التفاصيل البيبلوغرافية
| العنوان: |
miR-382-3p 抑制人增生性瘢痕成纤维细胞的增殖. (Chinese) |
| العنوان البديل: |
miR-382-3p inhibits the proliferation of human hypertrophic scar fibroblasts. (English) |
| المؤلفون: |
贺 茜, 马 芳, 万 瑀, 唐玉婷, 马胜超, 姜怡邓, 沈江涌 |
| المصدر: |
Chinese Journal of Tissue Engineering Research / Zhongguo Zuzhi Gongcheng Yanjiu; 4/18/2023, Vol. 27 Issue 11, p1758-1764, 7p |
| مصطلحات موضوعية: |
PROLIFERATING cell nuclear antigen, HYPERTROPHIC scars, CYCLIN-dependent kinase inhibitors, STAT proteins, GENE expression, FIBROBLASTS, CYCLIN-dependent kinases |
| الملخص (بالإنجليزية): |
BACKGROUND: At present, numerous studies have shown that miRNA is involved in the occurrence and development of hypertrophic scars and STAT1 is involved in the proliferation of scar fibroblasts. It is speculated that miR-382-3p may also be related to the occurrence and development of hypertrophic scars. OBJECTIVE: To investigate the mechanism of miR-382-3p on the proliferation of human hypertrophic scar fibroblasts. METHODS: Hypertrophic scar and normal skin of the same individual were collected from the Department of Burn Plastic Surgery, General Hospital of Ningxia Medical University, and human hypertrophic scar fibroblasts and human normal skin fibroblasts were then extracted. Cell transfection was conducted as follows: (1) the cells were divided into control group (no treatment), miR-382-3p negative control group, and miR-382-3p overexpression group; (2) the cells were divided into control group (no treatment), STAT1 interference control group (si-NC) and STAT1 interference groups (si-STAT1-1, si-STAT1-2, si-STAT1-3). Hematoxylin-eosin staining was used to identify normal skin and hypertrophic scar, and immunofluorescence was used to identify fibroblasts. Quantitative realtime PCR was applied to detect the mRNA expression of miR-382-3p, STAT1, proliferating cell nuclear antigen (PCNA) and cyclin-dependent kinase inhibitor (p27). Western blot assay was performed to detect the protein expressions of STAT1, PCNA and p27. Cell counting kit-8 and EdU were used to detect cell proliferation activity and proliferation level. Targetscan was used to predict the downstream target genes of miR-382-3p and dual luciferase was used to verify the binding of miR-382-3p to STAT1. RESULTS AND CONCLUSION: Compared with normal skin and normal skin fibroblasts, miR-382-3p was lowly expressed in hypertrophic scar and hypertrophic scar fibroblasts (tissue: P < 0.01, cell: P < 0.01), and STAT1 was highly expressed in hypertrophic scar and hypertrophic scar fibroblasts (mRNA level in tissue: P < 0.01, protein level in tissue: P < 0.01; mRNA level in cells: P < 0.01, protein level in tissue: P < 0.01). After overexpression of miR-382-3p, the cell proliferation ability was weakened (P < 0.05), the number of EdU positive cells was decreased (P < 0.01), the expression of PCNA was decreased (mRNA: P < 0.01, protein: P < 0.05), and the expression of p27 was increased (mRNA: P < 0.05, protein: P < 0.05). miR-382-3p could target and regulate the expression of STAT1 (P < 0.01), and overexpression of miR-382-3p could reduce the expression of STAT1 (mRNA: P < 0.01, protein: P < 0.01). Interference with STAT1 reduced the expression of PCNA (P < 0.05), increased the expression of p27 (P < 0.05), and reduced the number of EdU positive cells (P < 0.01). To conclude, miR-382-3p could inhibit the proliferation of hypertrophic scar fibroblasts by inhibiting the expression of STAT1, which provides a certain theoretical basis for identifying effective targets for the treatment of hypertrophic scars. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
背景：目前多项研究证实了miRNA影响增生性瘢痕的发生发展，STAT1参与瘢痕成纤维细胞的增殖过程，猜测miR-382-3p也有可能与增生性 瘢痕的发生发展有关系。目的：探讨miR-382-3p对人增生性瘢痕成纤维细胞增殖的作用机制。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和同一个体正常皮肤，提取人增生性瘢痕成纤维细胞和人正常皮肤成纤 维细胞；细胞转染分别设置：①对照组(不做处理)、miR-382-3p阴性对照组、miR-382-3p过表达组；②对照组(不做处理)、STAT1干扰对照 组(si-NC)和STAT1干扰组(si-STAT1-1、si-STAT1-2、si-STAT1-3)。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； qRT-PCR检测miR-382-3p、STAT1、增殖细胞核抗原及细胞周期依赖性激酶抑制物(p27)mRNA表达；Western blot检测STAT1、增殖细胞核抗原 及p27蛋白表达；CCK8、EdU检测细胞增殖活力和增殖水平；Targetscan预测miR-382-3p的下游靶基因，双荧光素酶验证miR-382-3p与STAT1 的结合情况。 结果与结论：①与正常皮肤和正常皮肤成纤维细胞相比，miR-382-3p在增生性瘢痕和增生性瘢痕成纤维细胞中呈低表达(组织：P < 0.01，细胞：P < 0.01)，STAT1在增生性瘢痕和增生性瘢痕成纤维细胞中呈高表达(组织水平mRNA：P < 0.01，组织水平蛋白：P < 0.01；细胞水平 mRNA：P < 0.01，细胞水平蛋白：P < 0.01)；②过表达miR-382-3p后细胞增殖能力减弱(P < 0.05)，EdU阳性细胞数减少(P < 0.01)，增殖细胞 核抗原的表达减少(mRNA：P < 0.01，蛋白：P < 0.05)，p27的表达增加(mRNA：P < 0.05，蛋白：P < 0.05)；③miR-382-3p能够靶向调控STAT1 的表达(P < 0.01)，过表达miR-382-3p可导致STAT1的表达减少(mRNA：P < 0.01，蛋白：P < 0.01)；④干扰STAT1后可导致增殖细胞核抗原的表 达减少(P < 0.05)，p27的表达增加(P < 0.05)，EdU阳性细胞数减少(P < 0.01)；⑤结果表明：miR-382-3p可通过抑制STAT1的表达进而抑制增生 性瘢痕成纤维细胞的增殖，为未来增生性瘢痕的治疗寻找有效靶点提供一定的理论依据。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
