榅桲总黄酮对心肌梗死大鼠心肌损伤的保护作用及对JNK和NF-κB通路的影响
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摘要
目的探究榅桲总黄酮(COMF)对心肌梗死大鼠心肌损伤的保护作用及机制。方法构建急性心肌梗死大鼠模型,随机分为5组,每组20只,sham组和模型组灌胃给蒸馏水10μL/g;阿托伐他汀钙片(ACT)组灌胃给予ACT 8mg/kg;榅桲总黄酮低剂量组(L-COMF)和榅桲总黄酮高剂量组(H-COMF)组分别灌胃给予榅桲总黄酮80μg/g和160μg/g,连续给药3周。氯化三苯基四氮唑红(TTC)染色法检测心肌梗死面积;血流动力学检测心功能;苏木精-伊红(HE)染色检测组织学变化;TUNEL法检测心肌细胞凋亡;酶链免疫吸附试验(ELISA)检测肿瘤坏死因子(TNF)-α、单核细胞趋化蛋白-1(MCP-1)水平;Western blot分析核因子(NF)-κB和丝裂原活化蛋白激酶(MAPKs)活性。结果与sham组相比,模型组大鼠心肌梗死面积、TUNEL阳性细胞数、TNF-α和MCP-1水平、NF-κB、p-ERK1/2、p-C-Jun N-末端激酶(JNK)、p-p38 MAPK、Bax蛋白表达显著升高,Bcl-2蛋白表达显著降低(P<0.05)。与模型组相比,ACT组、L-COMF组、H-COMF组大鼠心肌梗死面积、TUNEL阳性细胞数、TNF-α和MCP-1水平、NF-κB、p-JNK、Bax蛋白表达显著降低,Bcl-2蛋白表达显著升高(P<0.05)。与sham组相比,模型组大鼠平均动脉压(MAP)、左心室收缩压(LVSP)显著降低,左心室舒张末压(LVEDP)显著升高,同时左心室内压上长最大速率(LV+dp/dtmax)显著降低,左心室内压下降最大速率(LV-dp/dtmax)显著升高(P<0.05)。与模型组相比,ACT组、L-COMF组、HCOMF组大鼠MAP、LVSP显著升高,LVEDP显著降低,同时LV+dp/dtmax显著升高,LV-dp/dtmax显著降低(P<0.05)。结论榅桲总黄酮能够对心肌梗死大鼠心肌损伤发挥保护作用,这可能是通过抑制JNK和NF-κB信号通路,从而下调心肌梗死引起的炎症损伤和细胞凋亡来实现的。
Objective To investigate the protective effect of total flavonoids of Cydonia Oblonga Mill(COMF)on myocardial injury in myocardial infarction in rats and its possible mechanism.Methods Rat model of acute myocardial infarction was established and randomly divided into 5 groups,20 rats in each group.The sham group and model group were given the intragastric administration of distilled water 10μL/g.The atorvastatin calcium tablets(ACT)group was given orally atorvastatin 8 mg/kg.The low-dose COMF(L-COMF)group and high-dose COMF(H-COMF)group were given COMF as 80μg/g and 160μg/g respectively,continuous intragastric administration for 3 weeks.The area of myocardial infarction was determined by TTC staining.And cardiac function was determined by hemodynamic detection.Histological changes were detected by hematoxylin-eosin(HE)staining.TUNEL assay was used to detect the apoptosis of myocardial cells.Enzyme chain immunosorbent assay(ELISA)was used to detect the tumor necrosis factor-α(TNF-α)and monocyte chemokine protein-1(MCP-1)levels.Western blot was performed to analyze the activities of nuclear factor-κB(NF-κB)and mitogen-activated protein kinase(MAPKs).Results Compared with the sham group,area of myocardial infarction,TUNEL positive cells,TNF-αand MCP-1,NF-κB,p-ERK1/2,p-JNK,p-p38 MAPKs,Bax expression in the ACT,L-COMF and H-COMF group significantly increased,Bcl-2 protein expression significantly decreased(P<0.05).Compared with the model group,area of myocardial infarction,TUNEL positive cells,TNF-αand MCP-1,NF-κB,p-ERK1/2,p-JNK,Bax expression in the ACT,L-COMF and H-COMF group significantly decreased,Bcl-2 protein expression significantly increased(P<0.05).Compared with the sham group,mean arterial pressure(MAP),left ventricular systolic pressure(LVSP)and LV+dp/dtmax significantly decreased,while LV-dp/dtmax were significantly increased(P<0.05).Compared with the model group,MAP,LVSP and LV+dp/dtmax significantly increased,while LVEDP and LV-dp/dtmax significantly decreased(P<0.05).Conclusion COMF plays a protective role in the myocardial injury of myocardial infarction rats,which may be through inhibition of JNK and NF-κB signaling pathways and down-regulating the inflammatory injury and cell apoptosis induced by myocardial infarction.
Objective To investigate the protective effect of total flavonoids of Cydonia Oblonga Mill(COMF)on myocardial injury in myocardial infarction in rats and its possible mechanism.Methods Rat model of acute myocardial infarction was established and randomly divided into 5 groups,20 rats in each group.The sham group and model group were given the intragastric administration of distilled water 10μL/g.The atorvastatin calcium tablets(ACT)group was given orally atorvastatin 8 mg/kg.The low-dose COMF(L-COMF)group and high-dose COMF(H-COMF)group were given COMF as 80μg/g and 160μg/g respectively,continuous intragastric administration for 3 weeks.The area of myocardial infarction was determined by TTC staining.And cardiac function was determined by hemodynamic detection.Histological changes were detected by hematoxylin-eosin(HE)staining.TUNEL assay was used to detect the apoptosis of myocardial cells.Enzyme chain immunosorbent assay(ELISA)was used to detect the tumor necrosis factor-α(TNF-α)and monocyte chemokine protein-1(MCP-1)levels.Western blot was performed to analyze the activities of nuclear factor-κB(NF-κB)and mitogen-activated protein kinase(MAPKs).Results Compared with the sham group,area of myocardial infarction,TUNEL positive cells,TNF-αand MCP-1,NF-κB,p-ERK1/2,p-JNK,p-p38 MAPKs,Bax expression in the ACT,L-COMF and H-COMF group significantly increased,Bcl-2 protein expression significantly decreased(P<0.05).Compared with the model group,area of myocardial infarction,TUNEL positive cells,TNF-αand MCP-1,NF-κB,p-ERK1/2,p-JNK,Bax expression in the ACT,L-COMF and H-COMF group significantly decreased,Bcl-2 protein expression significantly increased(P<0.05).Compared with the sham group,mean arterial pressure(MAP),left ventricular systolic pressure(LVSP)and LV+dp/dtmax significantly decreased,while LV-dp/dtmax were significantly increased(P<0.05).Compared with the model group,MAP,LVSP and LV+dp/dtmax significantly increased,while LVEDP and LV-dp/dtmax significantly decreased(P<0.05).Conclusion COMF plays a protective role in the myocardial injury of myocardial infarction rats,which may be through inhibition of JNK and NF-κB signaling pathways and down-regulating the inflammatory injury and cell apoptosis induced by myocardial infarction.
引文
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[17]张云,旦菊花,孙鑫,等.MAPKs信号通路在动脉粥样硬化发生发展中的调控作用[J].现代生物医学进展,2017,17(5):968-970.
[18]KIM E K,CHOI E J.Compromised MAPK signaling in human diseases:an update[J].Arch Toxicol,2015,89(6):867-82.
[19]呼雅丽.黄癸固体分散体通过抑制JNK保护心肌缺血再灌注损伤的作用机制研究[D].吉林大学,2015.
[20]张腾,张艳军,庄朋伟,等.瓜蒌对心梗后心衰大鼠心功能及心肌细胞凋亡的影响[J].中国药理学通报,2016,32(8):1183-1184.
[21]GUO C,YANG M,JING L,et al.Amorphous silica nanoparticles trigger vascular endothelial cell injury through apoptosis and autophagy via reactive oxygen species-mediated MAPK/Bcl-2and PI3K/Akt/mTOR signaling[J].Int J Nanomedicine,2016,11:5257-5276.
[22]BITTREMIEUX M,BULTYNCK G.p53and Ca2+signaling from the endoplasmic reticulum:partners in anticancer therapies[J].Oncoscience,2015,2(3):233-238
[23]KOKAWA K,SHIKONE T,OTANI T,et al.Apoptosis and the expression of Bax and Bcl-2in squamous cell carcinoma and adenocarcinoma of the uterine cervix[J].Cancer,2015,85(8):1799-1809.
[2]栗志勇,张慧,张婷,等.丹皮酚对急性心肌梗死大鼠氧化应激和内皮型一氧化氮合酶信号通路的影响[J].新乡医学院学报,2017,34(5):356-360.
[3]刘一杰,薛永常.植物黄酮类化合物的研究进展[J].中国生物工程杂志,2016,36(9):81-86.
[4]PRZEMYSLAW J S,SANDRA M P,SZUMNY A,et al.Quality parameters,bio-compounds,antioxidant activity and sensory attributes of Spanish quinces(Cydonia oblonga,Miller)[J].Scientia Horticulturae,2014,165(3):163-170.
[5]周文婷,马虎,阿迪力·阿不都热合曼,等.榅桲总黄酮对自发性高血压大鼠心脏结构和功能的影响[J].中成药,2016,38(11):2313-2318.
[6]马晓娟,郭春雨,张莹,等.活血及活血解毒配伍对急性心肌梗死大鼠心肌损伤的保护作用[J].中华老年心脑血管病杂志,2015,17(9):960-963.
[7]周文婷,王雪飞,邬利娅·伊明,等.榅桲总黄酮对自发性高血压大鼠心肌肥厚的抑制作用及机制研究[J].中国药理学通报,2015,31(11):1540-1546.
[8]赵洋,李艳.阿托伐他汀对心肌梗死大鼠心肌损伤的保护作用及机制[J].安徽医药,2016,20(2):240-243.
[9]马丕勇.生长激素对缺血性心力衰竭大鼠心肌细胞凋亡的影响[D].吉林大学,2011.
[10]蒋易,燕艳丽,白建文.CCL21/CCR7对小鼠急性心肌梗死后炎症反应及梗死面积的影响[J].临床与病理杂志,2017,37(3):589-594.
[11]姚若花,杨星,曹洁.单核细胞趋化蛋白-1与动脉粥样硬化关系的研究现状与进展[J].西南军医,2015,17(4):419-421.
[12]刘中勇,李林,方家.真武汤对心力衰竭模型大鼠心室重构及心肌细胞凋亡、纤维化的影响[J].中医杂志,2017,58(14):1218-1223.
[13]刘新育,顾剑民,薛松.NF-κB与心肌缺血再灌注损伤的研究现状及展望[J].中华胸心血管外科杂志,2016,32(1):55-58.
[14]JING C,HONG J,JIAN Y,et al.Down-regulation of CREB-binding protein expression blocks thrombin-mediated endothelial activation by inhibiting acetylation of NF-κB[J].Int J Cardiol,2012,154(2):147-152.
[15]陈彭生.五味子乙素促进心肌梗死小鼠心功能的研究[D].南京:南京医科大学,2014.
[16]何辉霞,郑维银.P38MAPK信号通路在血管内皮细胞凋亡作用中的研究进展[J/CD].中华临床医师杂志(电子版),2013,7(20):9317-9319.
[17]张云,旦菊花,孙鑫,等.MAPKs信号通路在动脉粥样硬化发生发展中的调控作用[J].现代生物医学进展,2017,17(5):968-970.
[18]KIM E K,CHOI E J.Compromised MAPK signaling in human diseases:an update[J].Arch Toxicol,2015,89(6):867-82.
[19]呼雅丽.黄癸固体分散体通过抑制JNK保护心肌缺血再灌注损伤的作用机制研究[D].吉林大学,2015.
[20]张腾,张艳军,庄朋伟,等.瓜蒌对心梗后心衰大鼠心功能及心肌细胞凋亡的影响[J].中国药理学通报,2016,32(8):1183-1184.
[21]GUO C,YANG M,JING L,et al.Amorphous silica nanoparticles trigger vascular endothelial cell injury through apoptosis and autophagy via reactive oxygen species-mediated MAPK/Bcl-2and PI3K/Akt/mTOR signaling[J].Int J Nanomedicine,2016,11:5257-5276.
[22]BITTREMIEUX M,BULTYNCK G.p53and Ca2+signaling from the endoplasmic reticulum:partners in anticancer therapies[J].Oncoscience,2015,2(3):233-238
[23]KOKAWA K,SHIKONE T,OTANI T,et al.Apoptosis and the expression of Bax and Bcl-2in squamous cell carcinoma and adenocarcinoma of the uterine cervix[J].Cancer,2015,85(8):1799-1809.