民族药理学相关:白头翁汤(PD)是治疗溃疡性结肠炎的经典处方。先前的研究表明,PD的治疗效果与法尼醇X受体(FXR)的激活密切相关。FXR的活性受顶端钠依赖性胆汁酸转运蛋白(ASBT)的调节,和FXR-ASBT级联反应,以胆汁酸受体FXR为中心,胆汁酸代谢稳态对预防溃疡性结肠炎(UC)的发生和进展具有重要作用。研究目的:阐明PD对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎发挥蛋白活性作用的潜在机制,重点研究了FXR和ASBT的调制。材料与方法:建立急性溃疡性结肠炎模型,连续7天给予BALB/C小鼠在其饮用水中的3.5%DSS。采用疾病活动指数(DAI)评价各组小鼠的临床症状。使用糖原希夫高碘酸希夫(PAS)和阿尔辛蓝染色技术评估结肠组织中的杯状细胞表达。通过酶联免疫吸附试验(ELISA)检测血清和结肠组织中炎性细胞因子的表达。使用PCRArray芯片筛选了88个与PD治疗UC中FXR-ASBT途径相关的差异基因。进行蛋白质印迹(WB)分析以检测小鼠结肠组织中差异表达基因的蛋白质表达水平。结果:PD治疗有效降低了疾病活动指数(DAI)评分,减轻了结肠组织病理学损害,同时也恢复体重和结肠长度。此外,它显著减轻溃疡性结肠炎(UC)的严重程度,调节炎症,调节杯状细胞数量,并恢复胆汁酸平衡.此外,PCR阵列分析鉴定了21个与FXR-ASBT途径有关的差异表达基因。Westernblot结果显示FXR显著恢复,PD治疗后GPBAR1,CYP7A1和FGF15蛋白表达水平;此外,观察到ABCB11和RXRα表达水平增加的趋势。结论:PD治疗UC小鼠疗效显著,可能归因于其对胆汁酸稳态的调节,增强肠屏障功能,和肠道炎症的减弱。
Ethnopharmacological relevance: Pulsatilla decoction (PD) is a classical prescription for the treatment of ulcerative colitis. Previous studies have demonstrated that the therapeutic efficacy of PD is closely associated with the activation of Farnesoid X receptor (FXR). The activity of FXR is regulated by apical sodium-dependent bile acid transporter (ASBT), and the FXR-ASBT cascade reaction, centered around bile acid receptor FXR, plays a pivotal role in maintaining bile acid metabolic homeostasis to prevent the occurrence and progression of ulcerative colitis (UC). Aim of the study: To elucidate the underlying mechanism by which PD exerts its proteactive effects against Dextran Sulfate Sodium Salt (DSS)-induced ulcerative colitis, focusing on the modulation of FXR and ASBT. Materials and methods: To establish a model of acute ulcerative colitis, BALB/C mice were administered 3.5% DSS in their drinking water for consecutive 7 days. The disease activity index (DAI) was employed to evaluate the clinical symptoms exhibited by each group of mice. Goblet cell expression in colon tissue was assessed using glycogen schiff periodic acid-Schiff (PAS) and alcian blue staining techniques. Inflammatory cytokine expression in serum and colonic tissues was examined through enzyme-linked immunosorbent assay (ELISA). A PCR Array chip was utilized to screen 88 differential genes associated with the FXR-ASBT pathway in UC treatment with PD. Western blotting (WB) analysis was performed to detect protein expression levels of differentially expressed genes in mouse colon tissue. Results: The PD treatment effectively reduced the Disease Activity Index (DAI) score and mitigated colon histopathological damage, while also restoring weight and colon length. Furthermore, it significantly alleviated the severity of ulcerative colitis (UC), regulated inflammation, modulated goblet cell numbers, and restored bile acid balance. Additionally, a PCR Array analysis identified 21 differentially expressed genes involved in the FXR-ASBT pathway. Western blot results demonstrated significant restoration of FXR, GPBAR1, CYP7A1, and FGF15 protein expression levels following PD treatment; moreover, there was an observed tendency towards increased expression levels of ABCB11 and RXRα. Conclusion: The therapeutic efficacy of PD in UC mice is notable, potentially attributed to its modulation of bile acid homeostasis, enhancement of gut barrier function, and attenuation of intestinal inflammation.