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复方甲亢片对甲亢大鼠合并心脏病变的影响及其作用机理研究
【作者】 方居正;
【导师】 陈如泉;
【作者基本信息】 湖北中医学院 , 中医内科学, 2006, 博士
【摘要】 目的 通过采用灌胃法诱导建立甲亢大鼠合并心脏病变的模型,运用生化、电镜、高效液相色谱法、逆转录多聚酶链法(RT-PCR),检测高甲状腺素水平下,大鼠心肌结构、超微结构、生理功能、激素水平、细胞因子、病理基因,确定造模的可靠性。使用复方甲亢片、纯中药、甲巯咪唑等进行干预,比较三种药物的疗效,从而得出结论。探索复方甲亢片治疗甲亢大鼠合并心脏病变的机理,以期为中西药结合治疗甲亢性心脏病提供理论依据。 方法 清洁级SD大鼠60只,体重约250±15g,雌雄各半。随机分为五组,正常组、模型组、纯中药组、复方甲亢片组、甲巯咪唑组。每组十二只。正常组每天上午给以2ml蒸馏水喂服,其余四组均造模。造模方法,左旋甲状腺素钠片混悬液上午喂服,每日1mg/kg,每天一次。连续十七天。自造模第十七天始,正常组仍继续每天上午给以2ml蒸馏水喂服,其余四组均继续造模,继续左旋甲状腺素钠片混悬液上午喂服,每日1mg/kg,每天一次。此外下午加喂正常组与模型组大鼠每日灌2ml蒸馏水,复方甲亢片组每日下午加灌复方甲亢片混悬液2ml(相当于生药8g/kg),纯中药组每日下午加灌服纯中药液2ml(相当于生药8g/kg)。甲巯咪唑组每日下午加灌服甲巯咪唑液2ml(相当于12mg/kg)。共连续五十天实验结束。实验结束后,每大鼠以20%乌拉坦麻醉(剂量0.5ml/100g体重),腹腔注射麻醉。以生理二道仪记录心电图,眼眶取血,心脏称重,检测血浆BNP、血清TT3、TT4,心肌匀浆测羟普氨酸和Na+-K+-ATP酶。心肌HE染色进行病理切片检测,超薄切片进行透射电镜检测。免疫组化检测MMP2、bFGF、TNF-α因子。逆转录-多聚酶链反应法(RT-PCR)检测肌球蛋白重链的表型(α-MHC和β-MHC)。 结果 1、复方甲亢片能比较明显改善甲亢大鼠合并的心脏疾病症状,提高生存率。 2、经检测大鼠血清TT3、TT4水平,检测大鼠心率及心律,显示模型组大鼠血清TT3、TT4水平、心率和心律失常发生率显著升高。心脏指数为最大。正常组大鼠血清TT3、TT4水平、心率和心律失常发生率最低,心脏指数为最低。经药物治疗后所有用药组血清TT3、TT4水平都有不同程度降低,心率、心律失常发生率、心脏指数也有不同程度下降。以复方甲亢片组血清TT3、TT4水平、心率、心律失常发生率、心脏指数降低为最明显。从统计结果看,正常组与模型组相比P<0.01,有极显著差异。经治疗后,纯中药组和甲巯咪唑组与模型组相比P<0.05,均有显著差异,说明两种药物均有效。复方甲亢片组与模型组相比P<0.01,有极显著差异,与甲巯咪唑和纯中药相比P<0.05,均有显著差异。说明复方甲亢片有效,并且效果优于纯中药和甲巯咪唑。 3、光镜下检测,HE染色显示正常组心肌细胞排列整齐,横纹清楚,间质较少,几乎看不到成纤维细胞。与正常组相比可见到模型组心
【Abstract】 Objective The model that the rats are filled by thyroxin are examined , their cardic muscle structure , ultrastructure, physiology structure, hormone level, cell gene and pathology gene are inspected through biochemistry, electron microscope , super liquid chromatogram and RT-PCR .the reliability of model are confirmed. And then they are cured by Fufangjiakang tablet, pure traditional Chinese drug and thiamazole, the three kinds of medicine are compared and a conclusion is drawn. In order to provide theroy testimony of curing Thyroxin cardiopathy with combination of TCM with western method, we grope for principle that Thyroxin cardiopathy is cured by Fufangjiakangpian. Method Sixty clean rats(male and female are half and half) were classified into five groups at random , they were control group, model group , pure traditional Chinese drug group, thiamazole group and fufangjiakang group. Every group had twelve rats. Control group rats were filled by 2ml distilled water every moyning, the other group rats were filled by L-thyroxin every morning.lmg/Kg every morning. After seventeen days, control group rats and model group rats were filled by 2 ml distilled water in addition every afternoon, pure traditional Chinese drug group rats were filled by pure traditional Chinese drug( dose 8g/kg)