基于《伤寒论》辛开苦降法对心梗后心室重构大鼠的作用及机制研究

发布时间：2018-05-13 21:23

本文选题：心梗后心室重构 + 瘀热证　；参考：《湖北中医药大学》2016年博士论文

【摘要】：理论研究辛开苦降立法于《黄帝内经》,成方完备于《伤寒论》,对后世具有深远的影响。《伤寒论》中辛开苦降法具有调节开阖枢、协调脏腑等功用,广泛运用于寒热错杂痞证、上热下寒证、痰热互结证、瘀热互结证等证候之中,对后世脾胃学说及温病学说的形成有一定的影响。心梗后心室重构是心梗发生后心室出现的一系列变化,应归属于中医学胸痹、心痛、心悸等范畴。中医学对该病的认识,病因主要有外感六淫、情志内伤、饮食不节、年老体衰、久患他病等,病机可归结为本虚标实,本虚是指心、肺、脾、肾等脏气血阴阳的亏损,标实主要是指痰浊、瘀血、水湿的停聚。课题组通过研究历代文献,并结合临床观察,认为随着人们生活方式、饮食、体质等的变化,热邪在该类疾病中的影响越来越突出。因此提出了瘀热相互胶结、相互促进使病情不断演变发展的观点,并总结出以《伤寒论》辛开苦降法为主治疗该类疾病的思路。以辛开苦降法为基础,结合现代的药理研究,拟定了宣痹散结方,其具有活血化瘀、清热散结之功。在多年的临床运用和临床研究中已显示出卓越的疗效。心室重构的现代机制研究主要有血流动力学改变、神经内分泌的激活、炎症反应、细胞凋亡、心肌纤维化等。实验研究目的本课题主要研究基于《伤寒论》辛开苦降法组方的宣痹散结方对心梗后心室重构瘀热证大鼠的防治作用与机制,并为目前正在作为新药开发的该方提供实验研究支持。方法1.宣痹散结方对大鼠疾病和证候的干预研究。对Wistar大鼠采用结扎冠状动脉前降支的方法建立急性心肌梗死模型,存活大鼠喂养4周后即可出现明显的心室重构,对模型B组和宣痹散结方B组两组大鼠再以角叉菜胶和活性干酵母制作瘀热模型。对心室重构的研究主要通过心功能、心脏指数、形态学等参数来观察。对瘀热证的观察主要选用心率、体温、血常规、血清中炎症因子表达水平、血流变等指标。2.宣痹散结方对血清中炎症因子的干预研究。运用ELISA法检测心梗后心室重构模型大鼠血清中IL-6、IL-10、TNF-α等炎症因子的表达水平,以揭示其对大鼠心梗后心室重构的作用机制。3.宣痹散结方对大鼠血清中与纤维化相关因子的干预研究。运用ELISA法检测各组大鼠血清中TGF-β1、MMP-9、TIMP-1、HYP等与纤维化相关因子的表达水平。4.宣痹散结方对心梗后心室重构蛋白表达的干预研究。运用Western blotting法测定各组大鼠心肌组织中COL1、P-smad3、smad7等蛋白的表达。5.宣痹散结方对心梗后心室重构m RNA表达的干预研究。采用实时荧光定量PCR分析法检测各组大鼠心肌组织中NF-κB、smad3、smad7、TGF-β1等因子m RNA的表达。结果对心室重构的影响:1心功能:同模型A组比较,宣痹散结方A组可明显提高大鼠心脏射血分数、但对心室腔无明显干预作用;同模型A组比较,卡托普利可减小舒张期和收缩期的心室腔内径,但对心室射血分数无改善作用。2光镜和电镜显示:模型A组心肌可见明显坏死灶,心肌细胞肿胀,炎性粒细胞浸润,心肌线粒体肿胀。卡托普利和宣痹散结方能够减轻心肌细胞肿胀,减轻炎性细胞浸润,改善心肌线粒体病理变化。3通过Masson染色观察发现卡托普利和宣痹散结方能显著减轻心室重构后心肌纤维化程度。对证候的影响:1心率:同模型A组比较,模型B组心率显著加快,同模型B组比较,宣痹散结方B组心率显著降低。2体温:同模型A组相比,模型B组大鼠在注射活性干酵母后6小时体温明显升高,9小时最明显;同模型B组相比,宣痹散结方B组大鼠体温降低显著。3血常规:同模型A组比较,模型B组大鼠的血红蛋白浓度明显升高,与模型B组相比,宣痹散结方B组大鼠的红细胞压积明显降低。4血流变:同模型A组相比,模型B组大鼠全血黏度中、高切升高显著,同模型B组相比,宣痹散结方B组大鼠的全血黏度低切、中切、红细胞压积明显降低。5血清中炎症因子:同模型A组比较,模型B组大鼠血清中IL-6、TNF-α的表达显著升高,与模型B组相比,宣痹散结方B组的IL-10表达明显升高。作用机制研究:1宣痹散结方对心梗后心室重构大鼠血清中炎症因子IL-6、TNF-α的病理性表达上调具有抑制作用,而对IL-10的表达具有促生作用。2对TGF-β1、MMP-9的表达有抑制作用,而对TIMP-1有促生作用。3对COL1、P-smad3蛋白表达有下调作用,对smad7蛋白表达具有上调作用。4对NF-κBm RNA、TGF-β1m RNA、smad3m RNA的表达有下调作用,对smad7m RNA有上调作用。结论1.以《伤寒论》辛开苦降法为基础的宣痹散结方对心梗后心室重构大鼠的心室重构和瘀热证均能有效地治疗和改善。2.宣痹散结方对NF-k B、IL-6、TNF-α等促使心室重构的炎症因子有下调和抑制作用,而对炎症抑制因子IL-10有促生作用,提示其可通过抑制NF-k B所介导的IL-6等炎症因子所诱发的免疫炎性反应,从而对心室重构和瘀热证起到防治作用。3.宣痹散结方对TGF-β1、MMP-9、COL1、smad3、P-smad3等促使心室重构的纤维化因子有下调和抑制作用,而对抑制纤维化的因子TIMP-1、smad7有促生作用,提示宣痹散结方可作用于TGFβ1-smad通路,从而发挥其抗心室重构的作用。4.宣痹散结方,可作用于NF-k B通路以消除心室重构过程中的免疫炎性反应,并作用于TGFβ1-Smad通路以减轻心肌纤维化,同时对心室重构中的瘀热证候亦具有显著的改善作用,提示心室重构中的NF-k B、TGFβ1-Smad信号转导通路的活化与瘀热证的病机发展具有潜在相关性。
[Abstract]:The theoretical study was made in the "Huangdi Neijing", complete in the "Huangdi Neijing", complete in the "Treatise on typhoid fever", and has a profound influence on later generations. There is a certain influence on the formation of the theory of febrile disease. Ventricular remodeling after myocardial infarction is a series of changes in the ventricular appearance after the onset of myocardial infarction. It should belong to the category of chest pain, heart pain and palpitation in traditional Chinese medicine. The main causes of the disease are six prostitution, internal injury, diet, old age and disease, and the pathogenesis can be attributed to the deficiency of this disease. It refers to the loss of the dirty Qi and blood of the heart, lung, spleen and kidney, which mainly refers to phlegm, blood stasis and water dampness. By studying the literature of the past generation and combining with clinical observation, the subject group thinks that the influence of heat is more and more prominent in this kind of disease with the changes of people's lifestyle, diet and physique. Cementation and mutual promotion of the viewpoint of continuous evolution and development of the disease, and summed up the thought of treating this kind of disease mainly by "the theory of typhoid fever" and "Xin Kai bitterness". Based on the method of "Xin Kai bitterness" and modern pharmacological research, it has drawn up the prescription of Xuan Bi San, which has the function of activating blood and removing stasis and clearing away heat and nodding. It has been used in clinical and clinical studies for many years. The modern mechanism of ventricular remodeling mainly includes hemodynamic changes, neuroendocrine activation, inflammatory response, cell apoptosis, myocardial fibrosis, etc. The effect and mechanism of treatment and research support are provided for the new drug that is currently being developed as a new drug. Method 1. the intervention study on the disease and syndrome of rats in 1. Xuan Bi San. The acute myocardial infarction model was established by ligating the anterior descending coronary artery of the coronary artery in the rats. The survival rats were able to have obvious ventricular remodeling after 4 weeks of feeding. The model B group and the two groups of rats in group B of Xuan Bi Jie Fang were used to make the blood stasis model with carrageenan and active dry yeast. The study of ventricular remodeling was mainly observed by cardiac function, cardiac index, morphology and other parameters. Heart rate, body temperature, blood routine, expression level of inflammatory factors and blood rheology were mainly used to observe the syndrome of blood stasis. The intervention study of the serum inflammatory factors in 2. Xuan Bi Jie Fang. The expression level of IL-6, IL-10, TNF- alpha and other inflammatory factors in the serum of ventricular remodeling model rats after myocardial infarction was detected by ELISA method in order to reveal the mechanism of its effect on ventricular remodeling after myocardial infarction in rats. The intervention study on the serum and fibrosis related factors in the rat serum.3. ELISA method was used to detect the expression level of TGF- beta 1, MMP-9, TIMP-1, HYP and the expression level of fibrosis related factors.4. Xuan Bi Jie Fang on ventricular remodeling after myocardial infarction. The Western blotting method was used to determine the expression of COL1, P-smad3, Smad7 and other proteins in each group of rats. The intervention study of the expression of M RNA in the posterior ventricular remodeling. The expression of NF- kappa B, Smad3, Smad7, TGF- beta 1 and other factors m RNA was detected by real-time fluorescence quantitative PCR analysis. The effect on ventricular remodeling: 1 cardiac function: compared with the model A group, the PBI group could significantly increase the ejection fraction of the rat heart, but to the ventricular cavity No significant intervention; compared with the model A group, Kato Pury could reduce the diastolic and systolic ventricular inner diameter, but there was no improvement on the ventricular ejection fraction by.2 light microscopy and electron microscopy: the myocardium in the model A group showed obvious necrotic foci, myocardial cell swelling, inflammatory granulocyte soaking, myocardial mitochondria swelling. Kato Pury and Xuan Bi Sanjie prescription Can reduce the swelling of myocardial cells, reduce inflammatory cell infiltration, improve the pathological changes of myocardial mitochondria.3 through Masson staining observation found that Kato Pury and Xuan Bi San can significantly reduce the degree of myocardial fibrosis after ventricular remodeling. The impact of syndrome: 1 heart rate: compared with the model A group, the heart rate of the model B group is significantly faster than the model B group, The heart rate of group B of Xuan Bi Sanjie group was significantly reduced by.2 temperature: compared with the model A group, the body temperature of the model B rats increased obviously at 6 hours after the injection of active dry yeast, and the most obvious in the 9 hours. Compared with the model B group, the hypothermia of the rats in group B of the Xuan Bi Sanjie group was significantly.3 blood routine: compared with the model A group, the hemoglobin concentration in the model B group was significantly higher than that of the model A group. Compared with the model B group, the hematocrit of the rats in group B of Xuan Bi Jie Fang group obviously decreased the blood rheology of.4: compared with the model A group, the high shear increased significantly in the whole blood viscosity of the model B group. Compared with the model B group, the whole blood viscosity of the B group of the Xuan Bi Sanjie group was lower than that of the model B group, and the red cell pressure product obviously reduced the inflammatory factors in the.5 serum: the ratio of the model A group to the model A group was significantly lower than that of the model group. The expression of IL-6 and TNF- a in the serum of model B rats increased significantly. Compared with the model B group, the IL-10 expression of B group in the Xuan Bi Sanjie group was significantly increased. The mechanism of action study: 1 Xuan Bi Sanjie prescription inhibited the up regulation of the pathological expression of inflammatory factor IL-6, TNF- a in the serum of ventricular remodeling rats after myocardial infarction, and the expression of IL-10 was promoted. The effect of.2 on the expression of TGF- beta 1 and MMP-9 inhibited the expression of TIMP-1, while.3 had a down-regulation effect on COL1, P-smad3 protein expression and the up regulation of Smad7 protein expression, and the expression of NF- kappa Bm RNA was up. The foundation of Xuan Bi Sanjie Fang can effectively treat and improve the ventricular remodeling and stasis heat syndrome of ventricular remodeling rats after myocardial infarction, which can effectively reduce and inhibit the inflammatory factors of.2. B, IL-6, TNF- A and so on, while promoting the growth of the inflammatory inhibitory factor IL-10, suggesting that it can inhibit IL-6 through the inhibition of NF-k B. The immune inflammatory response induced by inflammatory factors, thus the prevention and control of ventricular remodeling and stasis heat syndrome,.3. Xuan Bi Sanjie Fang has a down-regulation and inhibition effect on TGF- beta 1, MMP-9, COL1, Smad3, P-smad3 and other fibrosis factors that promote ventricular remodeling, but has a stimulating effect on the inhibition of fibrosis, TIMP-1 and Smad7, suggesting that Xuan Bi San can be used as a prescription. The use of TGF beta 1-smad pathway, thus exerting its anti ventricular remodeling effect,.4. Xuan Bi square, can act on the NF-k B pathway to eliminate the inflammatory response in the process of ventricular remodeling, and act on the TGF beta 1-Smad pathway to reduce myocardial fibrosis, and also have a significant improvement in the syndrome of blood stasis in ventricular remodeling, suggesting ventricular weight. The activation of NF-k B, TGF 1-Smad signaling pathway is correlated with pathogenesis of stasis heat syndrome.

【学位授予单位】：湖北中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R222.2

【相似文献】