四神丸通过干预miRNA200/ZEB信号通路调控记忆性Th1/Th2治疗溃疡性结肠炎的作用机制研究
Mechanism of Sishen Pills on treating ulcerative colitis by modulating memory Th1/Th2 via miRNA200/ZEB signaling pathway
黄佳琦 1龙健 2赵海梅 3刘端勇4
作者信息
- 1. 江西中医药大学研究生院,南昌 330004
- 2. 江西中医药大学临床医学院,南昌 330004
- 3. 江西中医药大学中医学院,南昌 330004
- 4. 江西中医药大学护理学院,南昌 330004
- 折叠
摘要
目的:从miRNA200/ZEB信号,探讨四神丸有效治疗溃疡性结肠炎的可能机制.方法:将BALB/c小鼠随机分成正常组(Normal组)、正常+四神丸组(Nor+SSP组)、模型组(DSS组)、模型+四神丸组(DSS+SSP组),采用葡萄糖硫酸钠(DSS)建立溃疡性结肠炎小鼠模型,四神丸给药14d后,通过体质量、疾病活动评分、病理损伤评分等评价其疗效;流式细胞术检测小鼠记忆性Th1(mTh1)和Th2(mTh2)细胞水平,实时荧光定量(RT-PCR)法检测结肠中miRNA200家族和ZEB信号通路关键基因的表达,用免疫蛋白印迹法(WB)检测ZEB1和ZEB2.结果:与DSS组比较,经四神丸干预后,小鼠体质量、脾脏和结肠各项数据恢复,疾病活动评分和病理损伤评分降低,mTh1和Th2细胞水平明显下降(P＜0.01,P＜0.05),结肠中miRNA200家族和ZEB1的mRNA水平上升(P＜0.05,P＜0.01),ZEB2mRNA水平下降(P＜0.01),ZEB1、ZEB2蛋白水平显著下调(P＜0.01).结论:四神丸能有效缓解小鼠溃疡性结肠炎,可能是通过抑制miRNA200/ZEB信号调控记忆性Th1/Th2细胞亚群实现的.
Abstract
Objective:Investigate the mechanism of SSP treating ulcerative colitis(UC)via miRNA200/ZEB signaling pathway.Methods:BALB/c mice were randomly divided into Normal,Nor+SSP,DSS,and DSS+SSP groups.UC model was established using DSS.After 14 d of administration of SSP,the changes of body weight,DAI score,and pathological injury score were used to evaluate the curative effect.The levels of memory Th1 and Th2 cell were detected by flow cytometry,the levels of miRNA200 family and key genes of ZEB signaling pathway in colon were detected by RT-PCR.ZEB1 and ZEB2 protein levels were detected by Western Blot(WB).Results:Compared with DSS group,after the intervention of SSP,the colon length was significantly increased,indexes of spleen and colon were recovered,the pathological injury scores were decreased,the levels of memory Th1 and Th2 cells were significantly decreased(P＜0.01,P＜0.05),and levels of miRNA200 family and ZEB1 mRNA were significantly recovered(P＜0.01,P＜0.05).ZEB2 mRNA levels were significantly decreased(P＜0.01),and SSP significantly down-regulated ZEB1 and ZEB2 protein levels(P＜0.01).Conclusion:SSP relieved UC in mice,which may be achieved by inhibiting miRNA200/ZEB signal to regulate memory Th1/Th2 cells.
关键词
四神丸/溃疡性结肠炎/miRNA200/ZEB/记忆性Th1/Th2/作用机制Key words
Sishen Pills/Ulcerative colitis/miRNA200/ZEB/Memory Th1/Th2/Mechanism引用本文复制引用
基金项目
国家自然科学基金(82060799)
江西省研究生创新专项(研字[2020]39号)
出版年
2024
NETL
NSTL
万方数据