苦瓜总皂苷对心力衰竭大鼠心肌凋亡的影响
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摘要
目的探讨苦瓜总皂苷对心力衰竭大鼠心功能和心肌凋亡蛋白Caspase-8、Bcl-2表达的影响。方法取清洁级雄性SD大鼠60只,适应性喂养14d后按照随机数字表法分为正常组、模型组及苦瓜总皂苷低剂量、中剂量和高剂量组,每组12只。苦瓜总皂苷低剂量、中剂量和高剂量治疗组分别给予苦瓜总皂苷10mg/(kg·d)、20mg/(kg·d)、40mg/(kg·d)皮下注射,正常组和模型组分别给予0.5mL/(kg·d)生理盐水皮下注射。全程记录大鼠精神活动状态、毛色以及饮食等一般情况,治疗8周后处死大鼠,检测心功能相关指标以及炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平,苏木精-伊红染色(HE染色)检测心肌组织病理学改变;免疫组化、实时荧光定量聚合酶链式反应(qRT-PCR)、Western印迹检测心肌组织中Caspase-8和Bcl-2相关蛋白。结果正常组大鼠精神良好,饮食正常;模型组有明显精神萎靡,饮食量较低;苦瓜总皂苷低、中、高剂量治疗组较模型组上述症状明显减轻。与正常组比较,模型组平均心室舒张压(LVDP)明显升高(P <0.05),平均左室峰压(LVSP)、心室内压最大上升速率(+dp/dtmax)及平均等容舒张期室内压最大下降速率(-dp/dtmax)明显降低(P <0.05);与模型组比较,苦瓜总皂苷中、高剂量组LVDP明显降低(P <0.05),LVSP、+dp/dtmax和-dp/dtmax明显升高(P <0.05)。与正常组比较,模型组TNF-α、IL-6明显增加(P <0.05);与模型组比较,苦瓜总皂苷低、中、高剂量治疗组TNF-α、IL-6明显降低(P <0.05);HE染色可见正常组大鼠心肌细胞排列整齐,而模型组有明显心肌细胞排列紊乱,苦瓜总皂苷低、中、高剂量组心肌组织较模型组均有不同程度的改善。与正常组比较,模型组大鼠心肌组织Bcl-2蛋白表达明显下降(P <0.05),Caspase-8蛋白表达明显升高(P <0.05);与模型组比较,苦瓜总皂苷中、高剂量组Bcl-2蛋白表达均升高(P <0.05),Caspase-8蛋白表达则明显降低(P <0.05)。结论苦瓜总皂苷可以明显改善慢性心力衰竭大鼠心功能,降低炎性因子TNF-α、IL-6的表达,可能与其下调Caspase-8、上调Bcl-2表达有关。
Objective To investigate the effects of Momordica charantia saponins(MCS)on the cardiac function and the expression of Caspase-8 and Bcl-2 in rats with heart failure.Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into the normal group,model group,MCS low dose group,MCS medium dose group,and MCS high dose group,12 rats in each group.The chronic heart failure model was established by the treatment of isoproterenol hydrochloride.After successful modeling,the rats were administered with subcutaneous injection of MCS 10 mg/(kg·d),20 mg/(kg·d),and 40 mg/(kg·d)in MCS low dose group,MCS medium dose group,and MCS high dose group,respectively.the rats in the normal group and the model group were given with subcutaneous injection of 0.5 mL/(kg·d)saline.The rats were sacrificed after 8 weeks of treatment.The cardiac function related indicators,and inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)levels were detected.The myocardial histopathological changes were detected by Hematoxylin-Eosin(HE)staining.The expressions of Caspase-8 and Bcl-2 related proteins in myocardium were detected by immunohistochemistry,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),western blot,and real-time fluorescent PCR.Results The rats in the normal group had a good mental and normal diet,while the rats in model group had obvious apathy,compared with the model group,the above symptoms were significantly reduced.Compared with the normal group,the left ventricular pressure peak(LVSP),+dp/dtmax and-dp/dtmax were significantly decreased in the model group(P <0.05).Compared with the model group,left ventricular diastolic pressure(LVDP)was significantly decreased(P <0.05),LVSP,+dp/dtmax and-dp/dtmax were significantly increased in MCS low dose group,MCS medium dose group and MCS high dose group(P <0.05).Compared with the normal group,TNF-αand IL-6 in the model group were significantly increased after treatment(P <0.05).Compared with the model group,TNF-α,and IL-6 were significantly decreased in MCS low dose group,MCS medium dose group and MCS high dose group after treatment(P <0.05).HE staining showed that the cardiomyocytes were neatly arranged in the normal group,and had obvious disorder in the model group.The myocardial tissue in MCS low dose group,MCS medium dose group and MCS high dose group had different degrees of improvement compared with the model group.Compared with the normal group,the expression of Bcl-2 protein in the model group was significantly lower than that in the normal group(P <0.05),and the expression of Caspase-8 protein in the model group was significantly higher than that in the normal group(P <0.05).Compared with the model group,the expression of Bcl-2 protein in MCS groups were increased(P <0.05),Caspase-8 expression was significantly decreased(P <0.05).Conclusion MCS can significantly improve the cardiac function and decrease the expression of inflammatory factors TNF-αand IL-6 in rats with chronic heart failure,which may be related to down-regulating Caspase-8 and up-regulating Bcl-2 expression.
Objective To investigate the effects of Momordica charantia saponins(MCS)on the cardiac function and the expression of Caspase-8 and Bcl-2 in rats with heart failure.Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into the normal group,model group,MCS low dose group,MCS medium dose group,and MCS high dose group,12 rats in each group.The chronic heart failure model was established by the treatment of isoproterenol hydrochloride.After successful modeling,the rats were administered with subcutaneous injection of MCS 10 mg/(kg·d),20 mg/(kg·d),and 40 mg/(kg·d)in MCS low dose group,MCS medium dose group,and MCS high dose group,respectively.the rats in the normal group and the model group were given with subcutaneous injection of 0.5 mL/(kg·d)saline.The rats were sacrificed after 8 weeks of treatment.The cardiac function related indicators,and inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)levels were detected.The myocardial histopathological changes were detected by Hematoxylin-Eosin(HE)staining.The expressions of Caspase-8 and Bcl-2 related proteins in myocardium were detected by immunohistochemistry,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),western blot,and real-time fluorescent PCR.Results The rats in the normal group had a good mental and normal diet,while the rats in model group had obvious apathy,compared with the model group,the above symptoms were significantly reduced.Compared with the normal group,the left ventricular pressure peak(LVSP),+dp/dtmax and-dp/dtmax were significantly decreased in the model group(P <0.05).Compared with the model group,left ventricular diastolic pressure(LVDP)was significantly decreased(P <0.05),LVSP,+dp/dtmax and-dp/dtmax were significantly increased in MCS low dose group,MCS medium dose group and MCS high dose group(P <0.05).Compared with the normal group,TNF-αand IL-6 in the model group were significantly increased after treatment(P <0.05).Compared with the model group,TNF-α,and IL-6 were significantly decreased in MCS low dose group,MCS medium dose group and MCS high dose group after treatment(P <0.05).HE staining showed that the cardiomyocytes were neatly arranged in the normal group,and had obvious disorder in the model group.The myocardial tissue in MCS low dose group,MCS medium dose group and MCS high dose group had different degrees of improvement compared with the model group.Compared with the normal group,the expression of Bcl-2 protein in the model group was significantly lower than that in the normal group(P <0.05),and the expression of Caspase-8 protein in the model group was significantly higher than that in the normal group(P <0.05).Compared with the model group,the expression of Bcl-2 protein in MCS groups were increased(P <0.05),Caspase-8 expression was significantly decreased(P <0.05).Conclusion MCS can significantly improve the cardiac function and decrease the expression of inflammatory factors TNF-αand IL-6 in rats with chronic heart failure,which may be related to down-regulating Caspase-8 and up-regulating Bcl-2 expression.
引文
[1]刘晓帅,王果,汪林,等.桂枝茯苓丸抗大鼠慢性心衰的实验研究[J].西南民族大学学报(自然科学版),2017,43(4):378-385.
[2]方焕松,刘友章,邱俊,等.四君子汤对慢性心衰大鼠心肌组织蛋白质组学影响的研究[J].中国实验方剂学杂志,2017(5):152-156.
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[4]陈珏,於悦,高想,等.白藜芦醇调控大鼠心力衰竭模型氧化应激相关基因的作用[J].中西医结合心脑血管病杂志,2016,14(7):704-706.
[5]刘茂,陈剑,姚婕榕,等.雷公藤内酯上调PTEN信号通路减轻异丙肾上腺素诱导的慢性心力衰竭大鼠心室重构[J].中山大学学报(医学科学版),2017,38(1):29-35.
[6]刘紫燕,朱定君,王丽媛,等.苦瓜总皂苷对2型糖尿病大鼠心肌脂联素受体1表达及心功能的影响[J].长春中医药大学学报,2017,33(1):40-43.
[7]赵英强,孙兰军,徐强,等.慢性充血性心力衰竭大鼠动物模型制备[J].天津中医药,2004,21(4):318-321.
[8]董安平,李晓峰,马爱群.大鼠慢性容量超负荷心力衰竭模型的建立[J].医学争鸣,2003,24(18):1682-1684.
[9]钟泽,罗秀英,刘桂林,等.SD大鼠3种慢性心力衰竭模型的建立及心功能比较[J].心电与循环,2014,33(4):307-310.
[10]杨喆,柴洪佳,王大伟.阳虚型慢性心力衰竭SD大鼠模型的制备及指标判定[J].中西医结合心脑血管病杂志,2016,14(14):1593-1596.
[11]史文静,周华,戎靖枫.温补心阳和肾阳方对心力衰竭大鼠心功能和心肌磷酸腺苷含量的影响[J].中西医结合心脑血管病杂志,2012,10(9):1083-1085.
[12]张锦,赵翠翠,陆景坤,等.5种黄酮类化合物对凋亡心肌细胞Bcl-2家族蛋白表达的影响[J].中华地方病学杂志,2008,27(2):149-151.
[13]王青,蔡剑峰,郑婷婷,等.重楼总皂苷对A549细胞凋亡及Caspase 3、Bcl-2蛋白表达的影响[J].中华中医药学刊,2017(7):1708-1710.
[14]袁芳,李艳青,滕旭,等.慢性间歇性低压低氧抑制线粒体途径介导的代谢综合征大鼠心肌组织细胞凋亡[J].中国药理学通报,2015,31(8):1131-1136.
[15]OCHS K,KAINA B.Apoptosis induced by DNA damage O-methylguanine is Bcl-2 and caspase-9/3 regulated and Fas/caspase-8independent[J].Cancer Research,2000,60(20):5815.
[16]柴瑞华,肖春莹,关健,等.苦瓜总皂苷降血糖作用的研究[J].中草药,2007,39(2):746-747.
[17]莫灼康,香富强.苦瓜总皂苷对高血脂模型小鼠的影响[J].中国社区医师:医学专业,2012,14(12):8.
[18]朱定君,殷旭光,刘紫燕,等.苦瓜总皂苷对Ⅱ型糖尿病大鼠心肌细胞凋亡因子Fas和FasL表达的影响[J].西部医学,2016,28(10):1360-1364.
[19]朱定君,刘紫燕,殷旭光,等.苦瓜总皂苷对2型糖尿病大鼠心脏组织中MMP-9和HO-1表达影响[J].辽宁中医药大学学报,2017(1):48-51.
[2]方焕松,刘友章,邱俊,等.四君子汤对慢性心衰大鼠心肌组织蛋白质组学影响的研究[J].中国实验方剂学杂志,2017(5):152-156.
[3]黄峻.中国心力衰竭流行病学特点和防治策略[J].中华心脏与心律电子杂志,2015,3(2):2-3.
[4]陈珏,於悦,高想,等.白藜芦醇调控大鼠心力衰竭模型氧化应激相关基因的作用[J].中西医结合心脑血管病杂志,2016,14(7):704-706.
[5]刘茂,陈剑,姚婕榕,等.雷公藤内酯上调PTEN信号通路减轻异丙肾上腺素诱导的慢性心力衰竭大鼠心室重构[J].中山大学学报(医学科学版),2017,38(1):29-35.
[6]刘紫燕,朱定君,王丽媛,等.苦瓜总皂苷对2型糖尿病大鼠心肌脂联素受体1表达及心功能的影响[J].长春中医药大学学报,2017,33(1):40-43.
[7]赵英强,孙兰军,徐强,等.慢性充血性心力衰竭大鼠动物模型制备[J].天津中医药,2004,21(4):318-321.
[8]董安平,李晓峰,马爱群.大鼠慢性容量超负荷心力衰竭模型的建立[J].医学争鸣,2003,24(18):1682-1684.
[9]钟泽,罗秀英,刘桂林,等.SD大鼠3种慢性心力衰竭模型的建立及心功能比较[J].心电与循环,2014,33(4):307-310.
[10]杨喆,柴洪佳,王大伟.阳虚型慢性心力衰竭SD大鼠模型的制备及指标判定[J].中西医结合心脑血管病杂志,2016,14(14):1593-1596.
[11]史文静,周华,戎靖枫.温补心阳和肾阳方对心力衰竭大鼠心功能和心肌磷酸腺苷含量的影响[J].中西医结合心脑血管病杂志,2012,10(9):1083-1085.
[12]张锦,赵翠翠,陆景坤,等.5种黄酮类化合物对凋亡心肌细胞Bcl-2家族蛋白表达的影响[J].中华地方病学杂志,2008,27(2):149-151.
[13]王青,蔡剑峰,郑婷婷,等.重楼总皂苷对A549细胞凋亡及Caspase 3、Bcl-2蛋白表达的影响[J].中华中医药学刊,2017(7):1708-1710.
[14]袁芳,李艳青,滕旭,等.慢性间歇性低压低氧抑制线粒体途径介导的代谢综合征大鼠心肌组织细胞凋亡[J].中国药理学通报,2015,31(8):1131-1136.
[15]OCHS K,KAINA B.Apoptosis induced by DNA damage O-methylguanine is Bcl-2 and caspase-9/3 regulated and Fas/caspase-8independent[J].Cancer Research,2000,60(20):5815.
[16]柴瑞华,肖春莹,关健,等.苦瓜总皂苷降血糖作用的研究[J].中草药,2007,39(2):746-747.
[17]莫灼康,香富强.苦瓜总皂苷对高血脂模型小鼠的影响[J].中国社区医师:医学专业,2012,14(12):8.
[18]朱定君,殷旭光,刘紫燕,等.苦瓜总皂苷对Ⅱ型糖尿病大鼠心肌细胞凋亡因子Fas和FasL表达的影响[J].西部医学,2016,28(10):1360-1364.
[19]朱定君,刘紫燕,殷旭光,等.苦瓜总皂苷对2型糖尿病大鼠心脏组织中MMP-9和HO-1表达影响[J].辽宁中医药大学学报,2017(1):48-51.