肿瘤抑制因子在溃疡性结肠炎肠黏膜的表达及其对NF-κB信号通路的影响  

The expression of CYLD in the intestinal mucosa of ulcerative colitis and its impact on the NF-κB signaling pathway

作　　者：雷媛[1] 陈刚[1] 欧士钰[1] 柳娟[1] 杜凌[1] Lei Yuan;Chen Gang;Ou Shijue(Department of Gastroenterology,the Fourth Affiliated Hospital of Guangxi Medical University,Liuzhou 545000,China)

机构地区：[1]广西医科大学第四附属医院消化内科,柳州545000

出　　处：《中华保健医学杂志》2023年第6期685-687,共3页Chinese Journal of Health Care and Medicine

基　　金：广西壮族自治区卫生健康委员会自筹经费科研课题(Z20200230)。

摘　　要：目的观察肿瘤抑制因子Cylindromatosis(CYLD)在溃疡性结肠炎患者肠黏膜组织中的表达水平,探讨其对NF-κB通路P65、P50表达的影响。方法选取2021年5月~2023年2月广西医科大学第四附属医院收治的60例溃疡性结肠炎(UC)患者肠黏膜标本(UC组),并选取同期30名正常者为健康对照组。采用RT-PCR法检测两组受检者组织中CYLD mRNA表达。以CYLD shRNA转染结肠Caco-2细胞株,Western blot法检测NF-κB信号通路P65、P50蛋白的表达以及加入NF-κB信号通路抑制剂吡咯烷二硫氨基甲酸(PTDC)后P65、P50蛋白的表达。结果与健康对照组比较,UC组mRNA表达水平明显降低[(0.624±0.087)vs.(0.993±0.056)],组间差异有统计学意义(t=21.119,P<0.05)。UC亚组间比较,重度组mRNA表达(0.418±0.059)低于中度和轻度组[(0.690±0.103)、(0.722±0.076)],差异有统计学意义(P<0.05),轻度、中度组间比较差异无统计学意义(P>0.05)。转染CYLD shRNA的结肠Caco-2细胞CYLD蛋白表达下调,P65、P50蛋白表达上调;加入PTDC后,P65、P50表达下降,进一步证实CYLD能够负调控NF-κB信号通路。结论溃疡性结肠炎患者肠黏膜CYLD表达下调,CYLD可能通过负调控NF-κB信号通路影响UC的发病过程。Objective To observe the expression level of tumor suppressor factor Cylindromatosis(CYLD)in intestinal mucosa tissue of patients with ulcerative colitis,and explore its impact on the expression of NF-KB pathway P65 and P50.Methods Collect intestinal mucosal samples from 60 patients with ulcerative colitis(UC),and select 30 normal individuals as the control group during the same period admitted to our hospital from May 2021 to Feburary 2023.RT-PCR method was used to detect the expression of CYLD mRNA in tissues.Colon Caco-2 cell line was transfected with CYLD shRNA,and Western blot was used to detect the expression of P65 and P50 proteins in the NF-KB signaling pathway,as well as the expression of P65 and P50 proteins after the addition of the NF-KB signaling pathway inhibitor pyrrolidine dithiocarbamate(PTDC).Results Compared with normal colon tissue,the mRNA expression level in the UC group was significantly reduced[(0.624±0.087)vs.(0.993±0.056)],and the difference between the groups was statistically significant(t=21.119,P<0.05).Compared between UC subgroups,the mRNA expression in the severe group(0.418±0.059)was lower than that in the moderate and mild groups[(0.690±0.103),(0.722±0.076)](P<0.05).There was no statistically significant difference between the mild and moderate groups(P>0.05).The expression of CYLD protein was downregulated in colon Caco-2 cells transfected with CYLD shRNA,while the expression of P65 and P50 proteins was upregulated.After the addition of PTDC,the expression of P65 and P50 decreased,further confirming that CYLD can negatively regulate the NF-KB signaling pathway.Conclusion The expression of CYLD in the intestinal mucosa of patients with ulcerative colitis is downregulated,and CYLD may affect the pathogenesis of UC by negatively regulating the NF-KB signaling pathway.

关 键 词：肿瘤抑制因子 溃疡性结肠炎 核因子-ΚB 黏膜屏障 

分 类 号：R574.62[医药卫生—消化系统]