背景:广泛的研究揭示了雷公藤红素(CEL)对各种疾病的有利作用,但CEL在自身免疫性葡萄膜炎中的作用仍有待研究.
方法:我们首先通过大鼠实验性自身免疫性葡萄膜炎模型评估CEL对自身免疫性葡萄膜炎的预防和治疗作用。在网络药理学之后,功能富集和分子对接分析,我们预测了CEL的潜在目标,并通过临床和组织病理学评分验证了其对EAU的影响,伊文思蓝染色,免疫荧光分析和蛋白质印迹。然后,我们通过16SrRNA测序和非靶向代谢组学分析评估了CEL在肠道环境中的作用。
结果:我们证实CEL治疗抑制了病理TH17反应,抑制炎症细胞的迁移,并通过靶向STAT3-IL17途径保持BRB的完整性。此外,通过粪便16SrRNA测序,发现CEL降低了机会致病菌的相对丰度,包括梭状芽胞杆菌1,副梭菌和GCA-900066575,并丰富了EAU大鼠中有益的盘旋菌和Ruminococcus扭矩群的相对丰度。同时,CEL治疗通过增加胆酸的相对浓度重塑EAU大鼠的肠道代谢产物,孕酮和guggulstone,降低异丙肾上腺素的相对水平,肌酐和苯乙酰谷氨酰胺.
结论:CEL通过靶向STAT3和重新描述肠道微环境的双重机制对实验性自身免疫性葡萄膜炎发挥改善作用。
BACKGROUND: Extensive research has revealed the favorable effects of celastrol (CEL) against various diseases, but the role of CEL in autoimmune uveitis remains unexplored.
METHODS: We first assessed the prophylactical and therapeutical effects of CEL on autoimmune uveitis via rat experimental autoimmune uveitis model. After network pharmacology, functional enrichment and molecular docking analyses, we predicted the potential target of CEL and validated its effect on
EAU by clinical and histopathological scores, Evans blue staining, immunofluorescence assay and western blotting. Then we evaluated the role of CEL in the gut environment by 16S rRNA sequencing and untargeted metabolomic analysis.
RESULTS: We confirmed that CEL treatment suppressed the pathological TH17 response, inhibited the migration of inflammatory cells, and preserved the integrity of BRB via targeting STAT3-IL17 pathway. Furthermore, CEL was found to reduce the relative abundance of opportunistic pathogenic bacteria including Clostridium_sensu_stricto_1, Parasutterella and GCA-900066575, and enrich the relative abundance of beneficial Oscillospirales and Ruminococcus_torques_group in
EAU rats by fecal 16S rRNA sequencing. Meanwhile, CEL treatment reshaped the gut metabolites in the
EAU rats by increasing the relative concentrations of cholic acid, progesterone and guggulsterone, and decreasing the relative levels of isoproterenol, creatinine and phenylacetylglutamine.
CONCLUSIONS: CEL exerts its ameliorative effects on the experimental autoimmune uveitis through the dual mechanisms of targeting STAT3 and reprofiling the gut microenvironment.