高糖、低氧对大鼠心肌微血管内皮细胞新生功能影响的体外研究  

Effects of high glucose and hypoxia on the neogenesis function of rat myocardial microvascular endothelial cells in vitro

作　　者：蔡梅峰 崔永生[1] 何文凯[1] 李明琰[1] 应虹柳 曾国鹏 CAI Meifeng;CUI Yongsheng;HE Wenkai;LI Mingyan;YING Hongliu;ZENG Guopeng(Department of Cardiology, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 511400, China)

机构地区：[1]广州医科大学附属第二医院心血管内科,广东广州511400

出　　处：《暨南大学学报（自然科学与医学版）》2019年第2期109-116,共8页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金资助项目(81300086);广东省自然科学基金资助项目(2015A030313451)

摘　　要：目的:探讨高糖、低氧对大鼠心肌微血管内皮细胞(CMEC)新生功能的影响.方法:体外培养成年大鼠CMEC,随机分为4组:正常对照组(NG组)、高糖组(HG组)、低氧组(HNG组)和高糖+低氧联合组(HHG组).Real-time PCR法检测各组CMEC的HIF-1α、VEGF的mRNA表达水平; CCK-8法检测各组CMEC的体外增殖能力;transwell小室实验评价各组CMEC的迁移能力;体外小管形成实验评价各组CMEC的成管能力.结果:(1) HIF-1α的mRNA表达水平:NG组与HG组之间、HNG组与HHG组之间无统计学差异(P> 0. 05); HNG组明显高于NG组(P <0. 05),HHG组明显高于HG组(P <0. 05),表明低氧可促进CMEC的HIF-1αmRNA表达. VEGF的mRNA表达水平:NG组与HG组间无统计学差异(P> 0. 05),HNG组明显高于HHG组(P <0. 05); HNG组明显高于NG组(P <0. 05),HHG组明显高于HG组(P <0. 05),表明高糖可抑制CMEC的VEGF mRNA表达,而低氧可促进其表达.(2)细胞的增殖能力:NG组明显高于HG组(P <0. 05),HNG组明显高于HHG组(P <0. 05); NG组明显高于HNG组(P <0. 05),HG组明显高于HHG组(P <0. 05),表明高糖、低氧均可抑制CMEC增殖.(3)细胞的迁移能力:NG组明显高于HG组(P <0. 05),HNG组明显高于HHG组(P <0. 05); NG组明显高于HNG组(P <0. 05),HG组明显高于HHG组(P <0. 05),表明高糖、低氧均可抑制CMEC迁移.(4)细胞的成管能力:NG组明显高于HG组(P <0. 05),HNG组明显高于HHG组(P <0. 05); NG组明显高于HNG组(P <0. 05),HG组明显高于HHG组(P <0. 05),表明高糖、低氧均可抑制CMEC小管形成能力.结论:高糖对CMEC的HIF-1αmRNA表达的影响不明显,但可明显抑制CMEC的VEGF mRNA表达;低氧可促进CMEC的HIF-1α和VEGF mRNA表达;高糖和低氧均可降低CMEC的增殖能力、迁移能力和成管能力,两者联合时作用更为明显.Objective: To investigate the effects of high glucose and hypoxia on the neogenesis function of rat myocardial microvascular endothelial cells( CMEC) in vitro. Methods: Adult rat CMEC were cultured in vitro and randomly divided into four groups: normal control group( NG group),high glucose group( HG group),hypoxia group( HNG group),and high glucose and hypoxia group( HHG group). Real-time PCR assay was used for detecting the mRNA level of HIF-1α and VEGF. CCK-8 assay was used for detecting the proliferation ability of CMEC. Transwell assay was used for evaluating migration ability of CMEC,and capillary-like tube formation assay was used for evaluating tube formation ability of CMEC. Results: As for the expression level of HIF-1α mRNA,there was no significant difference between NG group and HG group,HNG group and HHG group( P > 0. 05);the expression level was significantly higher in HNG group than that in NG group( P < 0. 05) and HHG group significantly higher than HG group( P < 0. 05). It indicated that hypoxia can promote the expression of HIF-1α mRNA in CMEC. As for the expression level of VEGF mRNA,there was no significant difference between NG group and HG group( P > 0. 05);the expression level in HNG group was significantly higher than that in HHG group( P < 0. 05),HNG group significantly higher than NG group( P < 0. 05) and HHG group significantly higher than HG group( P < 0. 05). It indicated that high glucose can inhibit the expression of VEGF mRNA in CMEC,while hypoxia can promote it. The proliferation,cell migration capacity and tube forming ability of the cells in NG group was significantly higher than that in HG group( P < 0. 05),HNG group significantly higher than HHG group( P <0. 05),NG group significantly higher than HNG group( P < 0. 05),and HG group significantly higher than HHG group( P < 0. 05). It indicated that both high glucose and hypoxia can inhibit the proliferation,migration and tube forming ability of CMEC. Conclusion: High glucose has no significant effect on the expression of HIF-1

关 键 词：低氧诱导因子1Α 心肌微血管内皮细胞 高葡萄糖 低氧 

分 类 号：R331.32[医药卫生—人体生理学]