摘要
目的 探讨达格列净(DAPA)对高糖环境下体外培养的大鼠内皮祖细胞(EPCs)功能的影响。方法 采用荧光染色法对SD大鼠骨髓来源EPCs进行鉴定。以EPCs为研究对象,设立对照组(CG组)、高糖组(HG组)、高糖+DAPA组(GD组)、高糖+DAPA+LY294002组(GDL组)。分别采用MTT法、流式细胞术、小管形成实验检测EPCs细胞活力、凋亡、形成小管能力。应用Western blot检测丝氨酸/苏氨酸蛋白激酶(AKT)/内皮型一氧化氮合酶(eNOS)信号通路相关蛋白表达。结果 与CG组相比较,HG组EPCs细胞活力、形成小管能力、磷酸化AKT(p-AKT)和磷酸化eNOS(p-eNOS)的蛋白表达以及p-AKT/AKT和p-eNOS/eNOS比值均降低(P<0.05),而EPCs的凋亡率增加(P<0.05);与HG组相比较,GD组EPCs的细胞活力、形成小管能力、p-AKT和p-eNOS的蛋白表达以及p-AKT/AKT和p-eNOS/eNOS比值均增加(P<0.05),而EPCs的凋亡率降低(P<0.05);与GD组相比较,GDL组EPCs的细胞活力、形成小管能力、p-AKT和p-eNOS的蛋白表达以及p-AKT/AKT和p-eNOS/eNOS比值均降低(P<0.05),而EPCs的凋亡率增加(P<0.05)。结论 DAPA可通过AKT/eNOS途径保护EPCs免受高糖诱导的功能损害。
Objective To explore the effect of dapagliflozin(DAPA)on the function of rat endothelial progenitor cells(EPCs)cultured in vitro in a high glucose environment.Methods Bone marrow derived EPCs from sprague-dawley(SD)rats were identified by fluorescence staining.EPCs were divided into control group(CG group),high glucose group(HG group),high glucose+DAPA group(GD group)and high glucose+DAPA+LY294002 group(GDL group).MTT assay,flow cytometry,tubule formation assay were used to detect the viability,apoptosis,tubule formation ability of EPCs,respectively.Western blot was used to detect the protein expression of AKT/eNOS signaling pathway.Results Compared with CG group,cell viability,the ability to form tubules,the protein expression of p-AKT and p-eNOS,and the ratio of p-AKT/AKT and p-eNOS/eNOS in HG group significantly decreased(P<0.05),while the apoptosis rate of EPCs significantly increased(P<0.05).Compared with HG group,cell viability,the ability to form tubules,the protein expression of p-AKT and p-eNOS,and the ratio of p-AKT/AKT and p-eNOS/eNOS in GD group significantly increased(P<0.05),while the apoptosis rate of EPCs was significantly reduced(P<0.05).Compared with GD group,cell viability,the ability to form tubules,the protein expression of p-AKT and p-eNOS,and the ratio of p-AKT/AKT and p-eNOS/eNOS in GDL group significantly decreased(P<0.05),while the apoptosis rate of EPCs significantly increased(P<0.05).Conclusion DAPA can protect EPCs from high glucose induced functional damage through AKT/eNOS pathway.
作者
解丹丹
巫婷婷
赵晓彤
许慕蓉
陈明卫
Xie Dandan;Wu Tingting;Zhao Xiaotong;Xu Murong;Chen Mingwei(Dept of Endocrinology, The First Affiliated Hospital of Anhui Medical University, Hefei 230022)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第6期957-962,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省重点研究与开发计划项目(编号:202004a0702-0016)
安徽省自然科学基金(编号:2108085MH269)。
关键词
达格列净
糖尿病
内皮祖细胞
AKT
ENOS
dapagliflozin
diabetes mellitus
endothelial progenitor cell
AKT
eNOS
