摘要
目的探讨雌二醇(E2)与同型半胱氨酸(Hcy)对人脐静脉内皮细胞(HUVEC)增殖和凋亡的影响及可能的作用机制。方法用不同浓度的E2和Hcy处理HUVEC,应用CCK-8法检测细胞的增殖能力,流式细胞术AnnexinⅤ-FITC/PI双染检测细胞的凋亡率。选取最适浓度的E2和Hcy,设对照组、E2组、Hcy组和E2+Hcy组。采用实时定量RCR和Western blotting检测细胞中MTHFR mRNA和蛋白的表达水平。结果CCK-8法结果显示,E2对HUVEC有一定的促增殖作用,Hcy对HUVEC有抑制增殖的作用。流式细胞术结果显示,E2对HUVEC有抑制凋亡的作用,Hcy对HUVEC有促凋亡作用。选取最适浓度(1×10^-8μmol/L)E2和最适浓度(1×10^-4μmol/L)Hcy作用于细胞,实时定量PCR和Western blotting结果显示,E2组与E2+Hcy组MTHFR mRNA和蛋白的表达增加,而Hcy组无变化。结论E2促进HUVEC增殖,抑制HUVEC凋亡。Hcy抑制HUVEC增殖,促进HUVEC凋亡。E2和Hcy对HUVEC的作用可能与MTHFR的表达有关。
Objective To investigate the effects of estradiol(E2)and homocysteine(Hcy)on the proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs)and their underlying mechanism of action.Methods HUVECs were treated with different concentrations of E2 and Hcy,and proliferation of the treated cells was estimated by a cell counting kit-8(CCK-8)assay.The apoptosis rate of these cells was measured by a flow cytometric method employing annexin V-FITC/PI double staining.Optimal concentrations of E2 and Hcy were selected for treating HUVECs,and the control,E2,Hcy,and E2+Hcy groups were set.Expression levels of methylenetetrahydrofolate reductase(MTHFR)mRNA and the corresponding MTHFR protein in HUVECs were detected by real-time quantitative PCR and Western blotting,respectively.Results Results from the CCK-8 assay showed that E2 promoted the proliferation of HUVECs,whereas Hcy inhibited the proliferation.Insights obtained from flow cytometry demonstrated that the apoptosis of HUVECs was inhibited and facilitated by E2 and Hcy,respectively.The optimal concentrations of E2 and Hcy used for treating cells were 1×10^-8 and 1×10^-4μmol/L,respectively.Real-time quantitative PCR and Western blotting showed an increase in the expression of MTHFR mRNA and protein in E2 and E2+Hcy groups,while there was no change in expression levels in the Hcy group.Conclusion E2 promotes proliferation and inhibits apoptosis of HUVECs.Hcy exerts the opposite influence by inhibiting proliferation and promoting apoptosis of HUVECs.These opposing effects of E2 and Hcy on HUVECs might be related to the expression of MTHFR.
作者
陈海英
刘雨琪
CHEN Haiying;LIU Yuqi(Department of Obstetrics,The First Hospital,China Medical University,Shenyang 110001,China)
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2020年第5期429-432,436,共5页
Journal of China Medical University
基金
辽宁省自然科学基金(20180530005)。
关键词
雌二醇
同型半胱氨酸
人脐静脉内皮细胞
静脉血栓
estradiol
homocysteine
human umbilical vein endothelial cells
venous thrombosis
