摘要
背景:同型半胱氨酸水平上升会诱导人脐静脉内皮细胞发生凋亡,但机制尚不清楚。目的:探讨hsa-circ-0001360在同型半胱氨酸诱导人脐静脉内皮细胞发生凋亡中的作用。方法:体外培养人脐静脉内皮细胞,将其分为对照组、同型半胱氨酸组、干扰对照组、干扰对照+同型半胱氨酸组、干扰hsa-circ-0001360组、干扰hsa-circ-0001360+同型半胱氨酸组、过表达对照组、过表达对照+同型半胱氨酸组、过表达hsa-circ-0001360组和过表达hsa-circ-0001360+同型半胱氨酸组,同型半胱氨酸的干预浓度均为100μmol/L。干预细胞72 h后,应用Western blot检测凋亡相关蛋白Bax、Bcl-2和Caspase-3的表达;流式细胞仪检测细胞的凋亡率;实时荧光定量PCR(qRT-PCR)检测hsa-circ-0001360的表达水平。结果与结论:①与对照组相比,同型半胱氨酸组人脐静脉内皮细胞中Caspase-3和Bax的表达明显升高(P<0.01),Bcl-2的表达明显降低(P<0.01),细胞凋亡率明显增高(P<0.01);②与对照组相比,同型半胱氨酸组人脐静脉内皮细胞中hsa-circ-0001360的表达明显升高(P<0.01);③hsa-circ-0001360在人脐静脉内皮细胞的细胞质中表达显著高于细胞核(P<0.01);④与干扰对照组或干扰对照+同型半胱氨酸组相比,干扰hsa-circ-0001360组和干扰hsa-circ-0001360+同型半胱氨酸组人脐静脉内皮细胞中Caspase-3、Bax的表达明显降低(P<0.01),Bcl-2的表达明显升高(P<0.01),细胞凋亡率明显降低(P<0.01);⑤与过表达对照组或过表达对照+同型半胱氨酸组相比,过表达hsa-circ-0001360组和过表达hsa-circ-0001360+同型半胱氨酸组人脐静脉内皮细胞中Caspase-3、Bax的表达明显升高(P<0.01),Bcl-2的表达明显降低(P<0.01),细胞凋亡率明显升高(P<0.01);⑥以上结果表明,hsa-circ-0001360可以促进同型半胱氨酸诱导人脐静脉内皮细胞发生凋亡。
BACKGROUND:Increased homocysteine level induces apoptosis of human umbilical vein endothelial cells,but the mechanism remains unclear.OBJECTIVE:To investigate the role of hsa-circ-0001360 in human umbilical vein endothelial cell apoptosis induced by homocysteine.METHODS:In vitro cultured human umbilical vein endothelial cells were divided into control group,homocysteine group,interference control group,interference control+homocysteine group,hsa-circ-0001360 interference group,hsa-circ-0001360+homocysteine interference group,overexpression control group,overexpression control+homocysteine group,hsa-circ-0001360 overexpression group and hsa-circ-0001360+homocysteine overexpression group.All groups were treated with 100μmol/L homocysteine.After 72 hours of intervention,the expressions of apoptosis-related proteins Bax,Bcl-2,and Caspase-3 were detected by western blot assay.The apoptotic rate was detected by flow cytometry.Quantitative real-time PCR was used to detect the expression of hsacirc-0001360.RESULTS AND CONCLUSION:(1)Compared with the control group,the expression of Caspase-3 and Bax was significantly increased(P<0.01),and the expression of Bcl-2 was significantly decreased(P<0.01),and the apoptotic rate was significantly increased(P<0.01)in the homocysteine group.(2)Compared with control group,the expression of hsa-circ-0001360 was significantly increased in the homocysteine group(P<0.01).(3)The expression of hsa-circ-0001360 was significantly higher in the cytoplasm than that in the nucleus(P<0.01).(4)Compared with the interference control C group and interference control+homocysteine group,the expressions of Caspase-3 and Bax were significantly decreased(P<0.01),while the expression of Bcl-2 was significantly increased(P<0.01);the apoptotic rate was significantly decreased(P<0.01)in sh-hsa-circ-0001360 interference group and sh-hsa-circ-0001360+homocysteine interference group.(5)Compared with overexpression control group and overexpression control+homocysteine group,the expressions of Caspase-3 and Bax were significantly increased(P<0.01),while the expression of Bcl-2 was significantly decreased(P<0.01);the apoptotic rate was significantly increased(P<0.01)in the hsa-circ-0001360 overexpression group and the hsa-circ-0001360+homocysteine overexpression group.(6)In conclusion,hsa-circ-0001360 can promote the apoptosis of human umbilical vein endothelial cells induced by homocysteine.
作者
况园军
于素美
钟颖怡
章旭红
马胜超
杨安宁
郝银菊
熊建团
焦运
姜怡邓
Kuang Yuanjun;Yu Sumei;Zhong Yingyi;Zhang Xuhong;Ma Shengchao;Yang Anning;Hao Yinju;Xiong Jiantuan;Jiao Yun;Jiang Yideng(School of Inspection,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Basical Medical College,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第25期4060-4064,共5页
Chinese Journal of Tissue Engineering Research
基金
中国医学科学院中央级公益性科研院所基本科研业务费项目(2019PT330002),项目负责人:姜怡邓
国家自然科学基金(82270492),项目负责人:马胜超
国家自然科学基金(82060090),项目负责人:郝银菊
宁夏回族自治区重点研发计划(2022BFH02013),项目负责人:郝银菊
宁夏回族自治区重点研发计划(2020BFH02003),项目负责人:杨安宁
宁夏回族自治区重点研发计划(2021BEG02033),项目负责人:熊建团
宁夏回族自治区重点研发计划(2020BEG03005),项目负责人:焦运。
