盐酸小檗碱抑制NLRP3介导的细胞焦亡改善DSS诱导的大鼠溃疡性结肠炎  

Berberine hydrochloride ameliorates DSS-induced ulcerative colitis in rat by inhibiting pyroptosis mediated by NLRP3

作　　者：关玉龙[1] 张乐[1] 孔德润[2] GUAN Yulong;ZHANG Le;KONG Derun(Department of Gastroenterology,the Third People's Hospital of Bengbu,Bengbu 233000,China;Department of Gastroenterology,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽省蚌埠市第三人民医院消化内科,蚌埠233000 [2]安徽医科大学第一附属医院消化内科,合肥230022

出　　处：《中国免疫学杂志》2024年第9期1865-1870,共6页Chinese Journal of Immunology

摘　　要：目的:盐酸小檗碱(BBR)是治疗溃疡性结肠炎(UC)的常用药,但其挽救炎症反应导致的细胞焦亡的分子机制有待进一步研究。方法:SD雄性大鼠随机分为4组:对照组(Ctrl)、DSS模型组、BBR治疗组(DSS+BBR)、BBR+NLRP3激动剂组(DSS+BBR+BMS-986299)。采用右旋葡聚糖硫酸钠法(DSS)构建UC大鼠模型,同时BBR治疗组和BBR+NLRP3激动剂组每日灌胃BBR 2次,连续7 d,BBR+NLRP3组每日2次腹腔注射BMS-986299。实验期间每天称量体质量、观察小鼠一般情况并评估疾病活动指数(DAI);实验结束后解剖观察并评估结肠大体形态、测量长度;组织切片HE染色,光镜下观察结肠组织的病理学变化并评估组织损伤指数(TDI);ELISA检测结肠组织中细胞因子TNF-α和IL-1β含量,Western blot检测NLRP3、ASC、GSD?MD-N、Cleaved-caspase 1和IL-1β的表达。结果:BBR能够有效改善UC引起的体质量减轻、结肠的组织完整性、细胞焦亡,并降低结肠中TNF-ɑ、IL-1β等炎症因子的表达、焦亡细胞的比例以及NLRP3、IL-1β、GSDMD-N、Cleaved-caspase 1等细胞焦亡通路中关键蛋白的表达,并且BBR的治疗效果和调节表达的蛋白因NLRP3激活而逆转。结论:BBR通过降低细胞焦亡通路中NLRP3、IL-1β、GSDMD-N、Cleaved-caspase 1等蛋白的成熟体表达,发挥治疗效果,为BBR治疗UC提供了分子依据。Objective:Berberine hydrochloride(BBR)is commonly used for the treatment of ulcerative colitis(UC),but its molecular mechanism,especially the mechanism of rescue of inflammatory-induced pyroptosis,needs to be further analyzed.Meth⁃ods:SD male rats were randomly divided into four groups:control group(Ctrl),DSS model group,BBR treatment group(DSS+BBR)and BBR+NLRP3 agonist group(DSS+BBR+BMS-986299).The UC rat model was established by the dextran sulfate sodium method(DSS).At the same time,BBR was intragastrically administered to BBR treatment group and BBR+NLRP3 agonist group twice perday for 7 consecutive days.BMS-986299 was intraperitoneally injected to BBR+NLRP3 group twice a day.During the experiment,the body weights were weighed,the general condition was observed,and the disease activity index(DAI)was evaluated every day.After the experiment,the gross morphology and length of colon were observed and evaluated.HE staining was used to observe the pathological changes of colon tissue and evaluate the tissue injury index(TDI).ELISA was employed for detecting the contents of TNF-αand IL-1βin colon tissue,and Western blot was employed for detecting NLRP3,ASC,GSDMD-N,Cleaved-caspase 1 and IL-1βexpressions.Results:BBR could effectively improve the UC-induced weight loss,colon tissue integrity and cell pyroptosis,reduce the expression levels of inflammatory factors such as TNF-ɑand IL-1β,the proportion of pyroptosis,and the levels of proteins NLRP3,IL-1β,GSDMD-N,and Cleaved-caspase 1 which play crucial roles in pyroptosis pathway.The therapeutic effect and the regulated protein expression through BBR were reversed by NLRP3 activation.Conclusion:BBR exerts a therapeutic effect by reducing the proteins levels of mature NLRP3,IL-1β,GSDMD-N,Cleaved-caspase 1,which are responsible for pyroptosis pathway.This result provides a molecular basis for BBR in treatment of UC.

关 键 词：盐酸小檗碱 细胞焦亡 溃疡性结肠炎 NLRP3 

分 类 号：R285.5[医药卫生—中药学]