麦冬多糖对转化生长因子-β1诱导心肌细胞凋亡的影响
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摘要
目的探讨麦冬多糖(MDG-1)对转化生长因子(TGF)-β1诱导的心肌细胞活力、凋亡的影响及机制。方法大鼠H9c2心肌细胞分为对照组(不经特殊处理)、TGF-β1组(20 ng/ml TGF-β1处理细胞)和MDG-1组(100 mg/L MDG-1及20 ng/ml TGF-β1处理细胞),噻唑蓝(MTT)及流式细胞术分别检测细胞活力及凋亡率;2′,7′-二氯荧光黄双乙酸盐(DCFH-DA)探针检测活性氧(ROS)水平;Western印迹检测增殖细胞抗原(PCNA)、B淋巴细胞瘤(Bcl)-2、p53、核因子(NF)-κB p65、KB抑制蛋白激酶(IKK)α和β-catenin的蛋白表达。结果与对照组比较,TGF-β1组细胞活力显著降低,PCNA和Bcl-2蛋白表达显著降低,细胞凋亡率显著升高,ROS水平显著升高,p53、NF-κB p65、IKKα和β-catenin蛋白表达显著升高(P<0.05)。与TGF-β1组比较,MDG-1组细胞活力显著升高,PCNA和Bcl-2蛋白表达显著升高,细胞凋亡率显著降低,ROS水平显著降低,p53、NF-κB p65、IKKα和β-catenin蛋白表达显著降低(P<0.05)。结论 MDG-1可增强心肌细胞活力,抑制心肌细胞凋亡,其机制可能与降低细胞内ROS水平及抑制Wnt/β-catenin信号和NF-κB信号有关。
Objective To investigate the effect and mechanism of ophiopogon japonicus polysaccharide(MDG-1) on TGF-β1 induced cardiomyocyte viability and apoptosis. Methods H9 c2 rat cardiomyocytes were divided into three treatment groups, namely control group(without special treatment), TGF-β1 group(20 ng/ml TGF-β1 treated cells) and MDG-1 group(100 mg/L MDG-1 and 20 ng/ml TGF-β1 treated cells), cell viability and apoptosis rate were detected by MTT and flow cytometry respectively; DCFH-DA probe was used to detect ROS level; Western blot was used to detect the expressions of PCNA, Bcl-2, p53, NF-κB p65, IKKα and β-catenin proteins. Results Compared with that of control group, the cell viability of TGF-β1 group was significantly decreased, the expressions of PCNA and Bcl-2 proteins were significantly decreased, the apoptosis rate,the ROS level, and the expressions of p53, NF-κB p65, IKKα and β-catenin proteins were significantly increased(P<0.05). Compared with those of TGF-β1 group, the cell viability of the MDG-1 group was significantly increased, the expressions of PCNA and Bcl-2 proteins were significantly increased, the apoptosis rate was significantly decreased, and the ROS level was significantly decreased, the expressions of p53, NF-κB p65, IKKα and β-catenin proteins were significantly decreased(P<0.05).Conclusions MDG-1 could enhance the activity of cardiomyocytes and inhibit the apoptosis of cardiomyocytes. The mechanism might be related to the decrease of intracellular ROS level and the inhibition of Wnt/β-catenin signal and NF-κB signal.
Objective To investigate the effect and mechanism of ophiopogon japonicus polysaccharide(MDG-1) on TGF-β1 induced cardiomyocyte viability and apoptosis. Methods H9 c2 rat cardiomyocytes were divided into three treatment groups, namely control group(without special treatment), TGF-β1 group(20 ng/ml TGF-β1 treated cells) and MDG-1 group(100 mg/L MDG-1 and 20 ng/ml TGF-β1 treated cells), cell viability and apoptosis rate were detected by MTT and flow cytometry respectively; DCFH-DA probe was used to detect ROS level; Western blot was used to detect the expressions of PCNA, Bcl-2, p53, NF-κB p65, IKKα and β-catenin proteins. Results Compared with that of control group, the cell viability of TGF-β1 group was significantly decreased, the expressions of PCNA and Bcl-2 proteins were significantly decreased, the apoptosis rate,the ROS level, and the expressions of p53, NF-κB p65, IKKα and β-catenin proteins were significantly increased(P<0.05). Compared with those of TGF-β1 group, the cell viability of the MDG-1 group was significantly increased, the expressions of PCNA and Bcl-2 proteins were significantly increased, the apoptosis rate was significantly decreased, and the ROS level was significantly decreased, the expressions of p53, NF-κB p65, IKKα and β-catenin proteins were significantly decreased(P<0.05).Conclusions MDG-1 could enhance the activity of cardiomyocytes and inhibit the apoptosis of cardiomyocytes. The mechanism might be related to the decrease of intracellular ROS level and the inhibition of Wnt/β-catenin signal and NF-κB signal.
引文
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20 Venkatesan B,Prabhu SD,Venkatachalam K,et al.WNT1-inducible signaling pathway protein-1 activates diverse cell survival pathways and blocks doxorubicin-induced cardiomyocyte death[J].Cell Signalling,2010;22(5):809-20.
21 Zhao T,Tang X,Umeshappa CS,et al.Simulated microgravity promotes cell apoptosis through suppressing Uev1A/TICAM/TRAF/NF-κB-Regulated anti-apoptosis and p53/PCNA- and ATM/ATR-Chk1/2-Controlled DNA-damage response pathways[J].J Cell Biochem,2016;117(9):2138-48.
2 Lin Z,Song D,Wei H,et al.TGF-β1-induced miR-202 mediates drug resistance by inhibiting apoptosis in human osteosarcoma[J].J Cancer Res Clin Oncol,2016;142(1):239-46.
3 Wei Q,Xin C,Liu P.Inhibition of TGF-β1 by eNOS gene transfer provides cardiac protection after myocardial infarction[J].J Biomed Res,2010;24(2):145-52.
4 Wang Y,Zong L,Wang X.TGF-β improves myocardial function and prevents apoptosis induced by anoxia-reoxygenation,through the reduction of endoplasmic reticulum stress[J].Canadian J Physiol Pharmacol,2015;94(1):9-17.
5 沈宇腾,张羽飞,顾冬雪,等.FGF-1对TGF-β1诱导的H9c2心肌细胞凋亡的影响及机制研究[J].牡丹江医学院学报,2017;38(1):4-7.
6 许闪,王靓,黄金玲,等.苓桂术甘汤含药血清对TGF-β1诱导的大鼠心肌细胞株H9c2凋亡的影响[J].中药药理与临床,2016;32(3):4-8.
7 沙建平,马红英,陈晓文,等.麦冬对糖尿病大鼠胰岛β细胞的保护作用[J].成都中医药大学学报,2014;37(3):23-4.
8 王硕.麦冬多糖MDG-1抗心肌缺血作用机制研究[D].上海:上海中医药大学,2009.
9 Ke ZP,Peng X,Yan S,et al.MicroRNA-93 inhibits ischemia-reperfusion induced cardiomyocyte apoptosis by targeting PTEN[J].Oncotarget,2016;7(20):28796-805.
10 Jia Z,Wang J,Shi Q,et al.SOX6andPDCD4enhance cardiomyocyte apoptosis through LPS-induced miR-499 inhibition[J].Apoptosis,2016;21(2):174-83.
11 苏萍,吕书峰,范雪梅,等.参麦注射液及其有效组分对H202诱导心肌细胞损伤的保护作用[J].中成药,2011;33(12):2150-3.
12 Song W,Wang X.The role of TGFβ1 and LRG1 in cardiac remodelling and heart failure[J].Biophys Rev,2015;7(1):91-104.
13 Liu ZW,Zhu HT,Chen KL,et al.Selenium attenuates high glucose-induced ROS/TLR-4 involved apoptosis of rat cardiomyocyte[J].Biolog Trace Element Res,2013;156(1-3):262-70.
14 Lu H,Tian A,Wu J,et al.Danshensu inhibits β-adrenergic receptors-mediated cardiac fibrosis by ROS/p38 MAPK axis[J].Biolog Pharmaceut Bull,2014;37(6):961-7.
15 Gao D,Yang J,Wu Y,et al.Targeting dynamin 2 as a novel pathway to inhibit cardiomyocyte apoptosis following oxidative stress[J].Cell Physiol Biochem,2016;39(6):2121-34.
16 Zhang Z,Zhou C,Chang Y,et al.Long non-coding RNA CASC11 interacts with hnRNP-K and activates the WNT/β-catenin pathway to promote growth and metastasis in colorectal cancer[J].Cancer Lett,2016;376(1):62-73.
17 Ye B,Hou N,Xiao L,et al.APC controls asymmetric Wnt/β-catenin signaling and cardiomyocyte proliferation gradient in the heart[J].J Mol Cell Cardiol,2015;89(Pt B):287-96.
18 周浩,王广庆,罗鹏飞,等.核因子κB活化在脓毒症所致心肌损伤中的作用[J].中华烧伤杂志,2017;33(12):782-4.
19 黄小芳,张韬,华玥,等.Wnt/β-连环蛋白在高糖诱导心肌成纤维细胞增殖中的作用[J].贵阳医学院学报,2014;39(1):30-4.
20 Venkatesan B,Prabhu SD,Venkatachalam K,et al.WNT1-inducible signaling pathway protein-1 activates diverse cell survival pathways and blocks doxorubicin-induced cardiomyocyte death[J].Cell Signalling,2010;22(5):809-20.
21 Zhao T,Tang X,Umeshappa CS,et al.Simulated microgravity promotes cell apoptosis through suppressing Uev1A/TICAM/TRAF/NF-κB-Regulated anti-apoptosis and p53/PCNA- and ATM/ATR-Chk1/2-Controlled DNA-damage response pathways[J].J Cell Biochem,2016;117(9):2138-48.