摘要
目的:探讨微小RNA-98-5p(miR-98-5p)和DNA甲基转移酶3A(DNMT3A)在溃疡性结肠炎(UC)患者和动物模型中的表达及作用机制.方法:选取2021年1月-2022年12月100例UC患者作为观察组,另选取100名健康受试者作为对照组,检测血清miR-98-5p、DNMT3AmRNA及蛋白表达水平,分析miR-98-5p、DNMT3A mRNA与UC临床病理特征的关系;建立UC大鼠模型,大鼠随机分为空白组(CT组)、模型组(UC组)、miR-98-5p对照组(antagomiR-NC组)、miR-98-5p抑制剂组(antagomiR-98-5p组),ELISA测定血清白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-a含量,HE染色观察结肠组织病理学变化,RT-qPCR和Western blot检测结肠组织miR-98-5p、DNMT3A mRNA及蛋白表达水平.结果:与对照组比较,UC患者血清miR-98-5p表达升高,DNMT3A mRNA、DNMT3A表达降低,miR-98-5p、DNMT3A mRNA表达水平与病情分级、黏液脓血便、黏膜急慢性炎症和不典型增生有关(P＜0.05);与CT组比较,UC组大鼠结肠黏膜层缺损,可见大量炎性细胞浸润,血清IL-1β、IL-6、TNF-α以及结肠组织miR-98-5p表达升高,DNMT3AmRNA及蛋白表达降低(P＜0.05);与antagomiR-NC组比较,antagomiR-98-5p组黏膜层缺损部分和炎症细胞浸润减少,结构较清晰,血清IL-1β、IL-6、TNF-α以及结肠组织miR-98-5p表达降低,DNMT3A mRNA及蛋白表达升高(P＜0.05);miR-98-5p与DNMT3A有靶向结合位点.结论:UC患者血清和模型大鼠结肠组织miR-98-5p表达升高,DNMT3A表达降低,下调miR-98-5p的表达能够促进DNMT3A的表达,改善UC症状.
Abstract
Objective:To investigate the expression and mechanism of microRNA-98-5p(miR-98-5p)and DNA methyltransferase 3A(DNMT3A)in patients with ulcerative colitis(UC)and animal model of ulcerative colitis.Methods:One-hundred UC patients in our hospital from January 2021 to December 2022 were collected as the observation group,another 100 healthy subjects were collected as the control group,serum miR-98-5p,DNMT3A mRNA and protein ex-pression were detected,the relationship between miR-98-5p,DNMT3A mRNA and clinical pathological characteristics of UC was analyzed;the UC rat model was established and randomly grouped into a blank group(CT group),a model group(UC group),a miR-98-5p control group(antagomiR-NC group),and a miR-98-5p inhibitor group(antagomiR-98-5p group),ELISA was applied to measure the levels of serum interleukin-1(IL-1),IL-6,and tumor necrosis factor(TNF),HE staining was applied to observe the histopathology changes of the colon,RT-qPCR and Western blot were applied to detect miR-98-5p,DNMT3A mRNA and protein expression levels in colon tissue.Results:Compared with the control group,the serum miR-98-5p expression of UC patients was increased,DNMT3A mRNA and DNMT3A were decreased,the expression of miR-98-5p and DNMT3A mRNA were associated with disease grading,mucopurulent bloody stools,mucosal acute and chronic inflammation,and atypical hyperplasia(P＜0.05);compared with the CT group,the colon mucosal layer of rats in the UC group showed defects,and obvious infil-tration of inflammatory cells,serum IL-1,IL-6,TNF and colon miR-98-5p expression were in-creased,the DNMT3A mRNA and protein expression were decreased(P＜0.05);compared with the antagomiR-NC group,the mucosal layer defect and inflammatory cell infiltration in the an-tagomiR-98-5p group were reduced,and the structure was clearer,serum IL-1,IL-6,TNF,and colon miR-98-5p expression were decreased,DNMT3A mRNA and protein expression were in-creased(P＜0.05);miR-98-5p had a targeted binding site with DNMT3A.Conclusion:Serum miR-98-5p expression of UC patients and the colon tissue of model rats was increased,while DNMT3A expression was decreased.Down-regulating miR-98-5p expression can promote the expression of DNMT3A and improve UC symptoms.
维普
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