天王复心方抗心肌缺血作用及对TLR4-NF κ BP65表达影响的研究

发布时间：2018-11-28 14:03

【摘要】：目的：探讨天王复心方抗心肌缺血作用及其可能作用机理。方法： 1.采用结扎大鼠冠状动脉法复制心肌缺血模型,连续给药21天后,观察各组动物心电图ST段的变化,心肌梗死面积、血清磷酸肌酸激酶(CK)、乳酸脱氢酶(LDH)含量及心肌组织匀浆液SOD、MDA含量。 2.采用异丙肾上腺素多次给药法复制心肌缺血模型,观察天王复心方对心肌缺血模型大鼠心肌损伤程度和损伤范围的影响及血清肌酸磷酸激酶(CK)和乳酸脱氢酶(LDH)变化的影响。 3.以麻醉犬为研究对象,通过八道生理记录仪,观察了天王复心方对动物血压、心率、左室内压、左室内压变化速率、主动脉血流量、冠状动脉血流量、总外周阻力、心肌耗氧量等指标的影响。 4.采用注射肾上腺素与冰水刺激相结合的方法复制大鼠血瘀证模型,观察天王复心方对血瘀证大鼠血液流变学的影响。 5.采用电刺激大鼠颈总动脉血栓形成法,观察不同剂量受试药物对动脉内血栓形成(堵塞动脉)时间(OT)的影响。 6.采用正常大鼠富含血小板的血浆,以5-腺苷二磷酸二钠盐(ADP)为促凝剂,通过血小板聚集仪测血小板聚集曲线,以血小板凝集1分钟(PAGl)、5分钟(PAG5)和最大峰值(PAGM)为观测指标,比较给药各组与空白对照组(生理盐水)血小板聚集率。 7.用结扎大鼠冠状动脉法复制心肌缺血模型,连续给药21天后,采用免疫组化和PCR法观察TLR4-NFκB P65表达的影响结果： 1.结果表明,天王复心方可明显减轻心肌损伤的程度,缩小坏死区范围,降低血浆磷酸肌酸激酶(CK)、乳酸脱氢酶(LDH)含量,提高心肌组织SOD活性,降低MDA含量。提示天王复心方对动物心肌缺血所致的心肌损伤有保护作用。 2.结果表明,天王复心方对异丙肾腺素亚急性心肌缺血动物的心肌损伤有明显的保护作用,可不同程度的的减轻心肌损伤程度,对心肌血清肌酸磷酸激素酶(CK)和乳酸脱氢酶(LDH)水平升高有一定的抑制作用。 3.结果表明,天王复心方高剂量和中剂量可轻度降低血压,增加冠状动脉血流量和主动脉流量,降低总外周阻力,降低心肌耗氧量,对心率没有明显的影响。 4.天王复心方对瘀血证大鼠不同切变率下的全血粘度升高有明显的改善作用,对血浆粘度升高有一定改善作用。 5.天王复心方(2g·kg-1体重和1g·kg-1体重)给药可明显延长电刺激大鼠颈总动脉血栓形成时间(堵塞动脉),表明天王复心方具有抑制血栓形成作用。 6.天王复心方体外100mg/ml和50mg/ml在1min、5min测试点上血小板聚集率明显小于对照组,血小板聚集最大峰值明显低于对照组,提示天王复心方体外给药对ADP所致的血小板聚集均有明显的抑制作用。 7.天王复心方对心肌缺血大鼠炎症调控信号TLR4蛋白与基因、NFκB蛋白表达有抑制作用。结论：天王复心方具有抗凝、抗血栓形成和抗心肌缺血作用,其机制可能与调节炎症信号TLR4-NFκB有关。
[Abstract]:Objective: to investigate the anti-myocardial ischemia effect of Tianwang Fuxin decoction (FXF) and its possible mechanism. Methods: 1. Myocardial ischemia model was established by ligation of coronary artery in rats. After 21 days of continuous administration, the changes of ST segment, myocardial infarction area and serum creatine phosphokinase (CK), were observed in each group. (LDH) content of lactate dehydrogenase and SOD,MDA content of myocardial homogenate. 2. Myocardial ischemia model was established by multiple administration of isoproterenol. To observe the effect of Tianwang Fuxin recipe on myocardial injury degree and damage range and the changes of serum creatine phosphokinase (CK) and lactate dehydrogenase (LDH) (LDH) in rats with myocardial ischemia. 3. The changes of blood pressure, heart rate, left ventricular pressure, aortic blood flow, coronary artery blood flow and total peripheral resistance were observed in anesthetized dogs. The influence of myocardial oxygen consumption and other indexes. 4. The effect of Tianwang Fuxin recipe on hemorheology of rats with blood stasis syndrome was observed by the method of injection of epinephrine and ice water stimulation. 5. The effects of different doses of drugs on the time of thrombosis (clogging) time (OT) of common carotid artery in rats were studied by electrical stimulation. 6. Using platelet-rich plasma of normal rats and 5-adenosine diphosphate disodium salt (ADP) as coagulant, platelet aggregation curve was measured by platelet aggregator, and platelet agglutination was used for 1 minute (PAGl),. Five minutes (PAG5) and maximum peak (PAGM) were measured to compare the platelet aggregation rate between the administration group and the blank control group (normal saline). 7. Myocardial ischemia model was established by ligation of coronary artery in rats. After 21 days of continuous administration, the expression of TLR4-NF 魏 B p65 was observed by immunohistochemical and PCR methods: 1. The results showed that Tianwang Fuxin decoction could obviously reduce the degree of myocardial injury, reduce the area of necrosis, decrease the content of plasma creatine kinase (CK), lactate dehydrogenase (LDH), increase the activity of SOD and decrease the content of MDA. It suggests that Tianwang Fuxin recipe has protective effect on myocardial injury caused by myocardial ischemia. 2. The results showed that Tianwangfuxin recipe had obvious protective effect on myocardial injury of isopropanol subacute myocardial ischemia animals, and could alleviate myocardial injury degree in varying degrees. It can inhibit the increase of serum creatine phosphatase (CK) and lactate dehydrogenase (LDH) levels. 3. The results showed that the high and middle dose of Tianwang Fuxin recipe could decrease blood pressure, increase coronary blood flow and aortic flow, decrease total peripheral resistance and decrease myocardial oxygen consumption, but had no obvious effect on heart rate. 4. Tianwang Fuxin prescription can obviously improve the whole blood viscosity under different shear rates of blood stasis syndrome rats, and improve the plasma viscosity to some extent. 5. Tianwang Fuxin prescription (2 g kg-1 weight and 1 g kg-1 body weight) could significantly prolong the time of thrombosis formation (clogging artery) of rat common carotid artery stimulated by electrical stimulation, which indicated that Tianwang Fuxin prescription could inhibit thrombosis. 6. The platelet aggregation rate of 100mg/ml and 50mg/ml in vitro was significantly lower than that of the control group, and the peak value of platelet aggregation was significantly lower than that of the control group. The results suggest that Tianwang Fuxin prescription has obvious inhibitory effect on platelet aggregation induced by ADP in vitro. 7. Tianwang Fuxin recipe can inhibit the expression of TLR4 protein and gene, NF 魏 B protein of inflammatory regulation signal in myocardial ischemia rats. Conclusion: Tianwang Fuxin recipe has anticoagulant, antithrombotic and anti-myocardial ischemia effects, and its mechanism may be related to the regulation of inflammatory signal TLR4-NF 魏 B.
【学位授予单位】：湖南中医药大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R285

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