2025年4月3日 周四
首页 > 按月查看>2016年第8月 >781-787
PDF 导出
坏死性凋亡与活性氧的相互作用介导高糖引起的H9c2心肌细胞损伤
作者:
作者单位:

(1.广州市番禺区中心医院心血管内科,;2.广州市番禺区心血管疾病研究所,广东省广州市 511400;3.中山大学附属第一医院黄埔院区心血管内科CCU,广东省广州市 510700;4.中山大学附属第一医院心血管内科,广东省广州市 510080)

作者简介:

梁伟杰,硕士,主治医师,研究方向为心血管介入和心血管疾病的保护机制,E-mail为279096515@qq.com。

基金项目:

国家自然科学基金资助项目(81270296)；广东省财政科技项目(2014SC107)


The Interaction Between Necroptosis and Reactive Oxygen Species Mediates High Glucose-induced Injury in H9c2 Cardiac Cells
Author:
Affiliation:

1.Department of Cardiology, Central Hospital of Panyu District;2.Cardiovascular Institute of Panyu District, Guangzhou, Guangdong 511400, China;3.Unit of Cardiac Care of Department of Cardiology, Huangpu Division of the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510700, China;4.Department of Cardiology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China)

  • 摘要
    • | |
  • 访问统计
    • |
  • 参考文献
    • |
  • 相似文献
    • | |
    摘要:

    目的 探讨坏死性凋亡(Nec)与活性氧(ROS)之间的相互作用能否介导高糖引起的H9c2 心肌细胞损伤。方法 应用蛋白质免疫印迹试验(Western blot)检测心肌细胞RIP3 蛋白的表达水平；细胞计数盒测定心肌细胞存活率；罗丹明 123(Rh123)染色荧光显微镜摄片测定线粒体膜电位(MMP)；2’, 7’-二氢二氯荧光素二乙酸酯(DCFH-DA)染色荧光显微镜摄片检测心肌细胞内ROS水平；Hoechst 33258核染色荧光显微镜摄片测定凋亡细胞的数量。结果 应用高糖(HG,35 mmol/L 葡萄糖)分别处理H9c2 细胞0～24 h,其中3、6、9、12和24 h均能明显地上调RIP3 蛋白的表达水平,24 h时RIP3 蛋白表达上调最为明显。应用100 μmol/L Nec的特异性阻断剂necrostatin-1(Nec-1)共处理心肌细胞可对抗高糖引起的RIP3 蛋白表达的上调作用。此外,100 μmol/L Nec-1能显著地抑制高糖引起的细胞毒性、线粒体损伤及氧化应激,使细胞存活率升高,MMP丢失及ROS生成减少。1 000 μmol/L ROS清除剂N-乙酰-L-半胱氨酸(NAC)预处理心肌细胞60 min,可明显地抑制高糖对心肌细胞RIP3 蛋白表达的上调作用；此外,高糖诱导的细胞凋亡和细胞毒性作用,经NAC预处理后均能得到显著的抑制。结论 Nec与ROS之间的相互作用介导高糖对心肌细胞的损伤。

    Abstract:

    Aim To investigate whether the interaction between necroptosis (Nec) and reactive oxygen species (ROS) mediates high glucose (HG)-induced injury in H9c2 cardiac cells. Methods The expression level of RIP3 protein (an important index that reflects Nec) was determined by Western blot assay. The cell viability was measured by cell counting kit-8 (CCK-8) assay. Mitochondrial membrane potential (MMP) was examined by Rhodamine 123(Rh 123)staining followed by photofluorography. The intracellular levels of ROS were detected by 2’, 7’-dichlorfluorescein-diacetate (DCFH-DA) staining followed by photofluorography. Apoptotic cells were evaluated by the nuclear morphology observed with Hoechst 33258 staining followed by photofluorography. Results After the H9c2 cells were treated with 35 mmol/L glucose (HG) for 0～24 h, respectively, the expression levels of RIP3 protein were significantly increased at 3 h, 6 h, 9 h, 12 h and 24 h, reaching the maximum level at 24 h. Cotreatment of the cells with necrostatin-1 (Nec-1, a specific inhibitor of Nec) considerably blocked the up-regulation of RIP3 expression level induced by HG. Moreover, cotreatment with Nec-1 obviously inhibited HG-induced injuries (including cytotoxicity, mitochondrial damage and oxidative stress), leading to an increase in the cell viability, decreases in a loss of MMP and ROS generation. On the other hand, pretreatment of the cells with 1000 μm N-acetyl-L-cysteine (NAC, a scavenger of ROS) for 60 min before HG exposure obviously reduced the HG-induced increase in the expression of RIP3. In addition, pretreatment of the cells with NAC dramatically alleviated the HG-induced apoptosis and cytotoxicity. Conclusion The interaction between Nec and ROS mediates HG-induced injury in H9c2 cardiac cells.

    参考文献
    [1] Cai L, Kang YJ.Cell death and diabetic cardiomyopathy.Cardiovasc Toxicol, 3,3(3): 219-228.
    [2] Nunes T, Bernardazzi C, de Souza HS.Cell death and inflammatory bowel diseases: apoptosis, necrosis, and autophagy in the intestinal epithelium.Biomed Res Int, 4,4: 218-493.
    [3] Fenton K.The effect of cell death in the initiation of lupus nephritis.lin Exp Immunol, 5,9(1): 11-16.
    [4] Golstein P, Kroemer G.Cell death by necrosis: towards a molecular definition.Trends Biochem Sci, 7,2(1): 37-43.
    [5] Luedde M, Lutz M, Carter N, et al.RIP3, a kinase promoting necroptotic cell death, mediates adverse remodelling after myocardial infarction.Cardiovasc Res.4,3(2): 206-216.
    [6] Degterev A, Huang Z, Boyce M, et al.Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury.Nat Chem Biol.5,1(2): 112-119.
    [7] Cho YS, Challa S, Moquin D, et al.Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation.Cell, 9,7(6): 1 112-123.
    [8] Degterev A, Zhou W, Maki JL, et al.Assays for necroptosis and activity of RIP kinases.Methods Enzymol, 4,5: 1-33.
    [9] Liu YS, Huang ZW, Wang L, et al.Sitagliptin alleviated myocardial remodeling of the left ventricle and improved cardiac diastolic dysfunction in diabetic rats.J Pharmacol Sci, 5,7(3): 260-274.
    [10] Schenk B, Fulda S.Reactive oxygen species regulate Smac mimetic/TNFα-induced necroptotic signaling and cell death.Oncogene, 5,4(47): 5 796-806.
    [11] Xu W, Wu W, Chen J, et al.Exogenous hydrogen sulfide protects H9c2 cardiac cells against high glucose-induced injury by inhibiting the activities of the p38 MAPK and ERK1/2 pathways.Int J Mol Med, 3,2(4): 917-925.
    [12] Liu C, Wang L, He K.Diabetic cardiomyopathy and autophagy.Zhonghua Bing Li Xue Za Zhi, 5,4(2): 146-149.
    [13] Ouyang C, You J, Xie Z.The interplay between autophagy and apoptosis in the diabetic heart.J Mol Cell Cardiol, 4,1: 71-80.
    [14] Nunes T, Bernardazzi C, de Souza HS.Cell death and inflammatory bowel diseases: apoptosis, necrosis, and autophagy in the intestinal epithelium.Biomed Res Int, 4,4: 218 493.
    [15] Zhuang XD, Hu X, Long M, et al.Exogenous hydrogen sulfide alleviates high glucose-induced cardiotoxicity via inhibition of leptin signaling in H9c2 cells.Mol Cell Biochem.4,1(1-2): 147-155.
    [16] 梁伟杰,陈景福,张稳柱,等.ATP敏感性钾通道在硫化氢抑制高糖引起的心肌细胞损伤中的作用.中国病理生理杂志, 5,1(5): 785-790.
    [17] Xu W, Chen J, Lin J, et al.Exogenous H2S protects H9c2 cardiac cells against high glucose-induced injury and inflammation by inhibiting the activation of the NF-κB and IL-1β pathways.Int J Mol Med, 5,5(1): 177-186.
    [18] 梁伟杰,陈景福,宋明才,等.活性氧与ATP敏感性钾通道的相互作用参与高糖对H9c2心肌细胞的损伤.中国动脉硬化杂志, 5,3(11): 1 081-088.
    [19] 佘美华,蒋文艳,张瑶,等.褪黑素通过抑制ROS改善胰岛素抵抗HepG2细胞糖代谢.中南医学科学杂志, 4,2(5): 433-435.
    相似文献
    引证文献
引用本文

梁伟杰,何洁仪,陈景福,陈君,余盛龙,宋明才,郑东诞,廖新学,张稳柱.坏死性凋亡与活性氧的相互作用介导高糖引起的H9c2心肌细胞损伤[J].中国动脉硬化杂志,2016,24(8):781~787.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2015-12-28
  • 最后修改日期:2016-02-18
  • 在线发布日期: 2016-07-25

您是本站第 4759234 访问者

邮编：421001 传真：0734-8160523

电话：0734-8160765 E-mail：dmzzbjb@163.net

中国动脉硬化杂志 ® 2025 网站版权所有