针灸预处理对心肌缺血大鼠预防性保护作用及机制研究
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摘要
目的:揭示针灸预处理的预防性心肌保护作用机制,为缺血性心脏病的防治开辟新途径,提供新的更加便捷安全有效方法。
方法:80只健康Wistar雄性大鼠,随机分为10组:正常对照组、假手术组、缺血再灌注组、缺血预处理组、手针预处理组、电针预处理组、艾灸预处理组,后三组又分为日1次,日2次。HE染色观察组织形态学变化、电镜观察组织超微结构、原位末端标记法检测心肌细胞凋亡、免疫组化和原位杂交检测Bcl-2mRNA、BaxmRNA、HSP70mRNA和蛋白表达、黄嘌呤氧化酶法测定SOD活性、硫代巴比妥酸法测MDA含量,比较各组梗塞范围。
结果:①针灸预处理组同缺血预处理组一样,能明显减少缺血后心肌细胞凋亡数目,与心肌缺血再灌注组比较具有显著性差异;②针灸预处理组Bcl-2、HSP70蛋白和基因表达明显增多,Bax蛋白和基因表达明显减少,与缺血再灌注组相比,差异显著;③针灸预处理后SOD活性明显增强;MDA含量显著下降,与心肌缺血再灌注组比较具有显著性差异;④光镜电镜观察针灸预处理组更有效减轻心肌损伤及梗塞范围;⑤针灸预处理日2次组比日1次组更有效。
结论:①针灸预处理可以延缓或者阻断试验性心肌缺血的发展进程,并且具有提高心肌组织在缺血时对一系列代谢紊乱的耐受能力,增加心肌细胞的存活率,改善预后的作用;②针灸预处理可以抑制心肌细胞凋亡;③针灸预处理可通过上调细胞凋亡基因Bcl-2mRNA,HSP70 mRNA和蛋白表达,下调Bax mRNA和蛋白表达,从而抑制缺血心肌组织细胞凋亡;④针灸预处理日2次组更有效预防心肌损伤;⑤针灸预处理可通过内源性抗氧化物质的增加对缺血缺氧心肌组织起保护作用。
Object:To reveal the mechanism of neuroprotective effects of acupuncture and moxibustion preconditioning,to break a new path for prevention and cure of ischemic heart disease.
Measure:60 rats were divided into ten groups randomly in the experiment: normal group,sham-operation group,ischemic group,ischemic preconditioning group,acupuncture preconditioning group,electric acupuncture preconditioning group, moxibustion preconditioning group,the last three groups are divided into once and twice.HE staining to observe change of histomorphology,electron microscope to observe ultrastructure,TUNEL method to observe apoptosis state, immunohistochemistry and ISH technique to detect Bcl-2mRNA、BaxmRNA、HSP70mRNA and protein expression,xanthine oxidase method to detect the active of SOD,Thibabituric Acid to dect the amount of MDA,comparison to detect infarction content.
Result:①Acupuncture and moxibustion preconditioning which is the same as ischemie preconditioning group can decrease cell apoptosis as ischemic preconditioning which showed remarkable difference compared with ischemic group.②Bcl-2mRNA、HSP70mRNA and protein expression increase significantly and BaxmRNA and protein expression reduce after acupuncture and moxibustion preconditioning.It was remarkable difference compared with ischemic group.③The active of SOD increase significantly after acupuncture and moxibustion preconditioning and the amount of MDA decrease significantly which showed remarkable difference compared with ischemic group.④infarction content and hurt of heart tissue is decreased after acupuncture and moxibustion preconditioning which showed remarkable difference compared with ischemic group.⑤acupuncture and moxibustion preconditioning twice group is better than acupuncture and moxibustion preconditioning once group.
Conclusion:①Acupuncture and moxibustion preconditioning can delay or abolish the further advance of ischemic cardiac muscle and increase the tolerance of ischemic heart tissue about a series of metabolic disturbances,increase the survival rate of cardiac cell,improve the outlook.②Acupuncture and moxibustion preconditioning can prevent cell apoptosis in heart.③Acupuncture and moxibustion preconditioning can prevent cell apoptosis in ischemic heart tissue by increasing Bcl-2mRNA,HSP70mRNA and protein expression,reducing BaxmRNA and protein expression.④Acupuncture and moxibustion preconditioning group twice prevents cardiac muscle from damage better than once.⑤Acupuncture and moxibustion preconditioning protects ischemic heart tissue by increasing the endogenic antioxidant.
方法:80只健康Wistar雄性大鼠,随机分为10组:正常对照组、假手术组、缺血再灌注组、缺血预处理组、手针预处理组、电针预处理组、艾灸预处理组,后三组又分为日1次,日2次。HE染色观察组织形态学变化、电镜观察组织超微结构、原位末端标记法检测心肌细胞凋亡、免疫组化和原位杂交检测Bcl-2mRNA、BaxmRNA、HSP70mRNA和蛋白表达、黄嘌呤氧化酶法测定SOD活性、硫代巴比妥酸法测MDA含量,比较各组梗塞范围。
结果:①针灸预处理组同缺血预处理组一样,能明显减少缺血后心肌细胞凋亡数目,与心肌缺血再灌注组比较具有显著性差异;②针灸预处理组Bcl-2、HSP70蛋白和基因表达明显增多,Bax蛋白和基因表达明显减少,与缺血再灌注组相比,差异显著;③针灸预处理后SOD活性明显增强;MDA含量显著下降,与心肌缺血再灌注组比较具有显著性差异;④光镜电镜观察针灸预处理组更有效减轻心肌损伤及梗塞范围;⑤针灸预处理日2次组比日1次组更有效。
结论:①针灸预处理可以延缓或者阻断试验性心肌缺血的发展进程,并且具有提高心肌组织在缺血时对一系列代谢紊乱的耐受能力,增加心肌细胞的存活率,改善预后的作用;②针灸预处理可以抑制心肌细胞凋亡;③针灸预处理可通过上调细胞凋亡基因Bcl-2mRNA,HSP70 mRNA和蛋白表达,下调Bax mRNA和蛋白表达,从而抑制缺血心肌组织细胞凋亡;④针灸预处理日2次组更有效预防心肌损伤;⑤针灸预处理可通过内源性抗氧化物质的增加对缺血缺氧心肌组织起保护作用。
Object:To reveal the mechanism of neuroprotective effects of acupuncture and moxibustion preconditioning,to break a new path for prevention and cure of ischemic heart disease.
Measure:60 rats were divided into ten groups randomly in the experiment: normal group,sham-operation group,ischemic group,ischemic preconditioning group,acupuncture preconditioning group,electric acupuncture preconditioning group, moxibustion preconditioning group,the last three groups are divided into once and twice.HE staining to observe change of histomorphology,electron microscope to observe ultrastructure,TUNEL method to observe apoptosis state, immunohistochemistry and ISH technique to detect Bcl-2mRNA、BaxmRNA、HSP70mRNA and protein expression,xanthine oxidase method to detect the active of SOD,Thibabituric Acid to dect the amount of MDA,comparison to detect infarction content.
Result:①Acupuncture and moxibustion preconditioning which is the same as ischemie preconditioning group can decrease cell apoptosis as ischemic preconditioning which showed remarkable difference compared with ischemic group.②Bcl-2mRNA、HSP70mRNA and protein expression increase significantly and BaxmRNA and protein expression reduce after acupuncture and moxibustion preconditioning.It was remarkable difference compared with ischemic group.③The active of SOD increase significantly after acupuncture and moxibustion preconditioning and the amount of MDA decrease significantly which showed remarkable difference compared with ischemic group.④infarction content and hurt of heart tissue is decreased after acupuncture and moxibustion preconditioning which showed remarkable difference compared with ischemic group.⑤acupuncture and moxibustion preconditioning twice group is better than acupuncture and moxibustion preconditioning once group.
Conclusion:①Acupuncture and moxibustion preconditioning can delay or abolish the further advance of ischemic cardiac muscle and increase the tolerance of ischemic heart tissue about a series of metabolic disturbances,increase the survival rate of cardiac cell,improve the outlook.②Acupuncture and moxibustion preconditioning can prevent cell apoptosis in heart.③Acupuncture and moxibustion preconditioning can prevent cell apoptosis in ischemic heart tissue by increasing Bcl-2mRNA,HSP70mRNA and protein expression,reducing BaxmRNA and protein expression.④Acupuncture and moxibustion preconditioning group twice prevents cardiac muscle from damage better than once.⑤Acupuncture and moxibustion preconditioning protects ischemic heart tissue by increasing the endogenic antioxidant.
引文
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