siRNA-Piezo2质粒转染BMSCs促进大鼠糖尿病足溃疡愈合  

Lentivirus mediated siRNA-Piezo2 plasmid transfection of BMSCs promotes the healing of diabetic foot ulcer via regulating vascular density in rats

作　　者：岳鹏莹 刘金侠 董晓娟 王翔 YUE Peng-ying;LIU Jin-xia;DONG Xiao-juan;WANG Xiang(Xi'an Innovation College of Yan'an University,Xi'an 710000;Department of Endocrinology,Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]延安大学西安创新学院护理学院,陕西西安710000 [2]青岛大学附属医院内分泌科,山东青岛266003

出　　处：《解剖科学进展》2024年第4期385-388,393,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81870615)。

摘　　要：目的探究siRNA-Piezo2蛋白调控骨髓间充质干细胞(BMSCs)向内皮细胞分化,促进大鼠糖尿病足溃疡愈合的作用及可能机制。方法收集糖尿病(DM)患者的血清样本,用二代测序技术进行全基因组测序。用RT-PCR检测Piezo2 mRNA的相对表达量。利用siRNA干扰技术构建siRNA-Piezo2基因沉默质粒,通过慢病毒转染BMSCs,检测BMSCs向内皮细胞的分化的效率。通过腹腔注射链脲佐菌素构建糖尿病足溃疡模型,根据是否转染siRNA-Piezo2质粒,将SD大鼠分成空白对照组、模型组、BMSCs组和siRNA干扰组。CD34染色检测溃疡部位血管密度,免疫组织化学染色检测胰腺组织β细胞数。结果伴有糖尿病足(DF)的DM患者血清中Piezo2 mRNA的相对表达量明显高于无DF的DM患者(P<0.001)。siRNA Piezo2转染BMSCs后细胞表面内皮细胞标志物CD31的表达量明显增加(P<0.05),促进BMSCs向内皮细胞分化,病足溃疡组织血管密度明显高于模型组(P<0.05),siRNA Piezo2促进糖尿病足溃疡组织血管再生。但对胰腺β细胞数量没有影响。结论siRNA Piezo2质粒转染BMSCs促进糖尿病足溃疡组织的愈合,与促进溃疡组织微血管重建相关。Objective To explore the effect of siRNA Piezo2 transfected into bone marrow mesenchymal stem cells(BMSCs)on the healing of diabetic foot ulcers in rats.Methods The whole genome of diabetes mellitus(DM)patients was sequenced by second generation sequencing.The relative expression of Piezo2 mRNA was detected by RT-PCR.Using siRNA interference technology to construct siRNA Piezo2 gene silencing plasmid,BMSCs were transfected by lentivirus to detect the differentiation efficiency of BMSCs into endothelial cells.The diabetic foot(DF)ulcer model was established by intraperitoneal injection of streptozotocin.SD rats were divided into the blank control group,the model group,the BMSCs group and siRNA interference group.CD34 staining was used to detect the density of blood vessels in ulcer area,and immunohistochemical staining was used to detect the number ofβcells in pancreas.Results The relative expression of Piezo2 mRNA was significantly higher in DM patients with DF than in DM patients without DF.siRNA Piezo2 transfected BMSCs upregulated significantly the expression level of CD31,promoting the differentiation of BMSCs into endothelial cells,increased the vascular density of diabetic foot ulcer tissue,promoting the vascular regeneration of diabetic foot ulcer tissue,with no effect on the number of pancreaticβcells.Conclusion siRNA Piezo2 plasmid can promote the differentiation of BMSCs into endothelial cells,promote the microvascular reconstruction of ulcer tissue and promote the healing of ulcer tissue.

关 键 词：糖尿病足 SIRNA 溃疡 愈合 大鼠 

分 类 号：R587.2[医药卫生—内分泌]