العنوان البديل: Shikonin inhibits fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p. (English)

المؤلفون: 唐玉婷, 贺 茜, 万 瑀, 王建军, 杨安宁, 吴 凯, 焦 运, 白志刚, 姜怡邓, 沈江涌

المصدر: Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 12/18/2023, Vol. 27 Issue 35, p5642-5648, 7p

مصطلحات موضوعية: HYPERTROPHIC scars, SCARS, DIMETHYL sulfone, GENE expression, CELL migration, CELL culture, COLLAGEN

الملخص (بالإنجليزية): BACKGROUND: Previous studies have shown that shikonin has the potential to treat hypertrophic scar and that microRNAs are involved in the regulation of the pathological mechanism of hypertrophic scar. It is speculated that shikonin may regulate the occurrence and development of hypertrophic scar through microRNAs regulation. OBJECTIVE: To investigate the mechanism of shikonin on the fibrosis of human hypertrophic scar fibroblasts via MicroRNA-382-5p. METHODS: Hypertrophic scar tissue and normal skin tissue adjacent to the scar (within 3 cm around the scar) were provided by the Department of Burn and Plastic Surgery, General Hospital of Ningxia Medical University to extract human hypertrophic scar fibroblasts and human normal skin fibroblasts, respectively. Hematoxylin-eosin staining was used to identify normal skin and hypertrophic scar and immunofluorescence was used to identify fibroblasts. The relative expression level of MicroRNA-382-5p was detected by quantitative real-time PCR at the tissue level. Hypertrophic scar fibroblasts were randomly divided into shikonin group (shikonin was dissolved in dimethyl sulfone to make drug solution at a concentration of 13.46 μmol/L, which was used for cell culture for 24 hours), dimethyl sulfone group, MicroRNA-382-5p negative control group, MicroRNA-382-5p inhibitor group, shikonin+MicroRNA-382-5p negative control group and shikonin+MicroRNA-382-5p overexpression group. Real-time fluorescence quantitative PCR and western blot were used to detect the expression of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin at mRNA and protein levels, respectively. Cell counting kit-8 was used to detect cell viability. Cell scratch test was used to detect the migration ability of cells. RESULTS AND CONCLUSION: Compared with normal skin fibroblasts, hypertrophic scar fibroblasts had stronger proliferative activity (P < 0.01). Compared with the dimethyl sulfone group, shikonin down-regulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in human hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Compared with normal skin, MicroRNA-382-5p was highly expressed in hypertrophic scar (P < 0.01), and shikonin could down-regulate the expression of MicroRNA-382-5p (P < 0.01). Inhibition of MicroRNA-382-5p down-regulated the expression level of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and inhibited the migration of hypertrophic scar fibroblasts (P < 0.05). Under the influence of shikonin, overexpression of MicroRNA-382-5p upregulated the expression levels of Collagen Type I α1, Collagen Type III α1 and α-smooth muscle actin in hypertrophic scar fibroblasts (mRNA: P < 0.01, protein: P < 0.01), and promoted the migration of hypertrophic scar fibroblasts (P < 0.01). To conclude, shikonin can inhibit the fibrosis and migration of hypertrophic scar fibroblasts by down-regulating the expression of MicroRNA-382-5p. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：目前已有研究表明紫草素具有治疗增生性瘢痕的潜力,且miRNA参与增生性瘢痕的病理机制调控,猜测紫草素有可能通过影响 miRNA调控增生性瘢痕的发生发展. 目的：探讨紫草素通过影响MicroRNA-382-5p对人增生性瘢痕成纤维细胞纤维化的作用机制. 方法：收集宁夏医科大学总医院烧伤整形外科提供的增生性瘢痕组织及瘢痕旁正常皮肤组织(瘢痕旁3 cm以内),并分别分离出人增生性瘢 痕成纤维细胞和正常成纤维细胞用于后续实验.苏木精-伊红染色鉴定正常皮肤和增生性瘢痕；免疫荧光鉴定成纤维细胞；采用实时荧光 定量PCR从组织水平检测MicroRNA-382-5p相对表达水平.将增生性瘢痕成纤维细胞随机分为紫草素组(紫草素溶于二甲基亚砜配成浓度为 13.46 μmol/L的药物干预细胞24 h),二甲基亚砜组,MicroRNA-382-5p阴性对照组,MicroRNA-382-5p抑制组,紫草素+MicroRNA-382-5p阴性 对照组及紫草素+MicroRNA-382-5p过表达组.实时荧光定量PCR检测Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白mRNA表达； Western blot检测Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的蛋白表达；CCK-8法检测细胞活性；划痕实验检测细胞迁移能力. 结果与结论：①与正常皮肤成纤维细胞相比,增生性瘢痕成纤维细胞增殖活性更强(P < 0.01)；②与二甲基亚砜组相比,紫草素组可以下调增 生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并抑制增生性 瘢痕成纤维细胞迁移(P < 0.05)；③与正常皮肤相比,MicroRNA-382-5p在增生性瘢痕中呈高表达(P < 0.01),且紫草素能下调增生性瘢痕成纤维 细胞中MicroRNA-382-5p的表达(P < 0.01)；④敲低MicroRNA-382-5p能下调增生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑 肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并抑制增生性瘢痕成纤维细胞迁移(P < 0.05)；⑤过表达MicroRNA-382-5p能上调在紫 草素影响下增生性瘢痕成纤维细胞中Ⅰ型胶原蛋白α1,Ⅲ型胶原蛋白α1和α-平滑肌肌动蛋白的表达水平(mRNA：P < 0.01,蛋白：P < 0.01),并促 进增生性瘢痕成纤维细胞迁移(P < 0.01)；⑥提示紫草素可通过下调MicroRNA-382-5p的表达抑制增生性瘢痕成纤维细胞的纤维化和迁移. [ABSTRACT FROM AUTHOR]

: Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

العنوان البديل: Shikonin inhibits collagen deposition of human hypertrophic scar fibroblasts via PTEN. (English)

المؤلفون: 唐玉婷, 马芳, 贺茜, 王建军, 杨安宁, 吴凯, 焦运, 白志刚, 姜怡邓, 沈江涌

المصدر: Journal of Practical Medicine / Shiyong Yixue Zazhi; 6/10/2023, Vol. 39 Issue 11, p1382-1388, 7p

الملخص (بالإنجليزية): Objective To investigate the effect of shikonin on the collagen deposition of human hypertrophic scar fibroblasts from the perspective of PTEN. Methods Hematoxylin-eosin staining and Masson staining were used to identify hypertrophic scar tissue. Immunofluorescence was used to identify fibroblasts. The relative expression level of PTEN was detected by Western blot at the tissue level. Hypertrophic scar fibroblasts were randomly divided into dimethyl sulfone group, shikonin group, control group, si-NC group, shikonin+ si-NC group and shikonin+si-PTENC group, western blot were used to detect the expression of PTEN and collagen-related proteins (Col1, Col3 and α-SMA）at protein levels, respectively. Results Compared with normal skin, PTEN was lower expressed in hypertrophic scar (P < 0.05). shikonin was dissolved in dimethyl sulfone to make drug solution at IC50 concentration (13.46 μmol/L), which was used for human hypertrophic scar fibroblasts cell culture for 24 hours, shikonin down-regulated the expression levels of Col1, Col3 and α-SMA in human hypertrophic scar fibroblasts, while up-regulated the expression of PTEN (P < 0.05). Transfected with si-PTEN, the expressions of Col1, Col3 and α-SMA in human hypertrophic scar fibroblasts were increased compared with the si-NC group (P < 0.05). After co-transfection with shikonin + si-PTEN, the expressions of Col1, Col3 and α-SMA in human hypertrophic scar fibroblasts were increased compared with shikonin + si-NC group (P < 0.05). Conclusion Shikonin can inhibit collagen deposition in hypertrophic scar fibroblasts by up-regulating PTEN expression. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 目的 从 PTEN 角度探讨紫草素对人增生性瘢痕成纤维细胞胶原沉积的影响。方法 苏木 精-伊红染色 (HE 染色) 和马松染色 (Masson 染色) 鉴定增生性瘢痕组织, 免疫荧光鉴定人增生性瘢痕成纤 维细胞 (HSFBs); Western blot 检测人增生性瘢痕 (HS) 和同一个体正常皮肤 (NS) 组织中 PTEN 表达水平; 将 HSFBs 随机分组为：DMSO 组、shikonin 组、control 组、si-NC 组、si-PTEN 组、shikonin+si-NC 组 和 shikonin+ si-PTEN 组; Western blot 检测各组中 PTEN 和胶原相关蛋白 (Col1、Col3、α⁃SMA) 的表达。结果 与正 常皮肤组织相比, PTEN 在增生性瘢痕组织中呈低表达 (P < 0.05); 用 IC50 浓度 (13.46 μmol/L) 的紫草素干 预人增生性瘢痕成纤维细胞 24 h 后, 人增生性瘢痕成纤维细胞中 Col1、Col3 和 α-SMA 的表达降低 (P < 0.05), 而 PTEN 的表达升高 (P < 0.05); 转染 si-PTEN 后, 与 si-NC 组相比, 人增生性瘢痕成纤维细胞中 Col1, Col3 和 α⁃SMA 的表达升高 (P < 0.05); 共转染 shikonin + si-PTEN 后, 与 shikonin +si-NC 组相比, 人增生性瘢 痕成纤维细胞中 Col1、ol3 和 α⁃SMA 的表达升高 (P < 0.05)。结论 紫草素可通过上调 PTEN 的表达抑制 增生性瘢痕成纤维细胞的胶原沉积。 [ABSTRACT FROM AUTHOR]

: Copyright of Journal of Practical Medicine / Shiyong Yixue Zazhi is the property of Journal of Practical Medicine Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)