Effect of MicroRNA-146a Modified Adipose-Derived Stem Cell Exosomes on Rat Back Wound Healing.

IF 1.5 4区 医学 Q3 DERMATOLOGY
Gangquan Chen Md, Yaling Wu Md, Lijin Zou Md, Yuanlin Zeng Md
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引用次数: 13

Abstract

Objective: 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. Methods: 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. Results: 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. Conclusions: 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.

MicroRNA-146a修饰脂肪源性干细胞外泌体对大鼠背部伤口愈合的影响
目的:研究MicroRNA-146a修饰的脂肪干细胞外泌体对成纤维细胞增殖和迁移的影响以及对伤口愈合的治疗作用。方法:培养和鉴定人脂肪源性干细胞,构建miRNA-146a minic载体并转染hASCs,提取空组和过表达组的外泌体,鉴定并在24小时后定量分析 h成功转染。将外泌体加入美国国立健康研究所小鼠胚胎成纤维细胞(NIH/3T3)中并培养48 h、 检测NIH/3T3成纤维细胞的增殖和迁移能力。用蛋白质印迹法检测血清蛋白家族H成员1(SERPINH1)和磷酸化细胞外调节蛋白激酶(p-ERK)的表达。建立背部创伤模型。将外泌体注射到伤口周围4个形状为“十字架”的不同部位,并在第3、7和11天测量皮肤缺损的疤痕直径,在第14天取下缺损的皮肤。免疫荧光染色检测血小板内皮细胞粘附分子-1(CD31)以评价血管生成,Western印迹检测SERPINH1和p-ERK的表达。结果:miR-146a模拟外泌体促进成纤维细胞的增殖和迁移,SERPINH1和p-ERK2的表达上调。用外泌体处理大鼠后,伤口面积迅速减少,促进新生血管形成,SERPINH1和p-ERK2的表达上调。结论:MicroRNA-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,促进大鼠背部伤口愈合。
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来源期刊
CiteScore
4.60
自引率
17.60%
发文量
95
审稿时长
>12 weeks
期刊介绍: The International Journal of Lower Extremity Wounds (IJLEW) is a quarterly, peer-reviewed journal publishing original research, reviews of evidence-based diagnostic techniques and methods, disease and patient management, and surgical and medical therapeutics for lower extremity wounds such as burns, stomas, ulcers, fistulas, and traumatic wounds. IJLEW also offers evaluations of assessment and monitoring tools, dressings, gels, cleansers, pressure management, footwear/orthotics, casting, and bioengineered skin. This journal is a member of the Committee on Publication Ethics (COPE).
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