目的 探索泛凋亡在溃疡性结肠炎(UC)发病机制中的作用，研究香连丸对UC小鼠泛凋亡途径相关蛋白的调节机制。方法 48只C57BL/6小鼠随机分为6组:正常组、模型组、香连丸低、中、高剂量组及美沙拉嗪组。除正常组外，其余5组分别予3％葡聚糖硫酸钠(DSS)自由饮用1周，以制备实验性UC小鼠模型。从造模第1天开始，相应各组分别予美沙拉嗪或者香连丸灌胃，连续治疗1周。第8天，取各组小鼠结肠组织，取部分固定后用于苏木精-伊红(HE)染色进行组织病理观察，采用TUNEL法检测细胞凋亡情况，酶联免疫吸附(ELISA)实验检测IFN-γ、TNF-α、IL-1β和IL-6的含量，采用 Western blot 检测 Bax、Bcl-2、Caspase-8、p-MLKL、p-RIPK3、Caspase-1、GSDMD-N 和 IL-1β 蛋白的表达。结果 与模型组比较，香连丸高剂量组UC小鼠疾病活动指数(DAI)、结肠组织损伤程度、IFN-γ、TNF-α、IL-1β和IL-6的水平显著降低(均P＜0。01)，结肠长度及体重增加(均P＜0。01)。香连丸通过下调UC小鼠Bax/Bcl-2比值抑制凋亡(P＜0。01)，通过升高Caspase-8蛋白水平、降低p-MLKL、p-RIPK3蛋白水平抑制坏死性凋亡(均P＜0。01)，及通过降低Caspase-1、GSDMD-N和IL-1β蛋白表达水平抑制焦亡(均P＜0。01)。结论 香连丸可以从凋亡、坏死性凋亡、焦亡三方面综合调控泛凋亡途径，缓解DSS诱导的小鼠溃疡性结肠炎。
Effect of Xianglian Pills on Panapoptotic Pathway in Ulcerative Colitis Mice
Objective To explore the role of PANoptosis in the pathogenesis of ulcerative colitis(UC),and to study the regulatory mechanism of Xianglian pills on panapoptotic pathway related proteins in UC mice.Methods Forty-eight C57BL/6 mice were randomly divided into 6 groups:normal group,model group,mesalazine group,low,medium and high dose of Xiangli-an pills group.Except the normal group,the other 5 groups were given 3％dextran sulfate(DSS)for 1 week,respectively,to pre-pare experimental UC mice model.At the same time,starting from the first day of modeling,the corresponding groups were giv-en mesalazine or Xianglian pills by intragastric administration,respectively,for continuous treatment for 1 week.On the 8th day,colon tissues of mice in each group were taken and partially fixed for HE staining and pathological observation.TUNEL method was used to detect the apoptosis of colon tissue cells,and the contents of IFN-γ,TNF-α,IL-1β and IL-6 in colon were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the expression of Bax,Bcl-2,Caspase-8,p-MLKL,p-RIPK3,Caspase-1,GSDMD-N and IL-1β in colon tissues.Results Compared with model group,disease activity index(DAI)and degree of colon tissue damage decreased in high-dose Xianglian pills group(both P＜0.01),and the levels of IFN-γ,TNF-α,IL-1β and IL-6 in UC mice decreased(all P＜0.01),and colon lengths and body weights increase(both P＜0.01).In ad-dition,apoptosis of UC mice was inhibited in the high-dose group by downregulating Bax/Bcl-2 ratio(P＜0.01),and necroptosis was inhibited by increasing Caspase-8 protein level and decreasing p-MLKL and p-RIPK3 protein levels(all P＜0.01).Pyropto-sis was inhibited by decreasing the expression levels of Caspase-1,GSDMD-N and IL-1β(all P＜0.01).Conclusion Xianglian pills can relieve DSS induced ulcerative colitis in mice by regulating apoptosis,necroptosis and pyroptosis.
万方数据
维普
