MiR-506-3p与自噬相关因子在烧伤后皮肤成纤维细胞中的表达研究

Expression of MiR-506-3p and Autophagy-related Factors in Post-burn Skin Fibroblasts

目的:分析烧伤组织中miR-506-3p的表达及其对自噬水平的调控。方法:提取正常皮肤组织、烧伤皮肤组织和热处理成纤维细胞细胞的总RNA,采用荧光定量RT-qPCR、Western Blot分别检测各细胞miR-506-3p和自噬相关因子(LC3-Ⅱ/Ⅰ比值、Atg5、p62及CollagenⅠ)表达水平;采用同样方法检测miR-506-3p模拟物或miR-506-3p抑制剂转染到真皮成纤维细胞中对上述自噬相关因子表达水平的影响,并利用CCK-8方法检测转染细胞的增殖效率。结果:与正常组织相比,烧伤组织和热处理成纤维细胞中,miR-506-3p的表达下调,自噬相关因子LC3-Ⅱ/Ⅰ比值、Atg5及CollagenⅠ的表达显著上调(P<0.01),而p62表达下调(P<0.05);miR-506-3p抑制剂转染到皮肤成纤维细胞中,miR-506-3p表达显著降低,促进了细胞增殖和自噬发生,而miR-506-3p的过表达则显示出相反的作用(P<0.05)。结论:miR-506-3p在烧伤皮肤组织恢复中发挥着积极作用,对调节成纤维细胞的自噬水平具有重要作用,有可能成为烧伤愈合后瘢痕潜在治疗的靶标。

Objective To analyze the expression of miR-506-3p in post-burn tissues and its regulation of autophagy.Methods To extract the total RNA of normal skin tissue,burned tissue and heat treatment into fibroblast cells,using fluorescent quantitative RT-QPCR,Western Blot,respectively,detects mIR-506-3p and autophagy-related factors(LC3-Ⅱ/Ⅰ ratio,Atg5,p62 and Collagen Ⅰ) in post-burn tissue and heat-damaged skin fibroblasts,RT-qPCR was also used to detect the effect of miR-506-3p mimic or miR-506-3p inhibitor transfected into dermal fibroblasts on the expression of autophagy-related factors,and CCK-8 method was used to detect the proliferation efficiency of transfected cells.Results Compared with normal tissues,in post-burn tissues and heat-treated fibroblasts,the expression of miR-506-3p was down-regulated,and the expression of autophagy-related factors,such as LC3-Ⅱ/Ⅰ ratio,Atg5 and Collagen I,were significantly up-regulated,while the expression of p62 was down-regulated.When miR-506-3p inhibitors were transfected into dermal fibroblasts,the expression of miR-506-3p was significantly down-regulated,which could promote cell proliferation and autophagy,while the overexpression of miR-506-3p showed the opposite effect.Conclusion MiR-506-3p plays an positive role in the recovery of post-burn skin,and plays an important role in regulating the autophagy level of fibroblasts,it may be a potential target for post-burn scar treatment.