The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, P.R. China.
Jiangxi Health Vocational College, Nanchang, China.
Int J Low Extrem Wounds. 2023 Dec;22(4):704-712. doi: 10.1177/15347346211038092. Epub 2021 Aug 30.
To investigate the effect of MicroRNA-146a modified adipose-derived stem cell exosomes on the proliferation and migration of fibroblasts and the therapeutic effect on wound healing. Culture and identification of human adipose-derived stem cells (hASCs), miRNA-146a minic vector was constructed and transfected into hASCs, the exosomes of the empty group and overexpression group were extracted, identified, and quantitatively analyzed after 24 h of successful transfection. The exosomes were added into National Institute of Health Mouse Embryonic Fibroblasts (NIH/3T3) and cultured for 48 h, the proliferation and migration ability of NIH/3T3 fibroblasts was detected. The expression of serpin family H member 1 (SERPINH1) and phosphorylated extracellular regulated protein kinase (p-ERK) was detected by Western blot. The model of back wound was established. The exosomes were injected into 4 different sites with the shape of "cross" around the wound, and the scar diameter of the skin defect was measured at 3, 7, and 11 days, the skin of the defect was taken on the 14th day. platelet endothelial cell adhesion molecule-1 (CD31) was detected by immunofluorescence staining to evaluate angiogenesis, and Western blot was used to detect the expression of SERPINH1 and p-ERK. The miR-146a mimic-exosome promoted the proliferation and migration of fibroblasts, and the expression of SERPINH1 and p-ERK2 was up-regulated. After the rats were treated with exosomes, the wound area decreased rapidly, neovascularization was promoted, and the expression of SERPINH1 and p-ERK2 was up-regulated. MicroRNA-146a modified adipose stem cell exosomes could regulate the expression of SERPINH1 and p-ERK, promote the migration and proliferation of fibroblasts, and neovascularization to promote the wound healing of rat back.
探讨微小 RNA-146a 修饰的脂肪来源干细胞外泌体对成纤维细胞增殖和迁移的影响及其对伤口愈合的治疗作用。
培养并鉴定人脂肪来源干细胞（hASCs），构建 miRNA-146a 微小载体并转染 hASCs，转染 24 h 后提取空载体组和过表达组的外泌体，进行鉴定和定量分析。将外泌体加入 NIH 小鼠胚胎成纤维细胞（NIH/3T3）中培养 48 h，检测 NIH/3T3 成纤维细胞的增殖和迁移能力。Western blot 检测丝氨酸蛋白酶抑制剂家族 H 成员 1（SERPINH1）和磷酸化细胞外调节蛋白激酶（p-ERK）的表达。建立背部伤口模型，在伤口周围以“十字”形注射 4 个不同部位的外泌体，测量第 3、7、11 天皮肤缺损的瘢痕直径，第 14 天取皮肤缺损部位。免疫荧光染色检测血小板内皮细胞黏附分子-1（CD31），评估血管生成，Western blot 检测 SERPINH1 和 p-ERK 的表达。
miR-146a 模拟物-外泌体促进成纤维细胞的增殖和迁移，上调 SERPINH1 和 p-ERK2 的表达。外泌体处理大鼠后，伤口面积迅速减小，促进新生血管形成，上调 SERPINH1 和 p-ERK2 的表达。
miR-146a 修饰的脂肪干细胞外泌体可调节 SERPINH1 和 p-ERK 的表达，促进成纤维细胞的迁移和增殖，促进新生血管形成，从而促进大鼠背部伤口愈合。
