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: Erastin inhibits proliferation of hypertrophic scar fibroblasts. (English)

المؤلفون: 贺 茜, 万 瑀, 唐玉婷, 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; 11/18/2023, Vol. 27 Issue 32, p5120-5125, 6p

مصطلحات موضوعية: PROLIFERATING cell nuclear antigen, HYPERTROPHIC scars, CYCLIN-dependent kinase inhibitors, FERRITIN, GENE expression, IRON ions

الملخص (بالإنجليزية): BACKGROUND: Hypertrophic scar is a kind of pathological scar that appears in the healing process after skin trauma caused by various reasons and there is no effective treatment. OBJECTIVE: To investigate the effect of ferroptosis inducer (Erastin) on the proliferation of human hypertrophic scar fibroblasts. METHODS: Hypertrophic scar samples provided by the Burn Plastic Surgery Department of the General Hospital of Ningxia Medical University and normal skin samples of the same individual were collected. Human hypertrophic scar fibroblasts were then extracted for subsequent experiments. (1) The cells were divided into control group (without treatment) and ferroptosis inducer group (treated with 20 μmol/L Erastin for 24 hours). The expression of Ferritin was detected by western blot. Iron ion detection kit was used to measure cellular iron ion concentration. Malondialdehyde detection kit was used to detect cellular malondialdehyde content. (2) The cells were divided into control group (without treatment), ferroptosis inducer group (treated with 20 μmol/L Erastin for 24 hours) and ferroptosis inducer+ferroptosis inhibitor group (co-treated with 20 μmol/L Erastin and 20 μmol/L Ferrostatin-1 for 24 hours). The mRNA and protein expressions of proliferating cell nuclear antigen (PCNA) and cyclin-dependent kinase inhibitor (p27) were detected using qRT-PCR and western blot. Cell counting kit-8 and EdU were used to detect cell proliferation viability and levels. RESULTS AND CONCLUSION: Compared with the control group, Erastin decreased the ferritin expression (P < 0.01), increased the content of iron ions (P < 0.05), and elevated the malondialdehyde content in the cells (P < 0.01). Compared with the control group, Erastin decreased the expression of PCNA (P < 0.01), increased the expression of p27 (P < 0.05), weakened the cell proliferation ability (P < 0.01), and reduced the number of EdU-positive cells (P < 0.01). Compared with the ferroptosis inducer group, the ferroptosis inducer + ferroptosis inhibitor group had increased expression of PCNA (P < 0.05), decreased p27 expression (P < 0.05), enhanced cell proliferation (P < 0.01), and increased number of EdU-positive cells (P < 0.05). To conclude, the ferroptosis inducer (Erastin) induces ferroptosis in hypertrophic scar fibroblasts and then inhibits their proliferation. [ABSTRACT FROM AUTHOR]

Abstract (Chinese): 背景：增生性瘢痕是各种原因导致的皮肤创伤后愈合过程中出现的一种病理性瘢痕，目前缺乏特效治疗方法。 目的：探讨铁死亡诱导剂Erastin对人增生性瘢痕成纤维细胞增殖的影响。 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和同一个体正常皮肤，提取人增生性瘢痕成纤维细胞进行后续实验。 将细胞分为对照组(不做处理)和铁死亡诱导剂组(用20 μmol/L的Erastin干预细胞24 h)，Western blot检测铁蛋白(Ferritin)的表达，铁离子检 测试剂盒测定细胞铁离子浓度，丙二醛检测试剂盒检测细胞丙二醛水平；将细胞分为对照组(不做处理)、铁死亡诱导剂组(用20 μmol/L的 Erastin干预细胞24 h)和铁死亡诱导剂+铁死亡抑制剂组(用20 μmol/L的Erastin和20 μmol/L的Ferrostatin-1同时干预细胞24 h)，qRT-PCR检测 PCNA及p27的mRNA表达，Western blot检测PCNA及p27的蛋白表达，CCK-8、EdU检测细胞增殖活力和增殖水平。 结果与结论：①与对照组相比，铁死亡诱导剂组铁蛋白减少(P < 0.01)，铁离子增多(P < 0.05)，丙二醛增多(P < 0.01)；②与对照组相比，铁 死亡诱导剂组PCNA的表达降低(P < 0.01)，p27的表达增加(P < 0.05)，细胞增殖能力减弱(P < 0.01)，EdU阳性细胞数减少(P < 0.01)；③与铁死 亡诱导剂组相比，铁死亡诱导剂+铁死亡抑制剂组PCNA的表达增加(P < 0.05)，p27的表达降低(P < 0.05)，细胞增殖能力增强(P < 0.01)，EdU 阳性细胞数增多(P < 0.05)；④结果表明，铁死亡诱导剂Erastin通过诱导增生性瘢痕成纤维细胞发生铁死亡进而抑制其增殖。 [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.)