氧化苦参碱改善病毒性心肌炎小鼠心室重构的作用研究
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摘要
目的观察氧化苦参碱对病毒性心肌炎所致心力衰竭小鼠心室重构、心肌能量代谢的影响。方法将50只雄性BALB/c小鼠随机分为对照组、模型组(腹腔无菌注射0.1 ml 1×10~(-9) TCID50柯萨奇B3病毒),以及低、中、高剂量组(在模型组基础上分别皮下注射氧化苦参碱13、26和52 mg/kg)。连续给药14 d后,采用高频超声心动图检测各组小鼠心功能;HE染色、透射电镜观察左心室肌超微结构;TUNEL法检测各组小鼠左心室心肌细胞凋亡情况;ELISA法检测左室心肌细胞游离脂肪酸(FFA)、三磷酸腺苷(ATP)、腺苷酸(AMP)、乳酸(LAC)、Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)含量,Wsetern blotting检测各组小鼠左室心肌细胞葡萄糖调节蛋白78(GRP78)、C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白(CHOP)、骨膜蛋白、钙联蛋白的表达。结果模型组较对照组左室舒张末期内径扩大,左室射血分数和左室短轴缩短率减小(P <0.05);左室心肌细胞核排列紊乱、线粒体空泡化、肌丝断裂,出现大量病毒,心肌细胞凋亡数量增多(P <0.05);FFA、LAC、ColⅠ、ColⅢ、GRP78、CHOP、骨膜蛋白表达升高(P <0.05);ATP/AMP比值及钙联蛋白表达降低(P<0.05)。低、中、高剂量组较模型组左心室心肌细胞病理变化及左室重构指标改善(P <0.05);FFA、LAC、ColⅠ、ColⅢ、GRP78、CHOP、骨膜蛋白表达降低(P <0.05);ATP/AMP比值及钙联蛋白表达升高(P<0.05)。结论氧化苦参碱可通过保护线粒体,改善心肌能量代谢,减少心肌细胞凋亡,逆转心室重构,起到改善病毒性心肌炎小鼠心力衰竭的作用。
Objective To observe the effect of Oxymatrine on ventricular remodeling and myocardial energy metabolism in mice with congestive heart failure caused by viral myocarditis. Methods Fifty male BALB/c mice were randomly divided into a control group, a model group(intraperitoneal injection of 0.1 ml 1×10~(-9) TCID50 Coxsackie B3 virus), a low-dose group, a medium-dose group and a high-dose group(subcutaneous injection of Oxymatrine 13, 26 and 52 mg/kg into the model mice). After continuous administration for 14 d, cardiac function of each group was detected by high frequency echocardiography; structures and ultrastructures of left ventricular muscle were observed by HE staining and transmission electron microscopy; apoptosis of left ventricular cardiomyocytes was detected by TUNEL method; the levels of free fatty acids(FFA), adenosine triphosphate(ATP), adenosine monophosphate(AMP), lactic acid(LA), type Ⅰ collagen(Col Ⅰ) and type Ⅲ collagen(Col Ⅲ) in the left ventricular cardiomyocytes were detected by ELISA; Western blot was used to detect the expression of glucose-regulated protein 78(GRP78) and C/EBP cyclic adenosine monophosphate response element binding transcription factor homologous protein(CHOP), periostin and calnexin in the left ventricular cardiomyocytes of mice. Results In the model group, LVEDD was enlarged, LVEF and FS were decreased(P < 0.05); the nuclei of the left ventricular cardiomyocytes were disordered, the mitochondria were vacuolated, the myofilaments were broken and the virus appeared, the number of myocardial cells was increased(P < 0.05); the expressions of FFA, LA, Col Ⅰ, Col Ⅲ, GRP78, CHOP and periostin were increased(P < 0.05), the ratio of ATP/AMP and the expression of calnexin were decreased(P < 0.05). The pathological changes and left ventricular remodeling indexes of left ventricular cardiomyocytes in the lowdose, medium-dose and high-dose groups were improved(P < 0.05), the expressions of FFA, LA, Col Ⅰ, Col Ⅲ, GRP78, CHOP and periostin were decreased(P < 0.05), the ATP/AMP ratio and calnexin expression were increased(P < 0.05). Conclusions Oxymatrine can ameliorate the heart failure in mice with viral myocarditis by protecting mitochondria, improving myocardial energy metabolism, reducing myocardial cell apoptosis and reversing ventricular remodeling.
Objective To observe the effect of Oxymatrine on ventricular remodeling and myocardial energy metabolism in mice with congestive heart failure caused by viral myocarditis. Methods Fifty male BALB/c mice were randomly divided into a control group, a model group(intraperitoneal injection of 0.1 ml 1×10~(-9) TCID50 Coxsackie B3 virus), a low-dose group, a medium-dose group and a high-dose group(subcutaneous injection of Oxymatrine 13, 26 and 52 mg/kg into the model mice). After continuous administration for 14 d, cardiac function of each group was detected by high frequency echocardiography; structures and ultrastructures of left ventricular muscle were observed by HE staining and transmission electron microscopy; apoptosis of left ventricular cardiomyocytes was detected by TUNEL method; the levels of free fatty acids(FFA), adenosine triphosphate(ATP), adenosine monophosphate(AMP), lactic acid(LA), type Ⅰ collagen(Col Ⅰ) and type Ⅲ collagen(Col Ⅲ) in the left ventricular cardiomyocytes were detected by ELISA; Western blot was used to detect the expression of glucose-regulated protein 78(GRP78) and C/EBP cyclic adenosine monophosphate response element binding transcription factor homologous protein(CHOP), periostin and calnexin in the left ventricular cardiomyocytes of mice. Results In the model group, LVEDD was enlarged, LVEF and FS were decreased(P < 0.05); the nuclei of the left ventricular cardiomyocytes were disordered, the mitochondria were vacuolated, the myofilaments were broken and the virus appeared, the number of myocardial cells was increased(P < 0.05); the expressions of FFA, LA, Col Ⅰ, Col Ⅲ, GRP78, CHOP and periostin were increased(P < 0.05), the ratio of ATP/AMP and the expression of calnexin were decreased(P < 0.05). The pathological changes and left ventricular remodeling indexes of left ventricular cardiomyocytes in the lowdose, medium-dose and high-dose groups were improved(P < 0.05), the expressions of FFA, LA, Col Ⅰ, Col Ⅲ, GRP78, CHOP and periostin were decreased(P < 0.05), the ATP/AMP ratio and calnexin expression were increased(P < 0.05). Conclusions Oxymatrine can ameliorate the heart failure in mice with viral myocarditis by protecting mitochondria, improving myocardial energy metabolism, reducing myocardial cell apoptosis and reversing ventricular remodeling.
引文
[1]刘磊,王洪军,辛晴,等.病毒性心肌炎所致小鼠心力衰竭心肌组织内质网应激相关的凋亡关系的研究[J].中国应用生理学杂志,2014,30(5):461-464.
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[7]刘阳,刘念,刘文.氧化苦参碱联合5-FU对人胃癌SGC-7901细胞生长的协同抑制作用及其机制研究[J].中国免疫学杂志,2016,32(12):1781-1789.
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[9]桑秀秀,王睿林,韩延忠,等.养护苦参碱通过免疫调节保护刀豆蛋白A诱导的小鼠免疫性肝损伤[J].中国实验方剂学杂志,2017,23(6):125-130.
[10]易云,吴琴,黄莉萍,等.氧化苦参碱对高糖毒性下血管内皮细胞的保护作用[J].中国药理学通报,2015,31(4):499-503.
[11]杨志伟,周娅,曹秀琴.苦豆子生物碱体外抗柯萨奇B3病毒的作用[J].四川中医,2003,21(3):14-16.
[12]只达石,黄慧玲,张国斌,等.体温对颅脑创伤患者细胞间液葡萄糖和乳酸的影响[J].中国医学科学院学报,2006,28(2):262-266.
[13]张东莲,王洪新,梁灵君,等.人参多糖对腹主动脉缩窄大鼠心肌肥厚及心肌能量代谢的影响[J].中国现代应用药学,2013,30(6):571-575.
[14]SHIMAZAKI M,NAKAMURA K,KII I,et al.Periostin is essential for cardiac healing after acute myocardial infarction[J].J Exp Med,2008,205(2):295-303.
[15]邹平,吴丹,吴立玲,等.AngⅡ促进成年大鼠心脏成纤维细胞Periostin表达的信号通路研究[J].现代预防医学,2011,38(16):3265-3272.
[16]张峰.内质网分子伴侣Calnexin的研究进展[J].生物学通报,2008,43(8):7-10.
[17]ZHOU X M,XIN Q,WANG Y,et al.Total flavonoids of astragalus play a cardioprotective role in viral myocarditis[J].Acta Cardiol Sin,2016,32(1):81-88.
[2]张建明,朱锋,牟华明,等.缺血性心力衰竭患者血浆Periostin检测的临床意义[J].中国老年学杂志,2013,33(20):5114-5115.
[3]SMITH J L,MCBRIDE C M,NATARAI P S,et al.Traffickingdeficient hERG K+channels linked to long QT syndrome are regulated by a microtubule-dependent quality control compartment in the ER[J].Am J Physiol Cell Physiol,2011,301(1):C75-C85.
[4]汪洋,徐烨华,熊爱琴,等.氧化苦参碱抑制异丙肾上腺素诱导大鼠慢性心力衰竭及对ADMA代谢通路的影响[J].中国中药杂志,2014,39(3):471-477.
[5]陈碧涛,朱荣金,杨红宇,等.氧化苦参碱对病毒性心肌炎小鼠心肌细胞凋亡及相关因子的影响[J].中国现代应用药学,2016,33(3):162-165.
[6]张鸣号,王秀玉,何沿虹,等.氧化苦参碱对感染性休克大鼠心肌组织细胞因子的影响[J].中草药,2012,43(11):2242-2246.
[7]刘阳,刘念,刘文.氧化苦参碱联合5-FU对人胃癌SGC-7901细胞生长的协同抑制作用及其机制研究[J].中国免疫学杂志,2016,32(12):1781-1789.
[8]王伊林,赵雅君,杨洋,等.磷酸肌酸钠对慢病毒介导Calumenin蛋白沉默阿霉素损伤心肌细胞内质网应激信号通路的作用[J].临床心血管病杂志,2015,31(10):1119-1122.
[9]桑秀秀,王睿林,韩延忠,等.养护苦参碱通过免疫调节保护刀豆蛋白A诱导的小鼠免疫性肝损伤[J].中国实验方剂学杂志,2017,23(6):125-130.
[10]易云,吴琴,黄莉萍,等.氧化苦参碱对高糖毒性下血管内皮细胞的保护作用[J].中国药理学通报,2015,31(4):499-503.
[11]杨志伟,周娅,曹秀琴.苦豆子生物碱体外抗柯萨奇B3病毒的作用[J].四川中医,2003,21(3):14-16.
[12]只达石,黄慧玲,张国斌,等.体温对颅脑创伤患者细胞间液葡萄糖和乳酸的影响[J].中国医学科学院学报,2006,28(2):262-266.
[13]张东莲,王洪新,梁灵君,等.人参多糖对腹主动脉缩窄大鼠心肌肥厚及心肌能量代谢的影响[J].中国现代应用药学,2013,30(6):571-575.
[14]SHIMAZAKI M,NAKAMURA K,KII I,et al.Periostin is essential for cardiac healing after acute myocardial infarction[J].J Exp Med,2008,205(2):295-303.
[15]邹平,吴丹,吴立玲,等.AngⅡ促进成年大鼠心脏成纤维细胞Periostin表达的信号通路研究[J].现代预防医学,2011,38(16):3265-3272.
[16]张峰.内质网分子伴侣Calnexin的研究进展[J].生物学通报,2008,43(8):7-10.
[17]ZHOU X M,XIN Q,WANG Y,et al.Total flavonoids of astragalus play a cardioprotective role in viral myocarditis[J].Acta Cardiol Sin,2016,32(1):81-88.