miR-2053在口腔黏膜修复中对成纤维细胞的影响及其机制研究

Effect of mi R-2053 on Fibroblasts in Oral Mucosal Repair and Its Mechanism

目的探索miR-2053在口腔黏膜修复中对成纤维细胞的影响及其作用机制。方法选择口腔黏膜病患者40例为实验组,健康者40例为对照组;采用实时荧光定量PCR检测口腔黏膜组织的miR-2053表达情况。建立人牙龈成纤维细胞转染miR-2053模型,采用CCK-8法检测细胞存活率,应用流式细胞仪检测细胞凋亡情况,Transwell小室法检测细胞迁移率;应用RT-PCR及Western-blot检测其组织重塑中的关键转录因子CollagenⅠ、CollagenⅢ、MMP-1、MMP-13的基因及蛋白表达水平。结果与对照组比较,实验组miR-2053在口腔黏膜组织中的表达明显升高(P<0.05);与对照组及阴性模拟物组比较,转染miR-2053的人牙龈成纤维细胞模型的细胞活力及迁移率均出现明显下降(P<0.05),而凋亡显著增高(P<0.05)。与对照组及阴性模拟物组比较,转染miR-2053后的人牙龈成纤维细胞中CollagenⅠ、CollagenⅢ、MMP-1、MMP-13基因的表达量均明显下降(P<0.05),CollagenⅠ、CollagenⅢ、MMP-13蛋白的相对表达量亦明显降低(P<0.05)。结论 miR-2053可能通过影响CollagenⅠ、CollagenⅢ、MMP-1、MMP-13的表达,进而影响人牙龈成纤维细胞的活力、凋亡及迁移能力,从而影响口腔黏膜病的康复及病程。

Objective To explore the effect of miR-2053 on fibroblasts in oral mucosal repair and its mechanism.Methods Forty patients with oral mucosal disease were selected as the case group, and 40 healthy subjects from the physical examination center were selected as the control group. Real-time quantitative PCR was used to verify the expression of miR-2053 in oral mucosa. The human gingival fibroblasts were transfected with miR-2053. The cell viability was detected by CCK-8 method. The cell mobilization rate was detected by flow cytometry and the cell migration rate was detected by Transwell chamber method. RT-PCR and Western-blot were used to detect the key transcription factors Collagen Ⅰ,Collagen Ⅲ, MMP-1, MMP-13 and their protein expression levels in tissue remodeling. Results Compared with the control group, the expression of miR-2053 in oral mucosa was significantly increased(P<0.05). Cell viability and migration rate of human gingival fibroblast model transfected with miR-2053 were both significantly lower than that of the control group and the negative analogue group(P <0.05), and the apoptosis was significantly increased(P <0.05). The expression levels of Collagen Ⅰ, Collagen Ⅲ, MMP-1 and MMP-13 in human gingival fibroblasts transfected with miR-2053 were significantly decreased compared with the control group and the negative analogue group(P <0.05). Further detection of protein levels revealed that the relative expression levels of Collagen Ⅰ, Collagen Ⅲ and MMP-13 in human gingival fibroblasts transfected with miR-2053 were significantly lower than those in the control group and the negative analogue group(P<0.05).Conclusion miR-2053 may affect the vitality, apoptosis and migration of human gingival fibroblasts by affecting the expression of Collagen Ⅰ, Collagen Ⅲ, MMP-1 and MMP-13, thus affect the rehabilitation of oral mucosal disease and the disease course.