中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (25): 4060-4064.doi: 10.12307/2024.184

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

环状RNA hsa-circ-0001360在同型半胱氨酸诱导人脐静脉内皮细胞凋亡中的作用

况园军1，2，于素美2，3，钟颖怡2，章旭红4，马胜超2，杨安宁2，郝银菊2，熊建团2，焦  运5，姜怡邓2   

  1. 宁夏医科大学，1检验学院，2国家卫生健康委代谢性心血管疾病研究重点实验室，3临床医学院，4基础医学院，宁夏回族自治区银川市   750004；5宁夏医科大学总医院，宁夏回族自治区银川市   750004
  • 收稿日期:2023-05-31 接受日期:2023-07-17 出版日期:2024-09-08 发布日期:2023-11-24
  • 通讯作者: 姜怡邓，博士，教授，宁夏医科大学国家卫生健康委代谢性心血管疾病研究重点实验室，宁夏回族自治区银川市 750004
  • 作者简介:况园军，男，1996年生，江西省宜春市人，汉族，宁夏医科大学在读硕士，主要从事心血管疾病形成的机制研究。
  • 基金资助:
    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

Effect of cyclic RNA hsa-circ-0001360 on homocysteine-induced apoptosis of human umbilical vein endothelial cells

Kuang Yuanjun1, 2, Yu Sumei2, 3, Zhong Yingyi2, Zhang Xuhong4, Ma Shengchao2, Yang Anning2, Hao Yinju2, Xiong Jiantuan2, Jiao Yun5, Jiang Yideng2   

  1. 1School of Inspection, 2Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, 3School of Clinical Medicine, 4Basical Medical College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 5General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2023-05-31 Accepted:2023-07-17 Online:2024-09-08 Published:2023-11-24
  • Contact: Jiang Yideng, MD, Professor, Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Kuang Yuanjun, Master candidate, School of Inspection, and Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    Fundamental Research Funds for Central Level Public Welfare Research Institutes of the Chinese Academy of Medical Sciences, No. 2019PT330002 (to JYD); National Natural Science Foundation of China, No. 82270492 (to MSC); National Natural Science Foundation of China, No. 82060090 (to HYJ); Key Research and Development Plan of Ningxia Hui Autonomous Region, No. 2022BFH02013 (to HYJ); Key Research and Development Plan of Ningxia Hui Autonomous Region, No. 2020BFH02003 (to YAN); Key Research and Development Plan of Ningxia Hui Autonomous Region, No. 2021BEG02033 (to XJT); Key Research and Development Plan of Ningxia Hui Autonomous Region, No. 2020BEG03005 (to JY)

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摘要：


文题释义：

人脐静脉内皮细胞：是来源于新生儿脐带静脉内壁的一种干细胞。因其容易获取、没有伦理学争议，且能够无限增殖，常作为内皮细胞功能和病理学的研究模型，应用于血管生成、愈合、血管建模、组织工程、炎症等研究中。
环状RNA：一种共价闭合的单链非编码RNA，其结构稳定性相较于其他RNA更强；环状RNA具有调控基因转录、调控靶蛋白和充当小分子RNA海绵等诸多功能。
同型半胱氨酸：是一种含硫氨基酸，是蛋氨酸代谢的中间产物，其本身不参与蛋白质的合成。高同型半胱氨酸血症是心脑血管病的新的、独立的危险因子。


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100 μmol/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。

https://orcid.org/0009-0000-9937-6056 (况园军) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 同型半胱氨酸, hsa-circ-0001360, 人脐静脉内皮细胞, 细胞凋亡, 环状RNA

Abstract: 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 100 μmol/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 hsa-circ-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. 

Key words: homocysteine, hsa-circ-0001360, human umbilical vein endothelial cell, apoptosis, cyclic RNA

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