摘要
目的 明确垂体肿瘤转化基因1(PTTG1)通过调控肠上皮细胞的焦亡水平在结肠炎症中的作用机制。方法 PTTG1野生型(WT)小鼠和PTTG1基因敲除(KO)小鼠各10只,随机分为4组,每组5只,分别为PTTG1 WT对照组(control组)和实验组(3%DSS组),PTTG1 KO对照组和实验组。实验组小鼠自由饮用3%葡聚糖硫酸钠(DSS)6 d诱导急性实验性结肠炎,对照组给予无菌双蒸水。观察并记录各组小鼠的疾病活动指数(DAI)。收集小鼠结肠组织,用免疫组织化学和蛋白免疫印迹法检测焦亡相关蛋白NLRP3、ASC、GSDMD的表达水平。在人源结肠上皮细胞系(HcoEpic)中,通过shRNA沉默PTTG1的表达,TNF-α刺激细胞以诱导细胞炎症模型,检测GSDMD的表达水平。结果 DSS诱导的结肠炎小鼠结肠黏膜组织中PTTG1表达减少(P <0.01),PTTG1敲除加重小鼠结肠炎症,PTTG1 KO实验组小鼠的结肠黏膜上皮焦亡相关蛋白表达水平上调(P <0.05)。在HcoEpic中沉默PTTG1的表达,TNF-α刺激后,细胞的GSDMD蛋白表达水平上调(P <0.05)。结论 PTTG1抑制肠上皮细胞的焦亡,当PTTG1缺失时,肠上皮细胞焦亡水平上调加重结肠炎。
Objective To investigate whether the pituitary tumor transformation gene 1(PTTG 1)plays a role in colitis by regulating intestinal epithelial cells pyroptosis.Methods Ten PTTG 1 wild-type(WT)mice and Ten PTTG 1 knockout(KO)mice were randomly divided into 4 groups of 5 each,respectively PTTG1 WT control and experimental group,PTTG1 KO control and experimental group.The mice in the experimental group were given 3%dextran sodium sulfate(DSS)for 6 days to induce acute colitis,and the control group was given sterile double distilled water(ddH2O).The disease activity index of the respective group of mice was observed and recorded.Mouse colonic tissue were collected,and the expression levels of NLRP3,ASC,and GSDMD were determined by immuno-histochemistry and western blot.In HCoEpiC,PTTG1 expression was knocked down using shRNA,and the cells were subsequently treated with TNF-αto induce inflammation.Then,the expression of GSDMD was detected.Results The expression of PTTG1 was decreased in colonic mucosal tissue in mice with acute colitis(P<0.01).Compared with WT mice,the colitis was significantly aggravated in PTTG1 KO mice after 3%DSS treatment.The expression of pyroptosis-related proteins was significantly up-regulated in the colon mucosal tissues of PTTG1 KO experimental mice(P<0.05).After knocking down the expression of PTTG1 in HCoEpiC and TNF-αtreatment,the expression levels of GSDMD were significantly up-regulated(P<0.05).Conclusion PTTG1 reduced pyroptosis in intestinal epithelial cells(IECs),while PTTG1 loss can enhance IEC pyroptosis,aggravating colonic inflammation.
作者
易玉君
翟晓明
刘慧玲
陶金
YI Yujun;ZHAI Xiaoming;LIU Huiling;TAO Jin(Department of Gastroenterology,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)
出处
《实用医学杂志》
CAS
北大核心
2024年第5期632-638,共7页
The Journal of Practical Medicine
基金
国家自然科学青年科学基金项目(编号:81800458)。
