摘要
目的:通过对溃疡性结肠炎(ulcerative colitis,UC)患者外周血单个核细胞进行内质网应激(endoplasmic reticulum stress,ERS)刺激,比较X-盒结合蛋白1(X-box binding protein 1,XBP1)剪切形式(XBP1 splicing,XBP1s)及炎性细胞因子表达水平变化,探讨UC患者对ERS反应的变化.方法:通过实时聚合酶链反应(real timepolymerase chain reaction,RT-RCR)检测XBP1s,实时定量聚合酶链反应(quantitative-polymerase chain reaction,Q-PCR)检测白介素-2(interleukin-2,IL-2)、γ-干扰素(interferon-γ,IFN-γ)、IL-17α表达变化.比较UC之间差异并进行相关分析.结果:(1)用植物凝集素(phytohemagglutinin,PHA)及毒胡萝卜素(thapsigargin,TG)单独及联合刺激正常人及UC患者外周血单个核细胞后XBP1均发生剪切,联合刺激时剪切形势较明显;(2)分别PHA与TG、联合刺激正常人及UC患者外周血单个核细胞,发现联合刺激组IL-2、IFN-γ、IL-17α表达水平增高.结论:在ERS状态下,正常人群及UC患者的T淋巴细胞均可通过XBP1s所介导的信号通路对ERS发生响应,激活炎性细胞因子表达,加重炎性发生,与正常人相比,UC患者外周血单个核细胞对ERS作用更为敏感.
AIM： To demonstrate the regulatory effect of endoplasmic reticulum stress（ERS） on T lymphocyte function and the pathogenesis of ulcerative colitis（UC） by stimulating peripheralblood mononuclear cells of UC patients with thapsigargin（TG） and measuring the splicing of X-box binding protein 1（XBP1） and changes of cytokine expression. METHODS： Primary T lymphocytes were seeded in 12-well plates and stimulated with TG for 12 h. The cells were collected and lysed for m RNA analysis of cytokine expression and XBP1 splicing. All subjects were from Department of Gastroenterology of Chinese PLA 309 th Hospital from May 2002 to March 2013, and UC was diagnosed according to the diagnostic standard for UC which was formulated in 2007 in Jinan. Peripheral blood mononuclear cells were isolated, seeded in 12-well plates and stimulated with phytohemagglutinin（PHA）, TG, or PHA ＋ TG, respectively. The expression of interleukin-2（IL-2）, interferon-γ（IFN-γ）, and IL-7α was analyzed by quantitative polymerase chain reaction（Q-PCR）, and the splicing of XBP1 was detected by real-time polymerase chain reaction（RT-PCR）.RESULTS： XBP splicing was detected when peripheral blood mononuclear cells were stimulated with PHA and TG, alone or in combination, both in UC patients and healthy adults, and XBP splicing was more significant in cells from UC patients treated with PHA plus TG. Increased levels of IL-2, IFN-γ, and IL-17α could be observed both in UC patients and healthy adults when peripheral blood mononuclear cells were stimulated with PHA plus TG, and the increase was more significant in the UC group.CONCLUSION： Compared to health subjects,UC patients are more sensitive to ERS in terms of increased XBP 1 splicing and up-regulation of inflammatory cytokines such as IL-2, IFN-γ and IL-17α.
出处
《世界华人消化杂志》
CAS
2015年第8期1359-1368,共10页
World Chinese Journal of Digestology
基金
北京市自然科学基金资助项目
No.7132175~~
关键词
内质网应激
溃疡性结肠炎
X-盒结合蛋白1
Endoplasmic reticulum stress
Ulcerative colitis
X-box binding protein 1
