连夏配方颗粒对心肌梗死后大鼠心脏自主神经重构的影响

发布时间：2018-05-16 21:09

本文选题：连夏配方颗粒 + NGF/TrKA/SH2B　；参考：《北京中医药大学》2015年博士论文

【摘要】：心肌梗死发生后,心肌梗死周边区的交感神经与迷走神经均会发生神经重构。而由此引发心电生理以及复极离散系列性、继发性的紊乱成为心肌梗死后致命性心律失常发生的关键性因素。而心肌梗死周围区则是心肌梗死发生后心肌组织修复、神经再生的活跃区域,与心肌梗死中心区域和正常区域相比更加显著。找到有效地解决心肌梗死后心脏自主神经重构的办法迫在眉睫。本论文将从文献综述、系统评价、实验研究三个方面对心肌梗死后心梗周边区心脏自主神经重构进行探讨,以期发现连夏配方颗粒在改善心肌梗死后心梗周边区心脏自主神经重构过程中的作用与机制。第一部分文献综述就近年来国内外对心肌梗死后心脏交感神经重构与心律失常的关系、神经生长因子与心血管疾病的相关性以及神经生长因子受体生理功能及其信号转导等研究进展进行了详细综述,介绍了目前国内外有关心肌梗死后心脏自主神经重构研究中的最新理论观点、工作进展情况以及未来研究发展趋势等,并初步讨论了目前研究中仍然存在、尚需解决的问题。第二部分系统评价目的系统评价黄连温胆汤治疗心律失常的疗效和安全性。方法全面收集黄连温胆汤治疗心律失常的随机对照试验(randomized controlled trial,RCT),根据纳入标准严格筛选,使用STATA11.0分析软件进行Meta荟萃分析。结果黄连温胆汤组可以明显改善心律失常的各结局指标,并且其疗效均高于对照组。二者综合疗效总有效率合并效应量为[RR=1.267,95%CI:1.157-1.386, P=0.000]、综合疗效显效率合并效应量为[RR=1.340,95%CI:1.111-1.616, P=0.002]、临床症状总有效率合并效应量为[RR=1.238,95%CI:1.015-1.510, P=0.035]、动态心电图室早总有效率合并效应量为[RR=1.256,95%CI:1.003-1.573, P=0.047]。结论黄连温胆汤能够明显改善心律失常的综合疗效总有效率与显效率、临床症状总有效率和动态心电图室早总有效率等相关指标,但是由于本次系统评价所纳入研究的方法质量不高和可能存在发表偏倚等不良因素,本次系统评价的结果与结论尚需要高质量、多中心的随机双盲临床对照试验来进一步证实。第三部分实验研究目的观察TH、AchE、NGF、TrKA、SH2B、Erkl/2的mRNA与TH、AchE、NGF、 TrKA、p-TrkA、SH2B、Erkl/2、 p-Erkl/2的蛋白表达水平变化在连夏配方颗粒干预心肌梗死后心梗周边区心脏自主神经重构中的作用。方法将105只健康雄性清洁级42-49日龄Wistar大鼠,按照随机数字表法随机分为正常组、假手术组、心梗模型组(模型组)、美托洛尔组(西药组)、连夏配方颗粒大、中、小剂量(189.00 mg/kg、94.50 mg/kg、47.25 mg/kg)组共计7个组,每组大鼠15只。正常组大鼠不做处理,美托洛尔组、中药大、中、小剂量组大鼠均结扎左冠状动脉前降支以建立急性心肌梗死大鼠模型,假手术组大鼠只穿线不结扎。于模型建立当天即开始灌胃,正常组、假手术与模型组大鼠予生理盐水灌胃,余各组均予相应实验药物。连续灌胃给药30天后麻醉大鼠,迅速于冰上获取大鼠心肌梗死周围区相同部位处足量心肌组织,Real Time PCR方法检测心肌组织中TH、AchE、NGF、TrKA、SH2B、Erkl/2的mRNA表达水平；Western Blot方法检测心肌组织中TH、AchE、NGF、TrKA、p-TrkA、 SH2B、 Erkl/2、p-Erkl/2的蛋白表达水平。结果①对TH mRNA的影响：与正常组、假手术组比较,模型组大鼠心梗周围区心肌组织TH mRNA表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50、47.25mg/kg组大鼠心梗周围区TH mRNA表达均明显降低(P0.05)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区THmRNA表达明显降低(P0.05)。②对AchE mRNA的影响：与正常组、假手术组比较,心梗模型组大鼠心梗周围区AchE mRNA表达显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组大鼠心梗周围区AchE mRNA表达未见明显降低(P0.05)；连夏配方颗粒189.00、94.50、47.25 mg/kg组大鼠心梗周围区AchE mRNA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区AchE mRNA表达显著降低(P0.01)；连夏配方颗粒94.50 mg/kg组大鼠心梗周围区AchE mRNA表达明显降低(P0.05)。③对NGF mRNA的影响：与正常组、假手术比较,心梗模型组大鼠心梗周围区NGF mRNA表达显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50、47.25 mg/kg组大鼠心梗周围区NGFmRNA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区NGF mRNA表达明显降低(P0.05)。④对TrKA mRNA的影响：与正常组、假手术组比较,心梗模型组大鼠心梗周围区TrKA mRNA表达显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50mg/kg组大鼠心梗周围区TrKA mRNA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00、94.50 mg/kg组大鼠心梗周围区TrKA mRNA表达均未见明显变化(P0.05)。⑤对SH2B mRNA的影响：与正常组、假手术组比较,心梗模型组大鼠心梗周围区SH2B mRNA表达显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50、47.25mg/kg组大鼠心梗周围区SH2B mRNA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg 组大鼠心梗周围区 SH2B mRNA表达明显降低(P0.05)。⑥对Erk1/2 mRNA的影响：与正常组、假手术组比较,心梗模型组大鼠心梗周围区Erk1/2 mRNA表达显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50mg/kg组大鼠心梗周围区Erkl/2 mRNA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00mg/kg组大鼠心梗周围区Erk1/2 mRNA表达显著降低(P0.01)。⑦对TH的影响：与正常组、假手术组比较,模型组大鼠心梗周围区TH表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组大鼠心梗周围区TH表达明显降低(P0.05)；连夏配方颗粒189.00 mg/kg组大鼠心梗周围区TH表达显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区TH表达无明显变化(P0.05)。⑧对AchE的影响：与正常组、假手术组比较,模型组大鼠心梗周围区AchE表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组大鼠心梗周围区AchE表达未见明显降低(P0.05)；连夏配方颗粒189.00、94.50 mg/kg组大鼠心梗周围区AchE表达均明显降低(P0.05)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区AchE表达明显降低(P0.05)。⑨对NGF的影响：与正常组、假手术组比较,模型组大鼠心梗周围区NGF表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00 mg/kg组大鼠心梗周围区NGF表达均明显降低(P0.05)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区NGF表达未见明显变化(P0.05)。⑩对TrKA的影响：经比较各组大鼠心肌组织TrKA表达水平的均值发现,模型组大鼠心梗周围区TrKA的表达要高于其余各组。治疗后,各治疗组大鼠心梗周围区TrKA的表达均低于心梗模型组。11对p-TrKA的影响：与正常组、假手术组比较,模型组大鼠心梗周围区p-TrKA表达均显著升高(P0.01)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒189.00、94.50 mg/kg组大鼠心梗周围区p-TrKA表达均显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒189.00 mg/kg组大鼠心梗周围区p-TrKA显著降低(P0.01)。⑩对SH2B的影响：与假手术组比较,模型组大鼠心梗周围区SH2B表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒94.50 mg/kg组大鼠心梗周围区SH2B表达均明显降低(P0.05)。与美托洛尔组比较,连夏配方颗粒94.50 mg/kg组大鼠心梗周围区SH2B均未见明显变化(P0.05)。⑩对Erk1/2的影响：与正常组、假手术组比较,模型组大鼠心梗周围区Erk1/2表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组、连夏配方颗粒94.50、47.25 mg/kg组大鼠心梗周围区Erk1/2表达均明显降低(P0.05)。与美托洛尔组比较,连夏配方颗粒94.50、47.25 mg/kg组大鼠心梗周围区Erk1/2表达均未见明显变化(P0.05)。14对p-Erk1/2的影响：与正常组、假手术组比较,模型组大鼠心梗周围区p-Erkl/2表达明显升高(P0.05)。治疗后,与模型组比较,美托洛尔组大鼠心梗周围区p-Erkl/2表达明显降低(P0.05)；连夏配方颗粒47.25 mg/kg组大鼠心梗周围区p-Erkl/2表达显著降低(P0.01)。与美托洛尔组比较,连夏配方颗粒47.25 mg/kg组大鼠心梗周围区p-Erkl/2表达均未见明显变化(P0.05)结论：①冠脉结扎法心肌梗死模型大鼠心梗周边区TH、AchE表达升高,出现了心脏交感神经和迷走神经的新生和再分布,是研究心梗后心梗周边区心脏自主神经重构的理想动物模型。②美托洛尔可以抑制心梗后心梗周边区交感神经重构,但对心梗后心梗周边区迷走神经重构的作用效果不明显；连夏配方颗粒既可以抑制心梗后心梗周边区交感神经重构,又可以抑制心梗后心梗周边区迷走神经重构。③心梗后大鼠心梗周边区NGF水平升高、TrKA表达增加,SH2B表达上调,进而上调Erkl/2表达,引起TH、AchE合成增加,促进了交感神经和迷走神经的新生和再分布,渐至自主神经重构。这可能是心梗后心梗周边区心脏自主神经重构发生机制之一。④连夏配方颗粒可以通过降低心梗后心梗周边区NGF水平,降低TrKA、SH2B表达,进而下调Erkl/2表达,即下调NGF介导的TrKA信号通路的关键信号靶点的表达,限制TH、AchE合成,减弱心梗周边区交感神经和迷走神经的新生和再分布,抑制心脏自主神经重构。
[Abstract]:After the onset of myocardial infarction, the sympathetic and vagus nerve in the surrounding area of the myocardial infarction will have neural remodeling, which leads to electrocardio physiology and the discrete seriation of repolarization. Secondary disorder becomes the key factor for the occurrence of fatal arrhythmia after myocardial infarction, and the surrounding area of myocardial infarction is the myocardial group after the onset of myocardial infarction. It is imminent to find an effective way to solve the cardiac autonomic nerve reconstruction after myocardial infarction. This paper will discuss three aspects of the cardiac autonomic nerve in the peripheral area of myocardial infarction after myocardial infarction, from the literature review, systematic evaluation and experimental study. The role and mechanism of Lian Xia formula granules in improving cardiac autonomic nerve reconstruction in the peripheral region of myocardial infarction after myocardial infarction were explored in order to find out the relationship between cardiac sympathetic remodeling and arrhythmia after myocardial infarction, and the correlation between nerve growth factor and cardiovascular disease in recent years. The research progress of the physiological function and signal transduction of the nerve growth factor receptor is reviewed in detail. The latest theoretical views, work progress and future research trends in the study of cardiac autonomic nerve reconstruction after myocardial infarction are introduced, and the current research is still discussed. The second part of the system evaluation is to evaluate the efficacy and safety of Huanglian Wendan Decoction in the treatment of arrhythmia. Methods a randomized controlled trial (randomized controlled trial, RCT) of Huanglian Wendan Decoction (Huanglian Wendan Decoction) was collected for the treatment of arrhythmia. According to the standard strict screening, STATA11.0 analysis software was used to carry out Meta meta-analysis. Results Huanglian Wendan Decoction group can obviously improve the outcome indexes of arrhythmia, and its curative effect is higher than that of the control group. The total effective rate of combined effect is [RR=1.267,95%CI:1.157-1.386, P=0.000], the combined effect of comprehensive effect is [RR= 1.340,95%CI:1.111-1.616, P=0.002], and the clinical symptoms are all effective. The rate of rate of combined effect was [RR=1.238,95%CI:1.015-1.510, P=0.035], the early total effective rate of combined effect was [RR=1.256,95%CI:1.003-1.573 in dynamic electrocardiogram room. Conclusion Huanglian Wendan Decoction can obviously improve the total effective efficiency and effective efficiency of the comprehensive curative effect of arrhythmia, the total effective rate of clinical symptoms and the early total effective rate of the dynamic electrocardiogram room. The results and conclusions of this system evaluation still need high quality, and the results and conclusions of this system still need high quality, and the multi center randomized double blind clinical trial to further confirm. The third part of the experimental study aims to observe TH, AchE, NGF, TrKA, SH2B, The protein expression of Erkl/2 mRNA and TH, AchE, NGF, TrKA, p-TrkA, SH2B, Erkl/2, p-Erkl/2 changes in the intervention of cardiac autonomic nerve reconstruction in the periphery of myocardial infarction after myocardial infarction by Lian Xia formula granules. Methods 105 healthy male clean grade 42-49 day old rats were randomly divided into normal group and sham operation according to random number table method. Group, myocardial infarction model group (model group), metoprolol group (Western medicine group), Lian Xia formula granule large, medium and small dose (189 mg/kg, 94.50 mg/kg, 47.25 mg/kg) group total 7 groups, each group of 15 rats. Normal group rats do not do treatment, metoprolol group, large, medium, small dose group of rats were ligated to the left anterior descending coronary artery to establish acute myocardial infarction Rats in the sham operation group were only dressed in the sham operation group without ligature. The normal group, the normal group, the sham operation and the model group were given the normal saline, and the other groups were given the corresponding experimental drugs. After 30 days of continuous perfusion, the rats were anesthetized, and the full amount of myocardium around the same area around the infarct area of the rat was quickly obtained on the ice. The expression level of TH, AchE, NGF, TrKA, SH2B, Erkl/2 in myocardial tissue was detected by Real Time PCR method. The expression of TH mRNA in myocardial tissue was significantly increased (P0.05). After treatment, the expression of TH mRNA in the peripheral zone of myocardial infarction in the group of metoprolol and the group 189.00,94.50,47.25mg/kg of Lian Xia formula was significantly decreased (P0.05). Compared with the metoprolol group, the expression of THmRNA in the area around myocardial infarction in the 189 mg/kg group of Lian Xia formula granule was significantly reduced (P0.0). 5). The effect on AchE mRNA: compared with the normal group and the sham operation group, the expression of AchE mRNA in the myocardial infarction group was significantly increased (P0.01). Compared with the model group, the expression of AchE mRNA in the peripheral region of the metoprolol group was not significantly reduced (P0.05), and even the summer formula 189.00,94.50,47.25 mg/kg group rat myocardial infarction. The expression of AchE mRNA in the surrounding area was significantly decreased (P0.01). Compared with the metoprolol group, the expression of AchE mRNA in the area around myocardial infarction in the 189 mg/kg group was significantly lower (P0.01), and the expression of AchE mRNA expression in the area around myocardial infarction in the 94.50 mg/kg group of Lian Xia formula granule significantly decreased (P0.05). Compared with the myocardial infarction model group, the expression of NGF mRNA in the surrounding area of myocardial infarction was significantly increased (P0.01). After treatment, the expression of NGFmRNA in the group of metoprolol group and the group 189.00,94.50,47.25 mg/kg of Lian Xia formula group was significantly decreased (P0.01). Compared with the metoprolol group, the 189 mg/kg group of Lian Xia formula granule was the group of myocardial infarction week. The expression of NGF mRNA in the peri area was significantly reduced (P0.05). (4) the effect on TrKA mRNA: compared with the normal group and sham operation group, the expression of TrKA mRNA in the myocardial infarction group was significantly increased (P0.01). After treatment, the expression of TrKA mRNA in the metoprolol group and the group 189.00,94.50mg/kg group of Lian Xia formula granules was significant. Lower (P0.01). Compared with the metoprolol group, there was no significant change in the expression of TrKA mRNA in the zone of myocardial infarction in the group 189.00,94.50 mg/kg group of Lian Xia formula group (P0.05). 5. The effect on SH2B mRNA: compared with the normal group and the sham group, the expression of SH2B mRNA expression in the myocardial infarction group was significantly higher (P0.01). After treatment, the ratio of SH2B mRNA to the model group was compared with the model group. Compared with metoprolol group, the expression of SH2B mRNA in the area around myocardial infarction in group 189.00,94.50,47.25mg/kg rats was significantly decreased (P0.01). Compared with metoprolol group, the expression of SH2B mRNA in the area around myocardial infarction of Lian Xia formula 189 mg/kg group was significantly lower (P0.05). 6. The effect of 6 on Erk1/2 mRNA was compared with the normal group and the sham operation group. The expression of Erk1/2 mRNA in the myocardial infarction area of myocardial infarction model group increased significantly (P0.01). After treatment, the expression of Erkl/2 mRNA in the peripheral region of the myocardial infarction group was significantly decreased (P0.01) in the metoprolol group and in the group 189.00,94.50mg/kg of Lian Xia formula granule group (P0.01). The expression of 1/2 mRNA was significantly decreased (P0.01). The effect of TH on TH in the model group was significantly higher than that in the normal group and the sham operation group (P0.05). After treatment, the expression of TH in the peripheral region of the metoprolol group was significantly reduced (P0.05), and the TH table in the 189 mg/kg group of rat myocardial infarction around the summer formula granule group was compared with the model group. Compared with metoprolol group, there was no obvious change in TH expression in the surrounding area of myocardial infarction in group 189 mg/kg of Lian Xia formula group (P0.05). The effect of TH on AchE: compared with the normal group and sham operation group, the expression of AchE in the peripheral region of the model group was significantly increased (P0.05). After treatment, the rats of the model group, the rats of the metoprolol group, were compared with the model group. The expression of AchE in the surrounding area of myocardial infarction was not significantly decreased (P0.05), and the expression of AchE in the area around myocardial infarction in group 189.00,94.50 mg/kg was significantly lower (P0.05). Compared with the metoprolol group, the expression of AchE in the 189 mg/kg group of Lian Xia formula granule group was significantly lower (P0.05). The effect on NGF was in the normal group and the sham operation. Compared with the model group, the expression of NGF in the surrounding area of myocardial infarction was significantly increased (P0.05). Compared with the model group, the expression of NGF in the group of 189 mg/kg rats was significantly lower than that in the model group (P0.05). Compared with the metoprolol group, the expression of NGF in the 189 mg/kg group of Lian Xia formula granule group was not clear. The effect of explicit change (P0.05) on TrKA: the expression of TrKA expression in myocardial tissue of rats in each group was compared. The expression of TrKA in the surrounding area of myocardial infarction in the model group was higher than that of the rest. The expression of TrKA in the peripheral area around the myocardial infarction group was lower than the effect of.11 on p-TrKA in the myocardial infarction group: with the normal group and the sham operation group. Compared with the model group, the expression of p-TrKA in the surrounding area of myocardial infarction was significantly increased (P0.01). After treatment, the expression of p-TrKA in the group of 189.00,94.50 mg/kg rats was significantly decreased (P0.01) in the metoprolol group and in the group 189.00,94.50 mg/kg of Lian Xia formula. Compared with the metoprolol group, Lian Xia formula granule group of 189 mg/kg group of rat heart area p-TrKA The effect on SH2B was significantly reduced (P0.01). Compared with the sham group, the expression of SH2B in the surrounding area of the model group was significantly increased (P0.05). After treatment, the expression of SH2B in the group of metoprolol and the group 94.50 mg/kg of Lian Xia formula group was significantly decreased (P0.05). Compared with the metoprolol group, even the summer formula granule was 9. In 4.50 mg/kg group, there was no significant change in SH2B in the peripheral region of myocardial infarction (P0.05). Compared with the normal group and the sham group, the Erk1/2 expression in the peripheral region of the model group was significantly increased (P0.05). After treatment, the model group, the metoprolol group, the 94.50,47.25 mg/kg group in the group 94.50,47.25 mg/kg group, the Erk1/2 table in the surrounding area of the myocardial infarction group. Compared with the metoprolol group, there was no significant change in the expression of Erk1/2 in the peripheral zone of myocardial infarction (P0.05).14 in the group 94.50,47.25 mg/kg group of Lian Xia formula group (P0.05). Compared with the normal group and the sham group, the expression of p-Erkl/2 in the model group was significantly higher than that in the sham group (P0.05). After treatment, the ratio of the p-Erkl/2 to the model group was compared with the model group. Compared with the metoprolol group, the expression of p-Erkl/2 in the surrounding area of myocardial infarction decreased significantly (P0.05), and the expression of p-Erkl/2 in the area around myocardial infarction in the 47.25 mg/kg group was significantly lower (P0.01). Compared with the metoprolol group, there was no significant change in the expression of p-Erkl/2 in the surrounding area of the 47.25 mg/kg group of Lian Xia formula granule (P0.05): (P0.05) conclusion: 1. The expression of TH and AchE in the peripheral area of myocardial infarction in the rat model of myocardial infarction was elevated, and the new and redistribution of the sympathetic and vagus nerves appeared. It was an ideal animal model to study the reconstruction of autonomic nerve in the periphery of myocardial infarction after myocardial infarction. The effect of vagus nerve reconstruction in the peripheral region of myocardial infarction was not obvious. Even the summer formula could inhibit the sympathetic remodeling in the peripheral region of myocardial infarction and the reconstruction of vagus nerve in the peripheral region of myocardial infarction after myocardial infarction. (3) the level of NGF in the peripheral area of myocardial infarction in the rats after myocardial infarction increased, the expression of TrKA was increased, the expression of SH2B was up-regulated, and then the Erkl/2 table was up-regulated. TH, AchE synthesis increase, promote the regeneration and redistribution of sympathetic and vagus nerve, gradually to autonomic nerve reconstruction. This may be one of the mechanisms of cardiac autonomic remodeling in the peripheral area of myocardial infarction after myocardial infarction. (4) Lian Xia formula granule can reduce the level of NGF in the peripheral region of myocardial infarction after myocardial infarction, reduce TrKA, SH2B expression and down regulation. Erkl/2 expression, that is, downregulates the expression of key signal targets of the TrKA signaling pathway mediated by NGF, restraining TH, AchE synthesis, reducing the regeneration and redistribution of sympathetic and vagus nerve in the peripheral region of the myocardial infarction, and inhibiting the reconstruction of cardiac autonomic nerve.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R285.5

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