中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (23): 3621-3627.doi: 10.12307/2023.575

• 血管组织构建 vascular tissue construction • 上一篇    下一篇

松弛素抑制氧化应激反应保护小鼠心肌微血管内皮细胞的缺氧/复氧损伤

魏  琴1，2，陈冰心3，赵  翎4，阿曼古丽·如则4，赵帮豪4，姜  涛2，4，张  春2，4，李志强2，4，段明军4，高晓明2   

  1. 新疆医科大学，1中心实验室，4动物实验中心，新疆维吾尔自治区乌鲁木齐市  830054；2新疆医学动物模型研究重点实验室，新疆维吾尔自治区乌鲁木齐市  830054；3新疆医科大学第一附属医院心功能科，新疆维吾尔自治区乌鲁木齐市  830054
  • 收稿日期:2022-06-29 接受日期:2022-09-24 出版日期:2023-08-18 发布日期:2023-01-14
  • 通讯作者: 段明军，硕士，助理研究员，新疆医科大学动物实验中心，新疆维吾尔自治区乌鲁木齐市 830054 高晓明，博士，教授，研究员，新疆医学动物模型研究重点实验室，新疆维吾尔自治区乌鲁木齐市 830054
  • 作者简介:魏琴，女，1980年生，硕士，实验师，主要从事心血管疾病基础研究。 陈冰心，女，1984年生，硕士，主治医师，主要从事心血管疾病临床研究。
  • 基金资助:
    新疆维吾尔自治区自然科学基金(2018D01C197)，项目负责人：段明军；新疆维吾尔自治区重点实验室开放课题
    (2021D04020)，项目负责人：高晓明；省部共建中亚高发病成因与防治国家重点实验室心血管专项(SKL-HIDCA-2021-XXG1)，
    项目负责人：阿曼古丽·如则

Serelaxin protects against hypoxia/reoxygenation-induced mouse cardiac microvascular endothelial cell injury by inhibiting oxidative stress responses

Wei Qin1, 2, Chen Bingxin3, Zhao Ling4, Amanguli·Ruze4, Zhao Banghao4, Jiang Tao2, 4, Zhang Chun2, 4, Li Zhiqiang2, 4, Duan Mingjun4, Gao Xiaoming2   

  1. 1Center Laboratory, 4Animal Laboratory Center, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Xinjiang Key Laboratory of Medical Animal Model Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 3Department of Cardiac Function, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2022-06-29 Accepted:2022-09-24 Online:2023-08-18 Published:2023-01-14
  • Contact: Duan Mingjun, Master, Assistant researcher, Animal Laboratory Center, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China Gao Xiaoming, MD, Professor, Researcher, Xinjiang Key Laboratory of Medical Animal Model Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Wei Qin, Master, Experimentalist, Center Laboratory, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Key Laboratory of Medical Animal Model Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China Chen Bingxin, Master, Attending physician, Department of Cardiac Function, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 2018D01C197 (to DMJ); the Open Project of the Key Laboratory of Xinjiang Uygur Autonomous Region, No 2021D04020 (to GXM); the Special Project of Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, No. SKL-HIDCA-2021-XXG1 (to ARZ)

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摘要：

文题释义：

松弛素：作为一种新型的肽类激素，具有抗炎、细胞外基质重构效应、维持内皮功能、血管重塑及显著的抗纤维化能力和其他心脏保护作用，同时可减少终末性心力衰竭的风险。
心肌缺血再灌注损伤：心肌缺血后再通使其重新获得血供时，由于再灌注时的氧化应激、炎症反应、钙超载和pH值异常等多种因素，可能引起心脏超微结构、功能、代谢及电生理方面进一步损伤，是在缺血损伤基础上再次引起的损伤。

背景：松弛素是人松弛素2的重组形式，作为机体内源性抗纤维化活性物质，松弛素治疗可能会缓解急性心力衰竭患者的症状并改善其预后，但尚未有研究松弛素在氧化应激方面对缺氧/复氧损伤小鼠心肌微血管内皮细胞(H5V)的作用。
目的：探讨松弛素在小鼠H5V细胞缺氧/复氧损伤模型中的作用。
方法：建立小鼠H5V细胞缺氧/复氧损伤模型，分别为缺氧3，6，12 h，复氧3 h，通过检测细胞活力和乳酸脱氢酶活性确定细胞缺氧/复氧的最佳时间。选用不同质量浓度的松弛素(0，60，100，140，180 ng/mL)作用于H5V细胞缺氧/复氧损伤模型，通过检测细胞活力和乳酸脱氢酶活性确定松弛素干预的最佳质量浓度。在此基础上，将H5V细胞分为3组：对照组、缺氧/复氧模型组、缺氧/复氧+180 ng/mL松弛素组，检测丙二醛、超氧化物歧化酶和活性氧水平。
结果与结论：①与对照组比较，缺氧6 h/复氧3 h、缺氧12 h/复氧3 h的细胞活力均显著降低(P < 0.05)，细胞上清中乳酸脱氢酶活性均显著增加(P < 0.05)，选取缺氧6 h/复氧3 h进行后续实验；②与缺氧/复氧模型组比较，缺氧/复氧+松弛素180 ng/mL组细胞活力显著增加(P < 0.05)，细胞上清中乳酸脱氢酶活性显著降低(P < 0.05)，选取松弛素180 ng/mL进行后续干预实验；③与缺氧/复氧模型组比较，缺氧/复氧+180 ng/mL松弛素组活性氧、丙二醛水平显著降低(P < 0.05)，超氧化物歧化酶水平显著增加(P < 0.05)；④结果表明，H5V细胞经缺氧6 h/复氧3 h可建立H5V细胞损伤模型，经180 ng/mL松弛素干预后提高了细胞活性和细胞抗氧化应激能力。

https://orcid.org/0000-0003-0344-8305(魏琴)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 松弛素, 小鼠心肌微血管内皮细胞, 缺血再灌注损伤, 丙二醛, 超氧化物歧化酶, 活性氧, 氧化应激反应

Abstract: BACKGROUND: Serelaxin is a recombinant form of human relaxin 2. As endogenous anti-fibrosis active substances, serelaxin may relieve the symptoms and improve the prognosis of patients with acute heart failure. However, there is no report on the effect of serelaxin on hypoxia/reoxygenation injury of myocardial microvascular endothelial cells (H5V) in mice through regulating oxidative stress.
OBJECTIVE: To investigate the role on serelaxin in a model of hypoxia/reoxygenation-induced mouse cardiac microvascular endothelial cells (H5V)injury.
METHODS: To establish a mouse H5V hypoxia/reoxygenation injury model, cells were incubated under hypoxia for 3, 6, and 12 hours followed by 3 hours of reoxygenation. The optimal time of cellular hypoxia/reoxygenation was determined by cell viability and lactate dehydrogenase activity assay. Different concentrations of serelaxin (0, 60, 100, 140, 180 ng/mL) were selected to act in the H5V cellular hypoxia/reoxygenation injury model, and the optimal drug concentration for drug treatment of cellular hypoxia-reoxygenation injury was determined by the cell viability and lactate dehydrogenase activity assays. H5V cells were then divided into three groups: control, model (hypoxia/reoxygenation), and serelaxin (hypoxia/reoxygenation+180 ng/mL serelaxin) groups. Malondialdehyde, superoxide dismutase and reactive oxygen species levels were detected in each group.
RESULTS AND CONCLUSION: Compared with the control group, cell proliferation was significantly reduced after hypoxia 6 hours/reoxygenation 3 hours and hypoxia 12 hours/reoxygenation 3 hours (P < 0.05), while the lactate dehydrogenase activity in cell supernatant was significantly increased (P < 0.05). Therefore, cells cultured under hypoxia 6 hours/reoxygenation 3 hours were selected for subsequent experiments. Cell viability was significantly increased in the serelaxin group compared with the model group (P < 0.05), while the lactate dehydrogenase activity in cell supernatant was significantly decreased (P < 0.05). Therefore, 180 ng/mL serelaxin was selected for subsequent experiments. Compared with the model group, the serelaxin group showed a significant decrease in reactive oxygen species and malondialdehyde levels but a significant increase in the level of superoxide dismutase (all P < 0.05). To conclude, H5V cells subjected to hypoxia 6 hours/reoxygenation 3 hours could establish a model of H5V vascular endothelial cell injury, and 180ng/mL serelaxin could improve the cell viability and anti-oxidative stress ability. 

Key words: serelaxin, mouse cardiac microvascular endothelial cell, Ischemia-reperfusion injury, malondialdehyde, superoxide dismutase, reactive oxygen species, oxidative stress response

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