sch B group were opposite to those in IIRI+ML385 group(P<0.05),and the above indexes in SMB group basically recovered to the level of IIRI group.The Chiu’s score,apoptosis index and expression of Nrf2,HO-1,Bcl-2 and Bcl-2/Bax were down-regμLated,expression of Bax,cleaved caspase 3 were up-regμLated in SMB group compared to IIRI group(P<0.05).Conclusion: This study reveals that Sch B has the characteristics of mμLti-target and mμLti-pathway in the reduction of intestinal IIRI injury,and Sch B can reduces intestinal IIRI injury through inhibiting apoptosis by activation of Nrf2/HO-1 pathway. |
PDF Full Text Request