右侧颈交感神经干离断联合不同镇痛药物对大鼠心梗后心室重构的保护作用及机制研究
发布时间：2018-07-24 18:11
【摘要】：第一部分右侧颈交感神经干离断联合不同镇痛药物对大鼠心肌梗死后心室重构的影响目的探索右侧颈交感神经干离断联合不同镇痛药物(帕瑞昔布、氟比洛芬酯、吗啡)对大鼠心肌梗死后心室重构的影响。方法采用结扎心脏冠状动脉前降支的方式构建MI模型及离断右侧颈交感神经干的方法建立TCST模型,36只雄性SD大鼠随机分为6组(n=6):Sham组、MI组、TCST组、TCST+P组、TCST+F组、TCST+M组。MI及右TCST建模后,TCST+P组、TCST+F组、TCST+M组大鼠分别腹腔注射12 mg/kg帕瑞昔布、15 mg/kg氟比洛芬酯、1.2 mg/kg吗啡,2次/d,共3d。于建模处置4周超声心动图检测各组大鼠的心脏功能变化;右侧颈动脉插管的方法测定大鼠血流动力学结果;处死大鼠,取心脏测定左室肥厚指数;HE染色、Masson染色观测梗死周边区心肌组织病理变化和间质胶原蛋白增生情况;Sq RT-PCR法检测梗死周边区心肌ANP m RNA、BNP m RNA含量的变化;q RT-PCR法检测梗死区心肌炎症因子IL-6 m RNA、TNF-αm RNA的表达。结果1心功能结果:与Sham组比较,MI组大鼠心功能指标LVEDd、LVESd显著增加,LVEF、LVFS显著减少(P0.05);与MI组比较,各治疗组LVEDd、LVESd降低,LVEF、LVFS升高(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组LVEDd、LVESd降低,LVEF、LVFS升高,其中TCST+M组心功能改善最显著(P0.05)。2血流动力学结果:与Sham组比较,MI组大鼠心功能指标LVSP、+dp/dtmax显著降低,LVEDP、-dp/dtmax显著升高(P0.05);与MI组比较,各治疗组LVSP、+dp/dtmax升高,LVEDP、-dp/dtmax降低(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组LVSP、+dp/dtmax升高,LVEDP、-dp/dtmax降低,其中TCST+M组血流动力学改善效果最明显(P0.05)。3左室肥厚指数改变:与Sham组比较,MI组体重降低,左室肥厚指数各指标HW/BW、(LV+S)/BW、(LV+S)/HW升高(P0.05);与MI组比较,单用TCST治疗组体重升高,左室肥厚指数各指标降低(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组体重升高,左室肥厚指数各指标降低,其中TCST+M组左室肥厚指数降低最明显(P0.05)。4心肌组织形态学改变:Sham组心肌细胞排列有序,无断裂,显示正常心肌细胞形态;MI组梗死周边区心肌细胞肿胀增粗最显著,排列紊乱,细胞间隙出现大量胶原蛋白增生,呈网状分布,炎性细胞浸润明显,胶原容积分数CVF增多(P0.05);单用TCST治疗组心肌细胞排列紊乱及间质纤维化水平有明显减轻,CVF较MI组降低(P0.05);TCST+P组、TCST+F组、TCST+M组梗死周边区心肌细胞排列较有序,间质纤维降低,炎性细胞浸润进一步减轻,CVF较TCST组降低,其中TCST+M组病理改善效果最明显(P0.05)。5梗死周边区ANP m RNA、BNP m RNA改变:与Sham组比较,MI组梗死周边区心肌肥厚标志物基因ANP m RNA、BNP m RNA的表达升高(P0.05);与MI组比较,TCST干预可降低ANP m RNA、BNP m RNA的表达(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组ANP m RNA、BNP m RNA的表达降低,其中TCST+M组降低最显著(P0.05)。6梗死区IL-6 m RNA、TNF-αm RNA的表达变化:与Sham组比较,MI组梗死区炎症因子IL-6 m RNA、TNF-αm RNA的表达升高(P0.05);与MI组比较,TCST干预可降低IL-6 m RNA、TNF-αm RNA的表达(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组IL-6 m RNA、TNF-αm RNA的表达降低,其中TCST+M组降低最显著(P0.05)。结论1单用TCST及TCST与帕瑞昔布、氟比洛芬酯、吗啡联合治疗均可提高大鼠心肌梗死后心肌舒缩功能,一定程度上减轻心肌肥厚、间质纤维化,延缓心室重构过程。2 TCST与帕瑞昔布、氟比洛芬酯、吗啡联合治疗延缓大鼠心梗后心室重构的效果优于单用TCST治疗,且以TCST联合吗啡改善心室重构的效果最优。3 TCST同帕瑞昔布、氟比洛芬酯、吗啡联合治疗延缓MI后心室重构的机制可能与其强烈的镇痛作用降低了交感神经系统的活性;降低炎症因子IL-6、TNF-α的表达,减少炎症反应有关。第二部分右侧颈交感神经干离断联合不同镇痛药物对大鼠心肌梗死后PI3K/Akt信号通路的影响目的探讨右侧颈交感神经干离断联合不同镇痛药物(帕瑞昔布、氟比洛芬酯、吗啡)对大鼠心肌梗死后PI3K/Akt信号通路的影响。方法采用结扎心脏冠状动脉前降支的方式构建MI模型及离断右侧颈交感神经干的方法建立TCST模型,36只雄性SD大鼠随机分为6组(n=6):Sham组、MI组、TCST组、TCST+P组、TCST+F组、TCST+M组。MI及右TCST建模后,TCST+P组、TCST+F组、TCST+M组大鼠分别腹腔注射12 mg/kg帕瑞昔布、15 mg/kg氟比洛芬酯、1.2 mg/kg吗啡,2次/d,共3d。于建模4周时处死大鼠,取心脏进行Tunel染色,光镜下观测心肌细胞凋亡情况;Western Blot法测定心肌PI3K、Akt、p-Akt、pro-caspase3、caspase3的蛋白表达。结果1心肌细胞凋亡的测定结果:与Sham组比较,MI组在梗死周边区及远隔梗死区组织都存在许多的心肌细胞发生凋亡,周边区凋亡指数AI明显增加(P0.05);与MI组比较,经TCST处理后梗死周边区及远隔梗死区组织心肌细胞凋亡发生明显减少,周边区AI下降(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组梗死周边区及远隔梗死区组织凋亡的心肌细胞进一步降低(P0.05),其中TCST+M组梗死周边区组织AI下降最显著(P0.05)。2 PI3K/Akt信号通路蛋白表达的改变:各组Akt、pro-caspase3蛋白表达无显著差异(P0.05)。与Sham组比较,MI组梗死区组织PI3K、p-Akt蛋白表达均显著下降,caspase3蛋白表达显著上升(P0.05);与MI组比较,TCST干预可提高PI3K、p-Akt蛋白表达,降低caspase3蛋白表达(P0.05);与TCST组比较,TCST+P组、TCST+F组、TCST+M组PI3K、p-Akt蛋白表达均增加,caspase3蛋白表达降低(P0.05)。结论TCST同帕瑞昔布、氟比洛芬酯、吗啡联合治疗延缓MI后心室重构的机制可能通过增强PI3K/Akt抗凋亡信号通路的表达,下调细胞凋亡蛋白caspase3,抑制心肌细胞凋亡有关。
[Abstract]:The effect of the right cervical sympathetic nerve dry dissociation combined with different analgesic drugs on ventricular remodeling after myocardial infarction in rats objective to explore the effect of the right cervical sympathetic trunk disconnection combined with different analgesic drugs (pareoxib, flurbiprofen ester, morphine) on the ventricular restructure after myocardial infarction in rats. Methods the ligature of the coronary artery before the coronary artery was ligated. The MI model and the right cervical sympathetic trunk were constructed by the way of descending branch. 36 male SD rats were randomly divided into 6 groups (n=6): Sham group, MI group, TCST group, TCST+P group, TCST+F group, TCST+M group.MI and right TCST. The rats were intraperitoneally injected with 12 palioxib and 15 flurbiprofen, respectively. Ester, 1.2 mg/kg morphine, 2 times /d, CO 3D. was used to detect cardiac function changes in each group by modeling and disposing 4 weeks echocardiography. The right carotid intubation method was used to determine the hemodynamic results of rats. The rats were killed and the left ventricular hypertrophy index was measured by the heart. HE staining and Masson staining were used to observe the pathological changes of myocardium and interstitial collagen in the surrounding area of the infarct. Sq RT-PCR assay was used to detect the changes of ANP m RNA, BNP m RNA content in the peri infarct zone; Q RT-PCR assay was used to detect the infarct zone myocarditis factor IL-6 m. LVEDd, LVESd, LVEF and LVFS increased (P0.05) in the treatment group. Compared with the TCST group, the TCST+P group, TCST+F group, TCST+M group LVEDd, LVESd decreased, LVEF, and increased. Increase (P0.05); compared with group MI, LVSP, +dp/dtmax, LVEDP, -dp/dtmax decreased (P0.05) in the treatment group, and TCST+P group, TCST+F group and TCST+M group were compared with TCST group. The index of left ventricular hypertrophy index HW/BW, (LV+S) /BW, (LV+S) /HW increased (P0.05). Compared with the MI group, the weight of the left ventricular hypertrophy index was lower (P0.05), and the weight of the TCST+P group, TCST+F group and the left ventricular hypertrophy index were lower than those in the TCST treatment group. .05).4 myocardial histomorphological changes: the myocardial cells in group Sham were arranged in order, without breakage, showing normal cardiac myocyte morphology. The most significant swelling and thickening of myocardial cells in the peripheral area of the MI group was disorderly, a large number of collagen proliferated in the intercellular space, a network distribution, obvious infiltration of inflammatory cells, and increased collagen volume fraction CVF increased (P0.05) alone (P0.05). The myocardial cell arrangement disorder and the level of interstitial fibrosis in the TCST treatment group were significantly reduced, and the CVF was lower than that in the MI group (P0.05). The myocardial cells in the peripheral area of the infarction group, group TCST+P, group TCST+F, and group TCST+M were arranged in a more orderly manner, the interstitial fiber decreased, the infiltration of inflammatory cells was further reduced, and the CVF was lower than the TCST group. The pathological improvement effect of the TCST+M group was most obvious (P0.05).5 ANP m RNA, BNP m RNA changes in the peripheral area of infarct: compared with the Sham group, the expression of ANP m RNA in the peripheral zone of infarct zone in MI group is higher than that in the infarction group. TCST+M group decreased the most significant (P0.05).6 infarct area IL-6 m RNA, TNF- alpha m RNA expression change: compared with the Sham group, the inflammatory factor in the MI group was higher than that in the MI group. The expression of RNA, TNF- alpha m RNA decreased, of which the TCST+M group decreased most significantly (P0.05). Conclusion 1 single use of TCST and TCST with pareoxib, flurbiprofen and morphine can improve myocardial systolic and diastolic function in rats after myocardial infarction, to a certain extent, reduce myocardial hypertrophy, interstitial fibrosis, and delay the process of ventricular remodeling,.2 TCST and pareoxib, fluoro ratio. The effect of combined therapy on ventricular remodeling after myocardial infarction in rats is better than that of TCST alone, and the best effect of TCST combined with morphine to improve ventricular remodeling, the mechanism of.3 TCST with pareoxib, flurbiprofen and morphine for delayed ventricular remodeling after MI may reduce the sympathetic nervous system with its strong analgesic effect. Activity; reducing the expression of inflammatory factors IL-6, TNF- alpha and reducing inflammatory response. Second the effect of the right cervical sympathetic trunk disconnection combined with different analgesic drugs on the PI3K/Akt signaling pathway in rats after myocardial infarction; objective to explore the right cervical sympathetic trunk disconnection combined with different analgesic drugs (pareoxib, flurbiprofen ester, morphine) The effect of PI3K/Akt signaling pathway after myocardial infarction in rats. Methods the MI model and the right cervical sympathetic trunk were established by ligating the anterior descending branch of the heart coronary artery to establish the TCST model. 36 male SD rats were randomly divided into 6 groups (n=6): Sham group, MI group, TCST group, TCST+P group, TCST+F group, TCST+M group.MI and right modeling Group TCST+F, group TCST+F, group TCST+M rats were intraperitoneally injected with 12 mg/kg parinoxib, 15 mg/kg flurbiprofen ester, 1.2 mg/kg morphine and 2 /d. The rats were killed at the 4 week of modeling, and Tunel staining was carried out in the heart. The apoptosis of cardiac myocytes was observed under light microscope. Western Blot method was used to detect the PI3K, Akt, protein expression. The results of apoptosis of 1 cardiac myocytes: compared with the Sham group, there were many cardiomyocytes in the peripheral and distant infarct regions of the MI group, and the apoptosis index AI in the peripheral region was significantly increased (P0.05). Compared with the MI group, the apoptosis of the myocardial cells in the peri infarct zone and the distal septum tissue was significantly decreased after TCST treatment. AI decreased in the border area (P0.05). Compared with the TCST group, the myocardial cells with apoptosis in the peripheral infarct area and the infarct area in group TCST+P, TCST+F and TCST+M were further reduced (P0.05), and the decrease of AI in the tissue around the infarct zone was the most significant (P0.05) the change of the expression of the pathway protein in the.2 PI3K/Akt signal: there was no significant expression of the protein in each group. Difference (P0.05). Compared with group Sham, the expression of PI3K in the infarct area of group MI was significantly decreased, and the expression of Caspase3 protein increased significantly (P0.05). Compared with the MI group, TCST intervention could improve the expression of PI3K, p-Akt protein and decrease the expression of Caspase3 protein. The expression of aspase3 protein was reduced (P0.05). Conclusion the mechanism of TCST with pareoxib, flurbiprofen ester and morphine in the treatment of MI ventricular remodeling may be mediated by enhancing the expression of PI3K/Akt anti apoptotic signaling pathway, lowering the apoptosis protein Caspase3 and inhibiting the apoptosis of cardiac myocytes.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
[Abstract]:The effect of the right cervical sympathetic nerve dry dissociation combined with different analgesic drugs on ventricular remodeling after myocardial infarction in rats objective to explore the effect of the right cervical sympathetic trunk disconnection combined with different analgesic drugs (pareoxib, flurbiprofen ester, morphine) on the ventricular restructure after myocardial infarction in rats. Methods the ligature of the coronary artery before the coronary artery was ligated. The MI model and the right cervical sympathetic trunk were constructed by the way of descending branch. 36 male SD rats were randomly divided into 6 groups (n=6): Sham group, MI group, TCST group, TCST+P group, TCST+F group, TCST+M group.MI and right TCST. The rats were intraperitoneally injected with 12 palioxib and 15 flurbiprofen, respectively. Ester, 1.2 mg/kg morphine, 2 times /d, CO 3D. was used to detect cardiac function changes in each group by modeling and disposing 4 weeks echocardiography. The right carotid intubation method was used to determine the hemodynamic results of rats. The rats were killed and the left ventricular hypertrophy index was measured by the heart. HE staining and Masson staining were used to observe the pathological changes of myocardium and interstitial collagen in the surrounding area of the infarct. Sq RT-PCR assay was used to detect the changes of ANP m RNA, BNP m RNA content in the peri infarct zone; Q RT-PCR assay was used to detect the infarct zone myocarditis factor IL-6 m. LVEDd, LVESd, LVEF and LVFS increased (P0.05) in the treatment group. Compared with the TCST group, the TCST+P group, TCST+F group, TCST+M group LVEDd, LVESd decreased, LVEF, and increased. Increase (P0.05); compared with group MI, LVSP, +dp/dtmax, LVEDP, -dp/dtmax decreased (P0.05) in the treatment group, and TCST+P group, TCST+F group and TCST+M group were compared with TCST group. The index of left ventricular hypertrophy index HW/BW, (LV+S) /BW, (LV+S) /HW increased (P0.05). Compared with the MI group, the weight of the left ventricular hypertrophy index was lower (P0.05), and the weight of the TCST+P group, TCST+F group and the left ventricular hypertrophy index were lower than those in the TCST treatment group. .05).4 myocardial histomorphological changes: the myocardial cells in group Sham were arranged in order, without breakage, showing normal cardiac myocyte morphology. The most significant swelling and thickening of myocardial cells in the peripheral area of the MI group was disorderly, a large number of collagen proliferated in the intercellular space, a network distribution, obvious infiltration of inflammatory cells, and increased collagen volume fraction CVF increased (P0.05) alone (P0.05). The myocardial cell arrangement disorder and the level of interstitial fibrosis in the TCST treatment group were significantly reduced, and the CVF was lower than that in the MI group (P0.05). The myocardial cells in the peripheral area of the infarction group, group TCST+P, group TCST+F, and group TCST+M were arranged in a more orderly manner, the interstitial fiber decreased, the infiltration of inflammatory cells was further reduced, and the CVF was lower than the TCST group. The pathological improvement effect of the TCST+M group was most obvious (P0.05).5 ANP m RNA, BNP m RNA changes in the peripheral area of infarct: compared with the Sham group, the expression of ANP m RNA in the peripheral zone of infarct zone in MI group is higher than that in the infarction group. TCST+M group decreased the most significant (P0.05).6 infarct area IL-6 m RNA, TNF- alpha m RNA expression change: compared with the Sham group, the inflammatory factor in the MI group was higher than that in the MI group. The expression of RNA, TNF- alpha m RNA decreased, of which the TCST+M group decreased most significantly (P0.05). Conclusion 1 single use of TCST and TCST with pareoxib, flurbiprofen and morphine can improve myocardial systolic and diastolic function in rats after myocardial infarction, to a certain extent, reduce myocardial hypertrophy, interstitial fibrosis, and delay the process of ventricular remodeling,.2 TCST and pareoxib, fluoro ratio. The effect of combined therapy on ventricular remodeling after myocardial infarction in rats is better than that of TCST alone, and the best effect of TCST combined with morphine to improve ventricular remodeling, the mechanism of.3 TCST with pareoxib, flurbiprofen and morphine for delayed ventricular remodeling after MI may reduce the sympathetic nervous system with its strong analgesic effect. Activity; reducing the expression of inflammatory factors IL-6, TNF- alpha and reducing inflammatory response. Second the effect of the right cervical sympathetic trunk disconnection combined with different analgesic drugs on the PI3K/Akt signaling pathway in rats after myocardial infarction; objective to explore the right cervical sympathetic trunk disconnection combined with different analgesic drugs (pareoxib, flurbiprofen ester, morphine) The effect of PI3K/Akt signaling pathway after myocardial infarction in rats. Methods the MI model and the right cervical sympathetic trunk were established by ligating the anterior descending branch of the heart coronary artery to establish the TCST model. 36 male SD rats were randomly divided into 6 groups (n=6): Sham group, MI group, TCST group, TCST+P group, TCST+F group, TCST+M group.MI and right modeling Group TCST+F, group TCST+F, group TCST+M rats were intraperitoneally injected with 12 mg/kg parinoxib, 15 mg/kg flurbiprofen ester, 1.2 mg/kg morphine and 2 /d. The rats were killed at the 4 week of modeling, and Tunel staining was carried out in the heart. The apoptosis of cardiac myocytes was observed under light microscope. Western Blot method was used to detect the PI3K, Akt, protein expression. The results of apoptosis of 1 cardiac myocytes: compared with the Sham group, there were many cardiomyocytes in the peripheral and distant infarct regions of the MI group, and the apoptosis index AI in the peripheral region was significantly increased (P0.05). Compared with the MI group, the apoptosis of the myocardial cells in the peri infarct zone and the distal septum tissue was significantly decreased after TCST treatment. AI decreased in the border area (P0.05). Compared with the TCST group, the myocardial cells with apoptosis in the peripheral infarct area and the infarct area in group TCST+P, TCST+F and TCST+M were further reduced (P0.05), and the decrease of AI in the tissue around the infarct zone was the most significant (P0.05) the change of the expression of the pathway protein in the.2 PI3K/Akt signal: there was no significant expression of the protein in each group. Difference (P0.05). Compared with group Sham, the expression of PI3K in the infarct area of group MI was significantly decreased, and the expression of Caspase3 protein increased significantly (P0.05). Compared with the MI group, TCST intervention could improve the expression of PI3K, p-Akt protein and decrease the expression of Caspase3 protein. The expression of aspase3 protein was reduced (P0.05). Conclusion the mechanism of TCST with pareoxib, flurbiprofen ester and morphine in the treatment of MI ventricular remodeling may be mediated by enhancing the expression of PI3K/Akt anti apoptotic signaling pathway, lowering the apoptosis protein Caspase3 and inhibiting the apoptosis of cardiac myocytes.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
【参考文献】
相关期刊论文 前10条
1 雷迁;吕娟娟;梁杰贤;周成斌;王晟;;星状神经节阻滞对兔心肌梗死后心功能的影响[J];岭南心血管病杂志;2016年01期
2 y，
本文编号：2142225
本文链接：https://www.wllwen.com/yixuelunwen/waikelunwen/2142225.html
最近更新
教材专著
