Nr-CWS增强ADSC分泌功能提高糖尿病创面治疗效果的研究  

Effects of Nr-CWS on the treatment of diabetic wounds by enhancing the secretion function of ADSCs

作　　者：郭磊 刘畅 张化彬 李强 金培生 GUO Lei;LIU Chang;ZHANG Huabing;LI Qiang;JIN Peisheng(Department of Plastic Surgery,the Affiliated Hospital of Xuzhou Medical University,Xuzhou,Jiangsu 221002,China)

机构地区：[1]徐州医科大学附属医院整形外科,江苏徐州221002

出　　处：《徐州医科大学学报》2021年第11期810-816,共7页Journal of Xuzhou Medical University

基　　金：中国博士后科学基金(2019T120462);江苏省博士后科学基金(2019K155)。

摘　　要：目的探讨红色诺卡菌细胞壁骨架(Nocardia rubra cell wall skeleton,Nr-CWS)通过增强脂肪来源干细胞(adipose derived stem cell,ADSC)的分泌功能提高糖尿病创面的治疗效果。方法离体制备ADSC细胞并鉴定。制备ADSC细胞上清液(conditioned medium of ADSC,ADSC-CM)和用Nr-CWS刺激ADSC的细胞上清液(ADSC-CM^(Nr-CWS))。将人脐静脉内皮细胞(HUVEC)分为3组:高糖对照组、ADSC-CM组、ADSC-CM^(Nr-CWS)组。ELISA法检测ADSC-CM的细胞因子含量。采用CCK-8法、Transwell实验、划痕实验、血管形成实验检测ADSC-CM^(Nr-CWS)对于HUVEC增殖、侵袭、迁徙和血管生成能力的影响。在体实验中,建立小鼠糖尿病创面动物模型,将小鼠分为高糖对照组、ADSC-CM组、ADSC-CM^(Nr-CWS)组,每组10只。将ADSC-CM^(Nr-CWS)注射于创面周缘,分析创面愈合速度和质量,以及创面血管新生水平。结果离体实验中,与高糖对照组、ADSC-CM组相比,ADSC-CM^(Nr-CWS)组含有更多的血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)、肝细胞生长因子(HGF)(P<0.05);ADSC-CM^(Nr-CWS)组HUVEC的增殖、侵袭、迁徙及血管形成作用显著增强,差异有统计学意义(P<0.05)。在体实验中,与高糖对照组、ADSC-CM组相比,ADSC-CM^(Nr-CWS)组实验动物创面的血管新生水平及创面愈合率均显著上升,创面修复质量也有显著改善,差异有统计学意义(P<0.05)。结论Nr-CWS可通过提高ADSC分泌功能,促进糖尿病创面的血管新生来及创面愈合。Objective To explore the effects of Nocardia rubra cell wall skeleton(Nr-CWS)on diabetic wounds by enhancing the secretion function of adipose derived stem cells(ADSCs).Methods ADSCs were prepared in vitro and identified.Then,the conditional medium of ADSCs(ADSC-CM)and those stimulated by Nr-CWS(ADSC-CM^(Nr-CWS)) were prepared.Human umbilical vein endothelial cells(HUVECs)were divided into three groups:a high-glucose control group,an ADSC-CM group and an ADSC-CM^(Nr-CWS) group.The contents of cytokines in ADSC-CM were detected by ELISA.The proliferation,invasion,migration and angiogenesis of HUVECs were measured by CCK-8 assay,Transwell assay,wound healing test and angiogenesis test,respectively.For in vivo experiments,a diabetic wound model of mice was established.ADSC-CM^(Nr-CWS) were injected around the wound surface,and the speed and quality of wound healing and wound angiogenesis were analyzed.Results Compared with the high-glucose control group and ADSC-CM group,the ADSC-CM^(Nr-CWS) group released increased amounts of vascular endothelial growth factor(VEGF),basic fibroblast growth factor(bFGF)and hematopoietic growth factor(HGF)(P<0.05),and HUVECs in the ADSC-CM^(Nr-CWS) group presented remarkable improvement in the proliferation,invasion,migration and angiogenesis(P<0.05).For in vivo studies,compared with the high-glucose control group and ADSC-CM group,the ADSC-CM^(Nr-CWS) group showed obvious increases in wound angiogenesis and wound healing rate,and the quality of wound repair was also significantly improved(P<0.05).Conclusions Nr-CWS can promote the angiogenesis of diabetic wounds and wound healing by improving the secretion function of ADSCs.

关 键 词：红色诺卡菌细胞壁骨架 脂肪来源干细胞 糖尿病创面 血管新生 

分 类 号：R62[医药卫生—整形外科]