去肾交感神经术对心衰犬心功能及凋亡相关基因表达的影响
发布时间：2018-08-08 12:07
【摘要】：背景心肌细胞凋亡作为心肌梗死后心力衰竭(MI-HF)的主要发病机制之一,在HF的发生发展及进程中至关重要。MI后缺血缺氧、氧化应激、炎症介质的产生及异常的神经体液机制等局部恶劣微环境的作用下,细胞发生凋亡,可导致心肌有效收缩单位丧失、胶原纤维沉积,促进了心室重构及心室功能的恶化进展。心肌细胞的丢失与心功能不可逆性下降相关,因此抑制心梗后心肌凋亡有关的不良刺激因素,进而延缓心脏重构及心室扩张的进程,改善并提高心功能,对于心衰患者的治疗具备临床指导意义。目前,去肾交感神经术(RDN)正成为治疗心衰的热点,已有研究显示RDN可以降低心室起搏所致房颤动物模型的心肌细胞凋亡现象进而改善心功能,但对于RDN是否可以调节MI-HF的细胞凋亡现象及其相关机制尚不明确。目的本实验旨在探讨RDN对MI-HF犬心功能及细胞凋亡相关基因表达的影响。方法将18只健康杂种犬随机分为正常组、模型组及治疗组,其中模型组及治疗组采用无水酒精栓塞法制作心梗后心衰模型。治疗组犬在心衰造模成功后行双侧肾动脉消融治疗,模型组犬行肾动脉造影。测量基线、肾动脉消融前及消融术后4周心功能参数;ELISA法检测血清NT-Pro BNP水平;心肌纤维化水平应用Masson染色法进行观察评估;通过TUNEL法观察心肌细胞凋亡状况并计算凋亡指数(AI),RT-PCR及WB法检测心肌凋亡指标Bcl-2、Bax、Caspase-3及GRP78相对表达量;肾动脉HE染色评估RDN术去神经化的有效性;检测血清肌酐水平,以评估手术操作安全性。结果1、基线时三组实验犬在体重、HR、LVEDD、LVESD、LVEF、LVEDP及LVSP两两比较差异均无统计学意义(P0.05)。肾动脉消融前,与正常犬比较,心衰犬(模型组+治疗组)LVEDD、LVESD及LVEDP增加(P0.05),而LVEF、LVSP下降(P0.05);模型组与治疗组相比,各心功能参数均无统计学差异(P0.05)。消融术后4周,与模型组比较,经RDN治疗后LVEDD、LVESD及LVEDP降低(P0.05),LVEF及LVSP增加(P0.05),而模型组+治疗组犬较正常组犬LVEDD、LVESD及LVEDP增加(P0.05),LVEF降低(P0.05),模型组LVSP较正常组降低(P0.05),而治疗组犬LVSP较术前稍有升高,与正常组无统计学差异(P0.05)。2、基线时三组实验犬血清NT-Pro BNP水平无统计学差异(P0.05)。肾动脉消融前,心衰犬较正常组犬血清NT-Pro BNP值增加(P0.05);模型组与治疗组相比,NT-Pro BNP水平无统计学差异(P0.05)。消融后4周,与模型组比较,经RDN治疗后血清NT-Pro BNP值降低(P0.05);而模型组+治疗组犬较正常组犬NT-Pro BNP水平增加(P0.05)。3、消融后4周,与正常组犬比较,心衰犬心肌组织Bcl-2 m RNA及蛋白含量表达下降(P0.05),Bax、caspase-3、GRP78 m RNA及蛋白表达与心肌凋亡指数升高(P0.05);与模型组犬相比,经RDN治疗后心肌Bcl-2 m RNA及蛋白表达上调(P0.05),Bax、caspase-3、GRP78 m RNA及蛋白表达与凋亡指数下调(P0.05)。4、消融后4周,HE染色显示RDN术后肾动脉血管外膜神经纤维轴突缺失破化。5、基线及肾动脉消融术前,三组实验犬血清肌酐值无统计学差异(P0.05);消融后4周,与正常组比较,模型组犬血清肌酐值升高(P0.05),而治疗组肌酐值变化无统计学差异(P0.05)。结论RDN可改善心肌梗死后心衰犬的心功能,我们推测可能与RDN上调抗凋亡基因Bcl-2的表达,下调抑凋亡基因Bax、Caspase-3及GRP78表达水平,进而降低细胞凋亡程度有关。
[Abstract]:Background cardiac myocyte apoptosis is one of the main pathogenesis of heart failure (MI-HF) after myocardial infarction. In the development and process of HF, the cell apoptosis is induced by ischemia and hypoxia, oxidative stress, the production of inflammatory mediators and abnormal neurohumoral mechanism, which can lead to the effective myocardial harvest of the myocardium. Loss of units and deposition of collagen fibers contribute to the deterioration of ventricular remodeling and ventricular function. The loss of cardiac myocytes is associated with a decline in cardiac function irreversibility, thus inhibiting the adverse stimulus related to myocardial apoptosis after myocardial infarction, further delaying the process of cardiac remodeling and ventricular dilatation, and improving cardiac function in patients with heart failure. At present, RDN is becoming a hot spot in the treatment of heart failure. Research has shown that RDN can reduce cardiac apoptosis in the animal model of atrial fibrillation induced by ventricular pacing and then improve cardiac function, but it is still unclear whether RDN can regulate the apoptosis of MI-HF and its related mechanisms. The purpose of this experiment was to investigate the effect of RDN on the cardiac function and the expression of apoptosis related genes in MI-HF dogs. Methods 18 healthy hybrid dogs were randomly divided into normal group, model group and treatment group. The model group and the treatment group were used to make heart failure model after myocardial infarction. Renal artery ablation was used in the treatment of renal artery. The heart function parameters were measured at baseline, 4 weeks after renal artery ablation and 4 weeks after ablation. The level of serum NT-Pro BNP was measured and the level of myocardial fibrosis was evaluated by Masson staining. The apoptosis of myocardial cells was observed by TUNEL, and the apoptosis index (AI), RT-PCR and WB were calculated. The relative expressions of Bcl-2, Bax, Caspase-3 and GRP78 were detected by the method of myocardial apoptosis, and the renal artery HE staining was used to evaluate the effectiveness of RDN neurosurgery, and the level of serum creatinine was detected to evaluate the safety of operation. Results 1, there were no significant differences in weight, HR, LVEDD, LVESD, LVEF, LVEDP, and LVSP 22 in the three groups of experimental dogs at baseline. Before renal artery ablation, compared with normal dogs, LVEDD, LVESD and LVEDP increased (P0.05) in heart failure dogs (model group + treatment group), and LVEF, LVSP decreased (P0.05). Compared with the treatment group, there was no statistical difference between the model group and the treatment group (P0.05). Compared with the model group, LVEDD, LVESD and LVEDP decreased after 4 weeks of RDN treatment. In model group + treatment group, LVEDD, LVESD and LVEDP increased (P0.05), LVEF decreased (P0.05), LVSP in model group was lower than that of normal group (P0.05), but the LVSP in the treatment group was slightly higher than that before the operation. There was no statistical difference between the treatment group and the normal group (P0.05).2. There was no statistical difference between the three groups of dogs. Before the renal artery ablation, the NT-Pro BNP value of heart failure dog was increased (P0.05) than that of the normal group. Compared with the treatment group, the level of NT-Pro BNP was not statistically significant (P0.05). After 4 weeks of ablation, the serum NT-Pro BNP value of the model group decreased after RDN treatment (P0.05), while the model group + treatment group was more than the normal group. 4 weeks after ablation, the expression of Bcl-2 m RNA and protein content in heart failure dogs decreased (P0.05), Bax, Caspase-3, GRP78 m RNA and protein expression and the increase of myocardial apoptosis index (P0.05). Down regulation of apoptosis index (P0.05).4, 4 weeks after ablation, HE staining showed that the renal artery epicardial nerve fiber axonal deletion of the renal artery was.5 after RDN. Before the baseline and the renal artery ablation, the serum creatinine value of the three experimental dogs was not statistically different (P0.05). After 4 weeks, the serum creatinine value of the model group was elevated (P0.05), and the treatment group was compared with the normal group (P0.05). There is no significant difference in creatinine value (P0.05). Conclusion RDN can improve cardiac function of heart failure dogs after myocardial infarction. We speculate that RDN may up regulate the expression of anti apoptotic gene Bcl-2, down regulation of apoptosis gene Bax, Caspase-3 and GRP78 expression level, and then decrease the degree of apoptosis.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.6
本文编号：2171739
[Abstract]:Background cardiac myocyte apoptosis is one of the main pathogenesis of heart failure (MI-HF) after myocardial infarction. In the development and process of HF, the cell apoptosis is induced by ischemia and hypoxia, oxidative stress, the production of inflammatory mediators and abnormal neurohumoral mechanism, which can lead to the effective myocardial harvest of the myocardium. Loss of units and deposition of collagen fibers contribute to the deterioration of ventricular remodeling and ventricular function. The loss of cardiac myocytes is associated with a decline in cardiac function irreversibility, thus inhibiting the adverse stimulus related to myocardial apoptosis after myocardial infarction, further delaying the process of cardiac remodeling and ventricular dilatation, and improving cardiac function in patients with heart failure. At present, RDN is becoming a hot spot in the treatment of heart failure. Research has shown that RDN can reduce cardiac apoptosis in the animal model of atrial fibrillation induced by ventricular pacing and then improve cardiac function, but it is still unclear whether RDN can regulate the apoptosis of MI-HF and its related mechanisms. The purpose of this experiment was to investigate the effect of RDN on the cardiac function and the expression of apoptosis related genes in MI-HF dogs. Methods 18 healthy hybrid dogs were randomly divided into normal group, model group and treatment group. The model group and the treatment group were used to make heart failure model after myocardial infarction. Renal artery ablation was used in the treatment of renal artery. The heart function parameters were measured at baseline, 4 weeks after renal artery ablation and 4 weeks after ablation. The level of serum NT-Pro BNP was measured and the level of myocardial fibrosis was evaluated by Masson staining. The apoptosis of myocardial cells was observed by TUNEL, and the apoptosis index (AI), RT-PCR and WB were calculated. The relative expressions of Bcl-2, Bax, Caspase-3 and GRP78 were detected by the method of myocardial apoptosis, and the renal artery HE staining was used to evaluate the effectiveness of RDN neurosurgery, and the level of serum creatinine was detected to evaluate the safety of operation. Results 1, there were no significant differences in weight, HR, LVEDD, LVESD, LVEF, LVEDP, and LVSP 22 in the three groups of experimental dogs at baseline. Before renal artery ablation, compared with normal dogs, LVEDD, LVESD and LVEDP increased (P0.05) in heart failure dogs (model group + treatment group), and LVEF, LVSP decreased (P0.05). Compared with the treatment group, there was no statistical difference between the model group and the treatment group (P0.05). Compared with the model group, LVEDD, LVESD and LVEDP decreased after 4 weeks of RDN treatment. In model group + treatment group, LVEDD, LVESD and LVEDP increased (P0.05), LVEF decreased (P0.05), LVSP in model group was lower than that of normal group (P0.05), but the LVSP in the treatment group was slightly higher than that before the operation. There was no statistical difference between the treatment group and the normal group (P0.05).2. There was no statistical difference between the three groups of dogs. Before the renal artery ablation, the NT-Pro BNP value of heart failure dog was increased (P0.05) than that of the normal group. Compared with the treatment group, the level of NT-Pro BNP was not statistically significant (P0.05). After 4 weeks of ablation, the serum NT-Pro BNP value of the model group decreased after RDN treatment (P0.05), while the model group + treatment group was more than the normal group. 4 weeks after ablation, the expression of Bcl-2 m RNA and protein content in heart failure dogs decreased (P0.05), Bax, Caspase-3, GRP78 m RNA and protein expression and the increase of myocardial apoptosis index (P0.05). Down regulation of apoptosis index (P0.05).4, 4 weeks after ablation, HE staining showed that the renal artery epicardial nerve fiber axonal deletion of the renal artery was.5 after RDN. Before the baseline and the renal artery ablation, the serum creatinine value of the three experimental dogs was not statistically different (P0.05). After 4 weeks, the serum creatinine value of the model group was elevated (P0.05), and the treatment group was compared with the normal group (P0.05). There is no significant difference in creatinine value (P0.05). Conclusion RDN can improve cardiac function of heart failure dogs after myocardial infarction. We speculate that RDN may up regulate the expression of anti apoptotic gene Bcl-2, down regulation of apoptosis gene Bax, Caspase-3 and GRP78 expression level, and then decrease the degree of apoptosis.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R541.6
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