SRT1720对瘢痕成纤维细胞的调控作用
The phenotypic regulation mechanism of SRT1720 on hypertrophic scar fibroblasts
严艺文 1张婷 2郑朝2
作者信息
- 1. 空军军医大学第一附属医院烧伤与皮肤外科,陕西西安 710032;空军军医大学基础医学院六大队,陕西西安 710032
- 2. 空军军医大学第一附属医院烧伤与皮肤外科,陕西西安 710032
- 折叠
摘要
目的 研究沉默信息调节因子1(silence information regulator 1,SIRT1)的激动剂SRT1720对增生性瘢痕成纤维细胞表型的调控作用.方法 自2022年1-6月,空军军医大学第一附属医院烧伤与皮肤外科将瘢痕真皮成纤维细胞(hyper-trophic scar fibroblasts,HSF)按随机数字表法分为PBS对照组与SRT1720处理组,样本数9个/组.先分别用等量PBS与终浓度为2 μmol/L的SRT1720处理HSF 24 h,然后分别采用实时荧光定量反转录PCR(RT-PCR)法检测HSF中Ⅰ型胶原蛋白(Collagen Ⅰ)和α-平滑肌肌动蛋白(α-SMA)的mRNA表达水平差异,采用蛋白质印迹法检测HSF中Collagen Ⅰ和α-SMA的蛋白表达水平差异;采用免疫荧光法检测两组Collagen Ⅰ和α-SMA在HSF中的胞内表达与定位情况.以上3种细胞实验中每组样本数均为3个.采用GraphPad Prim 8.0进行数据统计分析,组间数据比较采用独立样本t检验,P＜0.05为差异有统计学意义.结果 SRT1720处理组HSF中Collagen Ⅰ与α-SMA的mRNA表达量分别为(0.34±0.06)、(0.43±0.08),均明显低于 PBS 对照组(1.00±0.12)、(1.00±0.13),t 值分别为 8.57、6.79,P＜0.01;经 SRT1720 处理 HSF 的 Collagen Ⅰ 与 α-SMA 的蛋白表达量(0.70±0.05)、(0.64±0.09),均明显少于 PBS 对照组的(1.14±0.04)、(1.10±0.05),t 值分别为 12.61、7.37,P＜0.01.免疫荧光染色显示,Collagen Ⅰ与α-SMA主要定位在HSF的细胞质,SRT1720处理组Collagen Ⅰ与α-SMA的表达量较PBS对照组减少.结论 SRT1720能够通过激活SIRT1显著抑制瘢痕成纤维细胞Collagen Ⅰ与α-SMA表达,从而抑制细胞外基质(extra-cellular matrix,ECM)合成与成纤维细胞的异常转分化.
Abstract
Objective To explore the phenotypic regulation mechanism of silence information regulator 1(SIRT1)agonist SRT1720 on hypertrophic scar fibroblasts.Methods From January to June 2022.Department of Burns and Cutaneous Surgery,The First Affiliated Hospital of the Air Force Medical University,according to random number table method,hypertrophic scar fibroblasts(HSF)were divided into PBS control group and SRT1720-treated group.The two groups were respectively treated with equal amounts of PBS and SRT1720 with a final concentration of 2 μmol/L for 24 h.The mRNA expressions of collagen type 1(Collagen Ⅰ)and α-smooth muscle actin(α-SMA)in HSF were detected by a real-time fluorescence quantitative reverse transcription PCR(RT-PCR).The protein ex-pressions of Collagen Ⅰ and α-SMA in HSF were detected by Western blotting.The intracellular expression and localization of Colla-gen Ⅰ and α-SMA of each group in HSF were detected using immunofluorescence method.Statistical analysis was performed using GraphPad Prim 8.0,the data was analyzed by independent sample t test,and statistically significant differences were considered at P＜0.05.Results The mRNA expressions of Collagen Ⅰ and α-SMA in HSF in the SRT1720-treated group was(0.34±0.06)and(0.43±0.08),respectively,which were significantly lower than that in the negative control group of(1.00±0.12)and(1.00±0.13),t=8.57 and 6.79,P＜0.01.The protein expression of Collagen Ⅰ and α-SMA in HSF in the SRT1720-treated group was(0.70±0.05)and(0.64±0.09),which were significantly lower than that in the negative control group of(1.14±0.04)and(1.10±0.05),t=12.61 and 7.37,P＜0.01).Immunofluores-cence staining showed that Collagen Ⅰ and α-SMA were mainly localized in the cytoplasm of HSF,and the expression of Collagen Ⅰ andα-SMA in the SRT1720-treated group was reduced compared with that in the negative control group.Conclusion SRT1720 could sig-nificantly inhibit the expression of Collagen Ⅰ and α-SMA in scar fibroblasts by activating SIRT1,thereby inhibiting extracellular matrix(ECM)synthesis and abnormal transdifferentiation of fibroblasts.
关键词
瘢痕成纤维细胞/Ⅰ型胶原蛋白/α-平滑肌肌动蛋白Key words
Hypertrophic scar fibroblasts/Collagen Ⅰ/α-Smooth muscle actin引用本文复制引用
基金项目
陕西省重点研发计划项目(2022SF-399)
出版年
2024
NETL
NSTL
万方数据