参草通脉颗粒对心肌梗死后慢性心衰大鼠心肾OPN表达影响的实验研究

发布时间：2018-05-30 23:25

本文选题：参草通脉颗粒 + 慢性心力衰竭　；参考：《辽宁中医药大学》2017年硕士论文

【摘要】：目的:本实验通过建立大鼠心肌梗死后心衰模型,随后分组灌服赖诺普利片和参草通脉颗粒复方的方法,观察骨桥蛋白(osteopontin,OPN)在心肌组织及肾脏组织中的表达,探究慢性心力衰竭(chronic heart failure,CHF)的心肾相关机制,以及参草通脉颗粒对慢性心衰的干预作用,为中医药心肾同治治疗慢性心衰的理念提供科学依据。材料与方法:1.制备慢性心衰大鼠模型:选用SPF级12周龄健康雄性SD大鼠6 0只,体重在230g~250g。其中预留1 5只大鼠进行假手术(开胸但不行冠状动脉结扎),其余4 5只通过结扎大鼠冠状动脉左前降支,配合术后减食、负荷运动的方式制备心肌梗死后慢性心衰大鼠模型。造模后经由多普勒超声心动图计算,若大鼠左室射血分数(EF值)≤5 5%,同时动态心电图显示胸前导联呈ST-T改变,并伴随出现明显精神萎靡,活动度减低,喜欢聚团,毛色无光、脱毛,饮食减少等慢性心衰体征即可纳入实验对象。2.实验分组及喂养:将慢性心衰模型造模成功的2 6只大鼠随机分为模型组(8只)、中药组(9只)、西药组(9只),同时在假手术大鼠中随机选取9只组成假手术组。假手术组及模型组不灌服药物,每天给予常规等容积蒸馏水,剂量为3 ml/次,2次/日;中药组给予参草通脉颗粒灌胃,剂量为每天每公斤9.2mg生药;西药组给予赖诺普利片灌胃,剂量为每天每公斤1.5 mg生药。持续给药喂养28天后再次行彩超检测,检测后将所有大鼠麻醉处死,取其血液、心肌组织、肾脏组织待检。3.检测方法:采用多普勒超声心动图检测并计算各组大鼠LVID(ed)、LVID(es)、EF和FS数值,ELISA法测定大鼠血清中BNP浓度,采用Real-time PCR技术、免疫组化法分别测定假手术组、模型组、中药组和西药组大鼠OPN及OPN mRNA在心肌组织及肾脏组织中的表达水平,并在电镜下观察心肌组织形态结构的改变。结果:1.模型组较假手术组LVID(ed)、LVID(es)均增高(P0.01),EF和FS均减低(P0.01);中药组、西药组与模型组相比LVI D(ed)、LVID(es)减低,EF、FS增高,组间变化显著,差异均具有统计学意义(P0.01);中药组同西药组对比,各指标均无显著差异,无统计学意义,(P0.05)。2.在心肌和肾脏组织中,模型组中OPN mRNA的表达与假手术组相比有着明显升高,P=0.000(P0.01);药物干预的两组较模型组相比OPN mRNA的表达有所下降,差异有统计学意义(P0.01);中药组和西药组中OPN mRNA的表达差异不明显,没有统计学意义,P=0.878(P0.05)。3.较假手术组相对比,模型组中OPN在心肌和肾脏组织中的表达均明显升高,组间差异明显,P=0.000(P0.01);药物治疗的两组较模型组相比,OPN明显降低,差异有统计学意义,(P0.01);将经由药物治疗的中药及西药组进行对比,OPN在心肌和肾脏组织中的表达差异均不明显,无统计学意义,P0.05(心肌组织中p=0.804;肾脏组织中p=0.669)。4.模型组血清中的BNP表达有明显的升高,约是假手术组的2.26倍,与假手术组对照P=0.000,(P0.01),两组间有显著差异;经由药物干预后,BNP的浓度明显降低,与模型组比较,有统计学意义(P0.01);中药组与西药组组间比较效果相仿,差异无统计学意义(P0.05)。5.电镜下观察,假手术组心肌肌原纤维排列致密、整齐,明暗带之间界限清晰整齐,线粒体数量丰富,其基质排列紧密,内脊排列致密整齐,形态清晰可见,糖原颗粒完整丰富;模型组心肌纤维排列紊乱不清,线粒体减少且明显肿胀,内脊结构紊乱或消失,呈明显空炮化,糖原数量也相应减少;中药组和西药组心肌电镜显示,虽然心肌纤维排列紊乱,但勉强可见排列形态,线粒体肿胀,内脊结构紊乱,较模型组中的一系列病理改变有所改善。结论:1.通过结扎大鼠冠状动脉左前降支及术后减食、负荷运动的方法制作心肌梗死后慢性心衰模型方法稳定可行,可应用于慢性心衰的相关研究。2.通过对各组数据进行比较,证明参草通脉颗粒能够通过下调心肌及肾脏组织中的OPN及OPN mRNA的表达,抑制心肌及肾脏的纤维化及重构。3.通过本实验可证明,参草通脉颗粒在慢性心力衰竭治疗的临床应用上是安全有效的,能够达到心肾同治的治疗目的,其治疗效果与赖诺普利相当。
[Abstract]:Objective: to establish a rat model of heart failure after myocardial infarction, and then to group a group of Lisinopril Tablets and Shen Cao Tongmai granule to observe the expression of osteopontin (OPN) in myocardial tissue and renal tissue, and to explore the mechanism of heart and kidney related to chronic heart failure (chronic heart failure, CHF), as well as Shen Cao Tong. The intervention of Mai Granule on chronic heart failure provides a scientific basis for the idea of treating chronic heart failure with Chinese medicine of heart and kidney. Materials and methods: 1. the rat model of chronic heart failure was prepared: 60 healthy male SD rats of SPF grade 12 weeks old were selected, and 15 rats were reserved in 230g~250g. for sham operation (open chest but not coronary artery ligation) The other 45 rats were prepared by ligating the left anterior descending branch of the coronary artery to prepare the rat model of chronic heart failure after myocardial infarction. After the model, the left ventricular ejection fraction (EF) was less than 55%, and the electrocardiogram showed ST-T changes in the prechest lead. The symptoms of chronic heart failure such as aggregation, hair coloring, hair loss, diet reduction and other chronic heart failure were included in.2. experimental group and feeding: 26 rats with chronic heart failure model were randomly divided into model group (8), Chinese medicine group (9) and Western medicine group (9 rats), and 9 randomly selected in the sham operation rats were randomly selected 9. The sham operation group was only made up of the sham operation group. The sham operation group and the model group did not take the medicine. The conventional equal volume distilled water was given daily, the dose was 3 ml/ times, 2 times per day. The Chinese medicine group gave Shencao Tongmai granule gavage, the dose was 9.2mg per kilo a day, and the western medicine group gave Lisinopril Tablets the stomach, the dose was 1.5 mg per kilo a day, and the continuous feeding was 28 days. After the test of color Doppler ultrasound, all rats were killed, and their blood, myocardial tissue and kidney tissue were examined by.3.. The LVID (ED), LVID (ES), EF and FS values were measured and calculated by Doppler echocardiography. ELISA method was used to determine the concentration of BNP in the serum of rats. Real-time PCR technique and immunohistochemical method were used respectively. The expression of OPN and OPN mRNA in the myocardium and kidney tissue of the rats in the sham operation group, the model group, the Chinese medicine group and the western medicine group were measured, and the changes of the morphological structure of the myocardium were observed under the electron microscope. Results: the 1. model group was higher than the sham operation group LVID (ED), LVID (ES) increased (P0.01), EF and FS decreased (P0.01); Chinese medicine group, western medicine group and model group phase Compared with LVI D (ED), LVID (ES), EF, FS increased, the changes between the groups were significant (P0.01), and there was no significant difference between the traditional Chinese medicine group and the western medicine group. (P0.05).2. in the myocardium and kidney tissue, the expression of OPN mRNA in the model group was significantly higher than that in the sham operation group. Compared with the model group, the expression of OPN mRNA decreased in two groups, and the difference was statistically significant (P0.01). There was no significant difference in the expression of OPN mRNA in the traditional Chinese medicine group and the western medicine group. The P=0.878 (P0.05).3. was compared with the sham group. The expression of OPN in the myocardium and kidney tissue in the model group was significantly higher than that in the model group, and the difference between the group and the group was different. Obviously, P=0.000 (P0.01); compared with the model group, the two groups of the drug treatment were significantly lower than the model group, and the difference was statistically significant (P0.01). Compared with the traditional Chinese medicine and the western medicine group, the difference in the expression of OPN in the myocardium and the kidney tissue was not obvious, no systematic significance, P0.05 (p=0.804 in the myocardium in the myocardium, p=0.669 in the kidney).4.. The expression of BNP in the serum of the model group was significantly increased, about 2.26 times of the sham operation group, compared with the sham group P=0.000, (P0.01), and there was a significant difference between the two groups. The concentration of BNP was significantly lower than that in the model group (P0.01), and the effect of the traditional Chinese medicine group and the western medicine group was similar, and the difference was not statistically significant. Under P0.05.5., the myofibrils of the sham operated group were densely arranged and neatly arranged in the sham operation group, with a clear and tidy boundary between the shades, rich mitochondria, tight matrix, dense and neatly arranged internal ridges, a clear and abundant form of the glycogen granules, the disorder of the myofibrils in the model group, and the decrease of mitochondria and obvious swelling of the myocardial fibers in the model group. The internal spinal structure was disorganized or disappeared, and the number of glycogen was reduced. The myocardium of the Chinese medicine group and the western medicine group showed that although the myocardial fibers were arranged in disorder, the arrangement form, the swelling of the mitochondria, the disorder of the internal spinal structure were barely visible, and a series of pathological changes in the model group were improved. Conclusion: 1. by ligating the coronary artery of the rats, the coronary artery was ligated. The method of making the model of chronic heart failure after myocardial infarction is stable and feasible to make the model of chronic heart failure after myocardial infarction. It can be applied to the related study of chronic heart failure. By comparing the data of each group,.2. can inhibit the expression of OPN and OPN mRNA in the myocardium and kidney tissue and inhibit the myocardium and kidney. .3. can be proved to be safe and effective in the clinical application of chronic heart failure in the treatment of chronic heart failure. It can achieve the goal of treating heart and kidney with the same treatment, and the effect of the treatment is equivalent to that of lisinopril.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】