Chen Liushan, Qin Linghao, Chen Chujun, Hu Qiong, Wang Junjian, Shen Juan
Department of Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangdong Pharmaceutical University, Guangzhou, Guangdong, China.
School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou, Guangdong, China.
Cell Biol Int. 2021 Sep;45(9):1976-1985. doi: 10.1002/cbin.11627. Epub 2021 Jun 3.
Nonhealing wounds in diabetes remain a global clinical and research challenge. Exosomes are primary mediators of cell paracrine action, which are shown to promote tissue repair and regeneration. In this study, we investigated the effects of serum derived exosomes (Serum-Exos) on diabetic wound healing and its possible mechanisms. Serum-Exos were isolated from blood serum of normal healthy mice and identified by transmission electron microscopy and western blot. The effects of Serum-Exos on diabetic wound healing, fibroblast growth and migration, angiogenesis and extracellular matrix (ECM) formation were investigated. Our results showed that the isolated Serum-Exos exhibited a sphere-shaped morphology with a mean diameter at 150 nm, and expressed classical markers of exosomes including HSP70, TSG101, and CD63. Treatment with Serum-Exos elevated the percentage of wound closure and shortened the time of healing in diabetic mice. Mechanistically, Serum-Exos promoted granulation tissue formation and increased the expression of CD31, fibronectin and collagen-ɑ in diabetic mice. Serum-Exos also promoted the migration of NIH/3T3 cells, which was associated with increased expression levels of PCNA, Ki67, collagen-α and fibronectin. In addition, Serum-Exos enhanced tube formation in human umbilical vein endothelial cells and induced the expression of CD31 at both protein and messenger RNA levels. Collectively, our results suggest that Serum-Exos may facilitate the wound healing in diabetic mice by promoting angiogenesis and ECM formation, and show the potential application in treating diabetic wounds.
糖尿病患者的难愈合伤口仍然是全球临床和研究面临的挑战。外泌体是细胞旁分泌作用的主要介质，已被证明可促进组织修复和再生。在本研究中，我们调查了血清来源的外泌体（血清外泌体）对糖尿病伤口愈合的影响及其可能的机制。从正常健康小鼠的血清中分离出血清外泌体，并通过透射电子显微镜和蛋白质印迹法进行鉴定。研究了血清外泌体对糖尿病伤口愈合、成纤维细胞生长和迁移、血管生成以及细胞外基质（ECM）形成的影响。我们的结果表明，分离出的血清外泌体呈现球形形态，平均直径为150 nm，并表达外泌体的经典标志物，包括热休克蛋白70（HSP70）、肿瘤易感基因101（TSG101）和CD63。血清外泌体治疗可提高糖尿病小鼠伤口闭合的百分比并缩短愈合时间。从机制上讲，血清外泌体促进肉芽组织形成，并增加糖尿病小鼠中CD31、纤连蛋白和胶原蛋白α的表达。血清外泌体还促进NIH/3T3细胞的迁移，这与增殖细胞核抗原（PCNA）、Ki67、胶原蛋白α和纤连蛋白表达水平的增加有关。此外，血清外泌体增强人脐静脉内皮细胞的管腔形成，并在蛋白质和信使RNA水平上诱导CD31的表达。总体而言，我们的结果表明，血清外泌体可能通过促进血管生成和ECM形成来促进糖尿病小鼠的伤口愈合，并显示出在治疗糖尿病伤口方面的潜在应用价值。
