CAJ | 학술논문

目的研究微小RNA(miRNA)-10a在IBD患者肠黏膜、血清及外周血单个核细胞(PBMC)中的表达,探讨其在疾病发生过程中的作用及意义.方法收集9例活动期UC患者、11例活动期CD患者及8名肠镜无异常者的内镜肠黏膜活组织检查标本,12例活动期UC患者、13例活动期CD患者及9名健康对照者的血清标本,9例活动期UC患者、11例活动期CD患者及8名健康对照者的PBMC标本,采用荧光定量PCR技术检测肠黏膜内、血清及PBMC中miRNA-10a表达以及肠黏膜内IL-12/IL-23 p40表达.收集活动期UC和CD患者各8例,取英夫利昔(IFX)治疗前和经IFX治疗3次后6周的肠黏膜检测miRNA-10a表达.另收集同期的11例活动期UC患者和10例活动期CD患者的肠黏膜活组织检查标本在体外使用IFX刺激培养18 h,检测肠黏膜细胞miRNA-10a表达.三样本比较采用单因素方差分析,两样本采用配对t检验,相关性分析采用Spearman检验.结果与健康对照组相比,UC及CD患者肠黏膜组织、血清、PBMC中miRNA-10a表达显著降低(F=38.45、30.46、14.74,P均＜0.05),UC及CD组间的表达量差异无统计学意义(P＞0.05),UC及CD患者肠黏膜组织IL-12/IL-23 p40表达明显升高(F=32.90,P＜0.05).CD患者结肠黏膜中IL-12/IL-23 p40与miRNA-10a表达呈负相关(r=-0.8545,P＜0.01).经IFX治疗3次后,IBD患者肠黏膜miRNA-10a表达明显升高(t=3.341、3,382,P均＜0.05).IFX体外刺激后肠黏膜miRNA 10a表达明显升高(t=3.095、7.193,P均＜0.05).结论 miRNA-10a与IBD患者炎性反应密切相关,其对IL-12/IL-23 p40有靶向作用,miRNA-10a可能成为IBD治疗新靶点.
목적 연구미소RNA(miRNA)-10a재IBD환자장점막、혈청급외주혈단개핵세포(PBMC)중적표체,탐토기재질병발생과정중적작용급의의.방법 수집9례활동기UC환자、11례활동기CD환자급8명장경무이상자적내경장점막활조직검사표본,12례활동기UC환자、13례활동기CD환자급9명건강대조자적혈청표본,9례활동기UC환자、11례활동기CD환자급8명건강대조자적PBMC표본,채용형광정량PCR기술검측장점막내、혈청급PBMC중miRNA-10a표체이급장점막내IL-12/IL-23 p40표체.수집활동기UC화CD환자각8례,취영부리석(IFX)치료전화경IFX치료3차후6주적장점막검측miRNA-10a표체.령수집동기적11례활동기UC환자화10례활동기CD환자적장점막활조직검사표본재체외사용IFX자격배양18 h,검측장점막세포miRNA-10a표체.삼양본비교채용단인소방차분석,량양본채용배대t검험,상관성분석채용Spearman검험.결과 여건강대조조상비,UC급CD환자장점막조직、혈청、PBMC중miRNA-10a표체현저강저(F=38.45、30.46、14.74,P균＜0.05),UC급CD조간적표체량차이무통계학의의(P＞0.05),UC급CD환자장점막조직IL-12/IL-23 p40표체명현승고(F=32.90,P＜0.05).CD환자결장점막중IL-12/IL-23 p40여miRNA-10a표체정부상관(r=-0.8545,P＜0.01).경IFX치료3차후,IBD환자장점막miRNA-10a표체명현승고(t=3.341、3,382,P균＜0.05).IFX체외자격후장점막miRNA 10a표체명현승고(t=3.095、7.193,P균＜0.05).결론 miRNA-10a여IBD환자염성반응밀절상관,기대IL-12/IL-23 p40유파향작용,miRNA-10a가능성위IBD치료신파점.
Objective To investigate the expression of microRNA (miRNA)-10a in the intestinal mucosa,serum and peripheral blood mononuclear cell (PBMC) of patients with inflammatory bowel disease (IBD) and explore its role and relevance in the pathogenesis of the disease.Methods The intestinal or colonic mucosal biopsy specimens of nine active ulcerative colitis (UC) patients,11 active Crohn's disease (CD) patients and eight patients with negative colonoscopy result as control were collected.The sera of 12 active UC patients,13 active CD patients and nine healthy controls were collected.The PBMC of nine active UC patients,11 active CD patients and eight healthy controls were collected.The expression of miRNA-10a in the intestinal mucosa,sera and PBMC and the expression of IL-12/IL-23 p40 in the intestinal mucosa were detected by real-time polymerase chain reaction (PCR).Each 8 cases of active UC and CD patients were collected.The intestinal mucosa before infliximab (IFX) treatment and six weeks after three times of IFX treatment were collected.And at same time,the intestinal mucosa of 11 active UC patients and 10 active CD patients were collected and cultured for 18 hours stimulated with IFX in vitro and then the expression of miRNA-10a in the intestinal mucosa was tested.One-way analysis of variance was used for comparison in three samples.Paired t-test was used for two samples comparison.Spearman test was used for correlation analysis.Results Compared with healthy controls,the expression of miRNA-10a in the intestinal mucosa,serum and PBMC of UC and CD patients significantly decreased (F=38.45,30.46 and 14.74,all P＜0.05).There was no statistic significance between UC and CD groups.The expression of IL-12/IL-23 p40 in the intestinal mucosa of UC and CD patients significantly increased (F=32.90,P＜0.05).The expression of IL-12/IL-23 p40 was negatively correlated with the expression of miRNA-10a in the intestinal mucosa of CD patients.After three times of IFX treatment,the expression of miR-10a in the intestinal mucosa of IBD patients significantly increased (t=3.341,3.382,both P＜0.05).After stimulated with IFX in vitro,the expression of miRNA-10a in the intestinal mucosa significantly increased (t=3.095,7.193,both P＜0.05).Conclusions miRNA-10a was closely correlated with the inflammation of IBD patients and with the role of targeting IL-12/IL-23 p40.miRNA-10a might be a new target for the IBD treatment.