摘要
目的探讨人参皂苷Rg1(GRg1)通过调控丙酮酸激酶M2(PKM2)的表达对人视网膜微血管内皮细胞(HRMECs)糖酵解的影响。方法在体外培养HRMECs,将其分为对照组(NC)组、高葡萄糖(HG)组、HG+人参皂苷Rg1(HG+GRg1)组、HG+人参皂苷Rg1+低表达PKM2(HG+GRg1+si-PKM2)组、HG+人参皂苷Rg1+过表达PKM2(HG+GRg1+OE-PKM2)组,si-PKM2、OE-PKM2通过细胞转染方式转染至HRMECs中。采用qRT-PCR法检测HRMECs中PKM2 mRNA的表达情况;通过Western blot检测HRMECs中相关蛋白表达量;在倒置显微镜下观察细胞体外管腔生成数量,以量化血管形成能力;收集各组细胞培养液,分别用葡萄糖检测试剂盒、乳酸检测试剂盒和腺嘌呤核苷三磷酸(ATP)检测试剂盒检测葡萄糖摄取量、乳酸产量以及ATP含量。结果HG诱导会显著增加HRMECs的血管形成数量、糖酵解及PKM2的表达,而在加入GRg1处理后,HG所致的血管数量、糖酵解及PKM2的表达均明显减少;转染si-PKM2可协助GRg1对糖酵解和血管形成的抑制作用,而转染OE-PKM2会干扰GRg1的功能。结论GRg1通过抑制PKM2减少HRMECs糖酵解抑制血管形成。
Objective To investigate the effect of ginsenoside Rg1(GRg1)on human retinal microvasculaRendothelial cells(HRMECs)glycolysis by regulating pyruvate kinase M2(PKM2)expression.Methods HRMECs were cultured in vitro and divided into normal control(NC)group,high glucose(HG)group,high glucose+ginsenoside Rg1(HG+GRg1)group,high glucose+ginsenoside Rg1+low expression PKM2(HG+GRg1+si-PKM2)group,and high glucose+ginsenoside Rg1+overexpression PKM2(HG+GRg1+OE-PKM2)group.si-PKM2 and OE-PKM2 were transfected into HRMECs cells by cell transfection.The expression of PKM2 mRNA in HRMECs was detected by qRT-PCR.The expression levels of related proteins in HRMECs were detected by Western blot.The numbeRof lumen formation in vitro was observed undeRan inverted microscope to quantify the angiogenesis ability.Cell culture medium of each group was collected,and glucose intake,lactate production and adenosine triphosphate(ATP)content were detected by glucose detection kit,lactate detection kit and ATP detection kit,respectively.Results HG induced HRMECs significantly increased the numbeRof blood vessel formation,glycolysis and PKM2 expression,while GRg1 treatment significantly reduced the numbeRof blood vessel formation,glycolysis and PKM2 expression;transfection of si-PKM2 assisted the inhibitory effect of GRg1 on glycolysis and angiogenesis while transfection of OE-PKM2 interfered with the function of GRg1.Conclusion GRg1 inhibits angiogenesis by inhibiting PKM2 to reduce glycolysis of HRMECs.
作者
薛黎萍
胡敏
李亚娣
张晓帆
张洁莹
周园
梁佳芮
张传宏
丁鹏
Xue Liping;Hu Min;Li Yadi;Zhang Xiaofan;Zhang Jieying;Zhou Yuan;Liang Jiarui;Zhang Chuanhong;Ding Peng(Dept of Pediatric Ophthalmology,Affiliated Hospital of Yunnan University,Kunming 650000;Dept of Neurosurgery,The First Affiliated Hospital of Kunming Medical University,Kunming 650000)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第10期1559-1564,共6页
Acta Universitatis Medicinalis Anhui
基金
云南省中青年学术和技术带头人后备人才项目(编号:202005AC160021)
云南省科技厅科技计划项目[编号:2019FE001(-169)]。
