心肌尔康对L-NAME诱导高血压小鼠的心血管重构的改善作用及其可能机制

发布时间：2018-03-25 11:02

本文选题：心肌尔康　切入点：L-NAME　出处：《安徽医科大学》2017年硕士论文

【摘要】：目的:探讨心肌尔康对L-NAME诱导高血压小鼠心血管重构的影响。探究其可能的机制;比较心肌尔康各剂量组的药理学差异。方法:实验采用60只雄性昆明小鼠,随机分为正常组、模型组、心肌尔康低剂量组、中剂量组、高剂量组和厄贝沙坦组。除正常组外其余5组均以L-NAME诱导建立慢性高血压模型。其中正常组自由饮食饮水,后5组加入L-NAME(2mg/ml),持续给药8wk,自第5wk起每天灌胃给予心肌尔康3.75g/kg,7.5g/kg,15g/kg,厄贝沙坦40mg/kg,每天1次,连续4wk。期间每周测小鼠尾动脉压,实验第8wk末,右颈动脉插管测定血流动力学参数。收集血样品后,充分暴露胸腔,获取心脏和胸主动脉进行相关指标检测。准确称量小鼠体重和心脏质量,计算心脏指数(心脏质量/体质量,HW/BW)。心肌标本进行苏木素-伊红(hematoxylin and eosin,HE)和胶原纤维(Van Gieson,VG)染色,胸主动脉经HE染色,观察其病理学变化。另取一段胸主动脉进行体外内皮依赖性血管舒张功能实验。分别采用硝酸还原酶法、黄嘌呤氧化酶法、硫代巴比妥酸法(thiobarbituric acid,TBA)、测定血清中一氧化氮(nitric oxide,NO)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)含量。双抗体一步夹心法酶联免疫吸附试验(ELISA)检测血清中非对称二甲基精氨酸(asymmetrical dimethylarginine,ADMA)和内皮型NO合酶(endothelial nitric oxide synthase,e NOS)的含量。采用蛋白印迹(western blot,WB)和免疫组化(immunocytochemistry)方法检测心肌组织中e NOS的蛋白表达水平。结果:给药4wk后,与正常组相比,模型组收缩压(systolic blood pressure,SBP)明显升高(P0.01),经心肌尔康各个剂量组以及厄贝沙坦组实验治疗后,心肌尔康中剂量组能够明显抑制SBP、HW/BW、LVSP和+dp/dtmax升高(P0.01,P0.05)。病理学结果显示,经药物实验治疗后,心肌横截面面积(cross-sectional area,CSA)、胶原容积分数(collagen volume fraction,CVF)、血管周胶原面积(perivscular collagen area,PVCA)、中膜厚度(media thickness,MT)均明显减小(P0.01)。表明心肌尔康能够抑制心肌纤维化,改善胸主动脉发生的明显重构。与正常组相比,模型组血清NO和SOD含量明显降低(P0.01),模型组小鼠血清中MDA含量明显升高(P0.01)。与模型组相比,心肌尔康能提升NO含量和SOD活性(P0.01),降低MDA含量(P0.01)和ADMA(P0.05)含量。心肌组织中心肌尔康中剂量组能够明显抑制e NOS蛋白表达降低(P0.05)。结论:心肌尔康能对L-NAME诱导的高血压小鼠发挥保护作用,其可能的药理学机制至少部分与改善内皮细胞功能紊乱,降低ADMA的含量以及降低氧化应激水平相关。
[Abstract]:Objective: to investigate the effect of Xinjierkang on cardiovascular remodeling induced by L-NAME in hypertensive mice, to explore its possible mechanism, and to compare the pharmacological differences of different dose groups of Xinjinerkang. Methods: 60 male Kunming mice were randomly divided into normal group. Model group, low dose group, middle dose group, high dose group and irbesartan group. The model of chronic hypertension was induced by L-NAME in 5 groups except normal group. The latter five groups were treated with L-NAME-2mg / ml of L-NAME-2mg / ml for 8 wks. since 5wk, the rats were given intragastric administration of Myocardial Ercon 3.75g / kg 7.5g / kg 15g / kg, irbesartan 40mg / kg, once a day for 4 wks. during which the caudal arterial pressure of mice was measured every week, and the end of the experiment was the end of 8wk. The hemodynamic parameters were measured by right carotid artery intubation. After collecting blood samples, the thoracic cavity was fully exposed, and the heart and thoracic aorta were obtained to detect the relevant indexes. The weight and heart quality of the mice were weighed accurately. Myocardial samples were stained with hematoxylin and eosin hehe and collagen fiber Van Giesonne VG. The thoracic aorta was stained with HE. Another segment of thoracic aorta was taken for endothelium-dependent vasodilation in vitro. Nitrate reductase method and xanthine oxidase method were used respectively. Thiobarbituric acid (TBAA) was used to determine serum nitric oxide (no), superoxide dismutase (SOD), malondialdehyde (MDA), malondialdehyde (malondialdehyde) (malondialdehyde) (malondialdehyde (MDA)) in serum. Two antibody sandwich enzyme-linked immunosorbent assay (ELISAs) was used to detect asymmetric dimethylarginine (ADMA) and asymmetric dimethylarginine (ADMA) in serum. The expression of e NOS protein in myocardial tissue was detected by Western blotDNA and immunocytochemistryassay methods. Results: after the administration of 4wk, the expression of e NOS in myocardium was detected by Western blotblotBand immunohistochemistry. Results: the expression of e NOS in myocardium was detected by Western blotblotBand immunohistochemical method. Results: after the administration of 4wk, the expression of e NOS in myocardial tissue was detected. Compared with the normal group, systolic blood pressure of SBP in the model group was significantly higher than that in the control group. After experimental treatment with various doses of Myocardial Erkang and irbesartan group, the moderate dose group could significantly inhibit the increase of SBP HW / BWN LVSP and dp/dtmax. The pathological results showed that, After drug treatment, myocardial cross-sectional area, collagen volume fractionation, perivascular collagen area and media thicknessn were significantly decreased (P 0.01). Compared with the normal group, the serum no and SOD levels in the model group were significantly lower than those in the model group, while the MDA content in the serum of the model group was significantly higher than that in the model group, and that in the model group was significantly higher than that in the model group. Xinjierkang can increase the content of no and SOD activity (P0.01), decrease the contents of MDA (P0.01) and ADMA-P0.05). The middle dose group of myocardial tissue can obviously inhibit the expression of e NOS protein and decrease the expression of P0.050.Conclusion: Xenerkang can treat hypertension induced by L-NAME. Mice play a protective role, The pharmacological mechanism may be related at least in part to the improvement of endothelial cell dysfunction, the reduction of ADMA content and the reduction of oxidative stress.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R965

【参考文献】