白藜芦醇对瘢痕疙瘩成纤维细胞自噬的影响

Effect of resveratrol on autophagy in keloid fibroblasts

目的观察白藜芦醇（Res）对瘢痕疙瘩成纤维细胞自噬的影响，探讨Res抗瘢痕疙瘩增生机制。方法取手术切除瘢痕疙瘩组织成纤维细胞原代培养，将瘢痕疙瘩成纤维细胞分为空白对照组和用含不同浓度（20、40、80、160μmol／L）Res实验组，给药24、48h后噻唑蓝（MTT）法检测细胞活性，实时荧光定量聚合酶链反应（FQ-PCR）检测微管相关蛋白1轻链3（LC3）-Ⅱ及Beclin1mRNA表达，酶联免疫吸附试验（ELISA）法检测不同浓度Res对瘢痕疙瘩成纤维细胞自噬相关蛋白LC3-Ⅱ及Beclin1合成的影响。结果Res显著抑制瘢痕疙瘩成纤维细胞增殖，且存在一定的时间和浓度依赖性，随着浓度升高和时间延长，其抑制率由4．9％增至43．7％；Res作用后瘢痕疙瘩成纤维细胞自噬相关蛋白LC3-Ⅱ及Beclin1mRNA相对表达量增加，分别从（1．367±0．141）升至（4．562±0．371）倍，（1．232±0．189）升至（3．603±0．297）倍。自噬相关蛋白LC3-Ⅱ及Beclinl的吸光度（A）值分别从0．211±0．017升至0．872±0．052，0．171±0．021升至0．544±0．028。结论Res有效促进自噬相关蛋白LC3-Ⅱ及Beclin1mRNA及蛋白表达，提高瘢痕疙瘩成纤维细胞自噬水平，抑制瘢痕疙瘩成纤维细胞增殖。

Objective To observe autophagy of keloid fibroblasts induced by resveratrol and explore the possible mechanism. Methods The cultured in vitro fibroblasts from patients with keloid were harvested and divided into control group and resveratrol （20, 40, 80, 160 μ mol/L） groups. After treatment for 24, and 48 h, cell activity was determined by methyl thiazol tetrazolium （MTT） assay. Real -time fluorescent quantitative polymerase chain reaction （ FQ - PCR） was used to examine the expression level of microtu- bule - associated protein 1 light chain 3 （ LC3 ） - Ⅱ and Becline 1. Enzyme linked immunosorbent assay （ELISA） was used to detect the expression level of LC3 - Ⅱand Becline 1. Results Resveratrol could significantly inhibit the growth of fibroblasts in vitro in a concentration - and time - dependent manner, and the inhibition rate increased from 4.9% to 43.7% with increase of concentration and prolongation of time. After treatment, keloid fibroblasts autophagy - related protein LC3 - Ⅱ and beclinl mRNA relative ex- pression increased, from （ 1. 367 ± 0. 141 ） to （4. 562 ± 0. 371 ） times, and （ 1. 232 ± 0. 189） to （3. 603 ± 0. 297） times, respectively. Conclusion Resveratrol can effectively promote the production of protein LC3 - Ⅱ and Beclin 1, increase the level of autophagy in keloid fibroblasts, and inhibit the growth of fibroblasts.