miR-21对瘢痕疙瘩成纤维细胞胶原分泌及细胞增殖的影响

目的 探讨mi R-21(micro RNA-21)在瘢痕疙瘩中的表达及其生物学作用,为瘢痕疙瘩的防治提供新的思路。方法 收集临床患者正常皮肤及瘢痕疙瘩标本,部分进行组织学检测;部分进行体外细胞培养,Real-time PCR检测体外培养的正常皮肤来源和瘢痕疙瘩来源的成纤维细胞中,mi R-21、Smad7(mi R-21靶基因)、Col 1 A1、Col 3 A1(纤维化相关基因)的表达;应用mi RNA-21的模拟物和抑制剂,以Western-Blot和Real-time PCR分别检测Col 1 A1、Col 3 A1及Smad7的蛋白和核酸水平表达变化,结晶紫实验检测细胞增殖能力的变化。结果 瘢痕疙瘩标本组织学检测,其胶原的含量明显高于正常皮肤;瘢痕疙瘩成纤维细胞中mi R-21、Col 1 A1、Col 3 A1表达高于正常皮肤组织,Smad7表达低于正常皮肤组织;正常皮肤来源成纤维细胞转染mi R-21模拟物后,Smad7表达降低,纤维化相关基因的表达、细胞增殖能力增加;瘢痕疙瘩组织来源成纤维细胞转染mi R-21抑制剂后,Smad7表达增加,纤维化相关基因的表达、细胞增殖能力降低。结论 瘢痕疙瘩中mi R-21表达升高,促进了细胞外基质中纤维化相关胶原基因的表达,促进成纤维细胞的体外增殖能力,可能是导致瘢痕疙瘩形成的重要原因。

The Effect of miR-21 on Collagen Synthesis and Cell Proliferation of Fibroblasts in Keloid

Objective To investigate the expression and function of miR-21 (microRNA-21) in the formation of the keloid, and provide a new direction for the prevention and treatment of the scar. Methods Normal skin tissue and keloid were harvested for histological detection and cell culture in vitro. Real-time PCR was used to detect the expression of mir21, Smad7, Col 1 A1 and Col 3 A1 in normal and keloid fibroblasts. The expression of Smad7, Col 1 A1 and Col 3 A1 with miR-21 mimics and anti-miR-21 oligonucleotides were also detected by Western-Blot and Real-time PCR. The cell proliferation was also detected by crystal violet assay. Results The content of collagen was significantly higher in keloid tissue than that of normal skin. The expression of miR-21, Col 1 A1 and Col 3 A1 were higher in keloid fibroblasts than that of normal fibroblasts, while the expression of Smad7 was lower. After transfection, the expression of Smad7 was decreased, the expression of scar related genes and cell proliferation were increased in normal fibroblasts. And the contrast results were observed in keloid fibroblasts after transfection. Conclusion The high expression of miR-21 can increase the expression of scar related genes and promote the proliferation of fibroblasts in vitro, which may be an important cause of the formation of the scar.