| ObjectiveTo explore whether Huangqi-Honghua combination regulating intestinal flora to affect bile acid metabolism to protect cerebral injury,it could provide data support and theoretical basis for clinical application and drug development of Huangqi and Honghua.Methods1.Study on the pharmacodynamic effect of Huangqi-Honghua on cerebral ischemia-reperfusion injury: SD rats were randomly divided into control group(Sham),Model group(Model),Huangqi-Honghua group(HQ-HH)and Naoxintong capsule group(NXT),the rat middle cerebral artery occlusion/reperfusion(MCAO/R)model was established by suture method.After 72 hours of reperfusion,the pharmacodynamic effect of Huangqi-Honghua on cerebral ischemia-reperfusion injury was evaluated by neurological function,cerebral infarction volume and pathological section staining of brain tissue.2.The effects of Huangqi-Honghua on intestinal microflora of rats with cerebral ischemia reperfusion: 16 S r RNA sequencing was used to analyze the differences and changes of intestinal flora among groups.The effects of Huangqi-Honghua on the protein expressions of bile acid receptor FXR and intestinal barrier tight junction ZO-1,Occludin and claudin-1 in the small intestine of rats with cerebral ischemia reperfusion were investigated by Western blot experiment.3.The effects of Huangqi-Honghua on bile acid metabolism and brain protection in cerebral ischemia-reperfusion rats: based on metabonomics technology to detect bile acids in the feces of rats in each group,and the data were analyzed by principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA),comprehensive evaluation of Huangqi-Honghua bile acid metabolism of cerebral ischemia reperfusion rats.The expressions of IL17 A and Foxp3 in rat small intestine and Th17/Tregs cells in brain tissue were detected by immunofluorescence.Western blot was used to detect the expression of tight junction protein and IL17 A,IL10,IL6 and MMP9 in rat brain tissue.Results1.Huangqi-Honghua has a definite effect on improving cerebral ischemiareperfusion injury in rats.Compared with the model group,HQ-HH group and NXT group had good effect on improving neurological function and cerebral infarction volume in rats.The results of HE staining showed that Huangqi-Honghua could protect the morphology and structure of nerve cells and reduce the death of neurons.2.The 16 S r RNA sequencing analysis showed that there were significant differences in the structure of intestinal flora among the groups,which could cause changes in the structure of intestinal flora in rats after modeling.Huangqi-Honghua can regulate the relative abundance of intestinal flora back to the sham operation group,reduce the relative abundance of Ruminococcaceae,Bacteroides,Phascolarctobacterium and Desulfovibrionaceae,increase the relative abundance of Blautia,Lachnospiraceae,Oscillibacter and Bifidobacterium,regulate the homeostasis of intestinal flora.In this study,it was found that Huangqi-Honghua could also activate the expression of bile acid receptor FXR protein in the small intestine of rats and improve the intestinal barrier function.3.The content of bile acids in rat feces was detected by metabolomics,and the content of different bile acids was analyzed.The results showed that compared with the Sham group,the contents of Allo LCA(Allolithocholic acid),Iso LCA(Isolithocholic acid),Alpha-MCA(α-Muricholic acid),Beta-MCA(β-Muricholic acid),Beta-UDCA(3β-Ursodeoxycholic acid)and CA(Cholic acid)in the feces of rats in the model group were significantly increased(P < 0.05).After administration of Astragalus membranaceus and safflower,the contents of Allo LCA(Allolithocholic acid),Iso LCA(Isolithocholic acid),Beta-UDCA(3β-Ursodeoxycholic acid)and CA(Cholic acid)were decreased(P < 0.05).Huangqi-Honghua can regulate the differentiation of Th17 and Tregs cells in the small intestine of rats,reduce the expression of IL17 A in the small intestine of rats,reduce the transfer of IL17 A from the small intestine to the blood and then to the brain,and reduce brain damage.Astragalus-safflower can also regulate the differentiation of Th17 and Tregs cells in brain tissue,reduce the expression of pro-inflammatory factor IL17 A and increase the expression of anti-inflammatory factor IL10,reduce brain inflammation,protect the integrity of blood brain barrier and play a cerebral protection.Conclusion1.Huangqi-Honghua combination can significantly reduce the cerebral infarction in rats,improve the damaged nerve function and the morphology of nerve cells,and have a certain protective effect on cerebral ischemia reperfusion injury.2.Huangqi-Honghua combination can regulate intestinal intestinal flora,activate bile acid receptor FXR to regulate bile acid metabolism,reduce Th17 cells in rat brain tissue,increase Tregs cells,reduce the expression of pro-inflammatory factor IL17 A and increase the expression of anti-inflammatory factor IL10,and protect the integrity of blood-brain barrier,thereby improving cerebral ischemia-reperfusion injury. |
PDF Full Text Request