机构地区:[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 [2]空军军医大学基础医学院六大队,陕西西安710032
出 处:《中国美容整形外科杂志》2024年第7期395-398,共4页Chinese Journal of Aesthetic and Plastic Surgery
基 金:陕西省重点研发计划项目(2022SF-399)。
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(hypertrophic 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2μmol/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(CollagenⅠ)和α-平滑肌肌动蛋白(α-SMA)的m RNA表达水平差异,采用蛋白质印迹法检测HSF中CollagenⅠ和α-SMA的蛋白表达水平差异;采用免疫荧光法检测两组CollagenⅠ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CollagenⅠ与α-SMA的m RNA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CollagenⅠ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,CollagenⅠ与α-SMA主要定位在HSF的细胞质,SRT1720处理组CollagenⅠ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CollagenⅠ与α-SMA表达,从而抑制细胞外基质(extracellular matrix,ECM)合成与成纤维细胞的异常转分化。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 2μmol/L for 24 h.The mRNA expressions of collagen type 1(CollagenⅠ)andα-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 CollagenⅠandα-SMA in HSF were detected by Western blotting.The intracellular expression and localization of Colla-genⅠandα-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 CollagenⅠandα-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 CollagenⅠandα-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 CollagenⅠandα-SMA were mainly localized in the cytoplasm of HSF,and the expression of CollagenⅠandα-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 CollagenⅠandα-SMA in scar fibroblasts by activating SIRT1,thereby inhibiting extracellular matrix(ECM
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