人唾液外泌体对大鼠皮肤损伤的修复作用  

Repair effects of human saliva-derived exosomes on cutaneous wound healing:An experimental study in rats

作　　者：宋思嘉 陈文霞[1] 李贤玉 SONG Sijia;CHEN Wenxia;LI Xianyu(College&Hospital of Stomatology,Guangxi Medical University,Guangxi Clinical Research Center for Craniofacial Deformity,Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment,Guangxi Health Commission Key Laboratory of Prevention and Treatment for Oral Infectious Diseases,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学口腔医学院,附属口腔医院,广西颅颌面畸形临床医学研究中心,颌面外科疾病诊治研究重点实验室,广西壮族自治区卫生健康委员会口腔感染性疾病防治重点实验室,广西南宁530021

出　　处：《口腔颌面外科杂志》2022年第3期152-157,共6页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81760195);广西自然科学基金(2018GXNSFAA050083)。

摘　　要：目的:探讨人唾液外泌体(saliva-derived exosomes,S-Exos)对大鼠皮肤损伤修复的影响。方法:收集健康成年人唾液,分离并鉴定唾液外泌体。将18只SD大鼠随机分为对照组、唾液处理组和外泌体处理组。通过皮肤打孔法制备大鼠皮肤全层缺损模型,各组在建模后48 h时于创面边缘分4点进行皮下注射给药100μL。在建模后第0、4、8、12、14天时观察各组大鼠创面愈合情况,计算伤口愈合率。于建模后第14和第35天处死大鼠,取创面皮肤组织,采用HE染色观察皮肤组织形态表现。结果:在透射电镜观察下,S-Exos呈杯状,粒径大小约为70 nm,蛋白质免疫印迹(Western blotting)结果显示,外泌体特异性标志物CD63和CD9均为阳性。动物实验表明,S-Exos可以促进大鼠皮肤伤口面积收缩。在建模后第8天,外泌体处理组的愈合率显著高于对照组和唾液处理组(P<0.01)。建模后第14天,外泌体处理组可见创面新生组织中成纤维细胞和炎症细胞的细胞数目及血管生成增多,建模后第35天可见胶原纤维有序平行排列,并有个别毛囊生成。结论:人唾液外泌体能够促进大鼠皮肤全层损伤的愈合,调控损伤修复过程中的炎症反应,诱导成纤维细胞的增殖,有助于损伤部位皮肤组织恢复正常的组织形态。Objective:To investigate the effect of human saliva-derived exosomes(S-Exos)on the healing of skin injury in rats.Methods:Saliva samples of healthy adults were collected and S-Exos were isolated and identified.18 Sprague Dawley(SD)rats were randomly divided into control group,saliva group and S-Exos group respectively.Full-thickness excisional skin wounds were created with a skin puncher at the dorsum of each rat,and a full-thickness dermal defect model was established.100μL ingredients of S-Exos were injected subcutaneously at four sites of the wound margin at 48 hours after model establishment.The healing situation of wounds was observed at 0,4,8,12,and 14 days after model establishment,and the wound healing rate was calculated.The rats were sacrificed at 14 and 35 days after model establishment and skin samples in defect area were harvested.HE staining was used to observe the histological morphology of skin.Results:S-Exos exhibited cup-shaped morphologies,the particle size was about 70 nm under transmission electron microscope(TEM).Western blotting results showed CD63 and CD9 as specific markers of exosomes were positively expressed in S-Exos.S-Exos could promote the contraction of rat skin wound area.At 8 days after model establishment,the wound heal ing rate of S-Exos group was significantly higher than that of the control and salivary groups(P<0.01).The skin histopathological evaluation showed that the number of fibroblasts and inflammatory cells and microvessels in S-Exos group increased at 14 days after model establishment.Collagen fibers were orderly and arranged in parallel and individual hair follicles were formed at 35 days after model establishment.Conclusion:Human S-Exos could promote wound healing of full-thickness skin defect injury in rats,modulate inflammatory process during wound healing inducing fibroblasts proliferation,and benefit healing to restore to normal histomorohology.

关 键 词：皮肤损伤 唾液 外泌体 损伤修复 

分 类 号：R782[医药卫生—口腔医学]