miR-199抑制增生性瘢痕的机制研究

Study on mechanism of miR-199 inhibiting hypertrophic scar

目的探讨mi R-199抑制增生性瘢痕中的作用机制。方法自2018年10月至2019年3月,选取北部战区总医院烧伤整形外科就诊的8例增生性瘢痕患者的组织作为观察组,8例皮瓣移植手术中弃用的正常皮肤组织作为对照组。通过实时荧光定量PRC分析正常皮肤组织和增生性瘢痕皮肤组织中miR-199的表达情况。培养原代增生性瘢痕成纤维细胞,分别使用无意义小链、mi R-199和miR-199抑制剂转染。噻唑蓝实验检测不同处理因素作用0、12、24、36、48 h时mi R-199对细胞增殖的影响。Hoechst 33258染色检测mi R-199对细胞凋亡的影响。Transwell实验检测miR-199对细胞迁移的影响。结果实时荧光定量PRC显示,与对照组比较观察组中存在mi R-199表达下调(P<0.05)。噻唑蓝实验显示,miR-199能够抑制成纤维细胞的增殖(P<0.05)。Hoechst 33258染色实验显示,miR-199可以促进成纤维细胞的凋亡。Transwell实验显示,miR-199可以抑制成纤维细胞的迁移。结论 mi R-199在增生性瘢痕中表达下调,能够抑制增生性瘢痕成纤维细胞的增殖,促进了凋亡并且抑制了迁移,从而抑制瘢痕的增生。

Objective To investigate the mechanism of miR-199 inhibiting hypertrophic scar. Methods From October 2018 to March 2019, the hypertrophic scars from 8 patients were selected as observation group and normal skin from 8 patients were selected as control group. The expression of mi R-199 in normal skin and hypertrophic scar was analyzed by real-time PCR. The primary hypertrophic scar fibroblasts were cultured and transfected with nonsense chains, miR-199 and mi R-199 inhibitor. The effect of mi R-199 on proliferation of hypertrophic scar fibroblastsat 0, 12, 24, 36 and 48 h was detected by thiazolyl blue assay. The effect of mi R-199 on apoptosis of hypertrophic scar fibroblasts was analyzed by Hoechst 33258 staining. The effect of mi R-199 on migration of hypertrophic scar fibroblasts was detected by transwell assay. Results Compared with control group, real-time PCR showed that the expression of miR-199 was down regulated in observation group(P<0.05). Thiazolyl blue assay showed that the proliferation of fibroblastscould be inhibited by mi R-199(P<0.05). Hoechst 33258 staining showed that the apoptosis of fibroblasts could be promoted by miR-199. Transwell assay showed that the migration of fibroblasts could be inhibited by mi R-199. Conclusion The down-regulation of mi R-199 expression in hypertrophic scar could inhibit the proliferation and migrationof hypertrophic scar fibroblasts and promote the apoptosis.