黄芩素通过抑制血管重构改善野百合碱诱导的大鼠肺动脉高压

发布时间：2018-05-04 02:07

本文选题：黄芩素 + 肺动脉高压　；参考：《山西医科大学》2017年硕士论文

【摘要】：背景与目的:肺动脉高压(pulmonary artery hypertension,PAH)是一种极具破坏性的心肺疾病,发病率和死亡率均高,其病理特点是肺血管重构造成肺动脉压力(pulmonary arterial pressure,PAP)增高,继而造成右心室肥厚(right ventricular hypertrophy,RVH),最终导致右心室衰竭,甚至死亡。所以,逆转血管重构就是治疗PAH的关键。在肺血管重构过程中,肺动脉不同细胞的增殖/凋亡失衡和炎症反应起着至关重要的作用。黄芩素是一种黄酮类化合物,药理学作用广泛,具有抗增殖,抗炎和心脏保护作用,其作用与MAPK信号通路和NF-κB信号通路紧密相关。但是,黄芩素对PAH是否有治疗作用,其作用机制如何,目前尚不明确。因此本研究旨在探讨黄芩素对MCT诱导的PAH大鼠血管重构的影响及初步机制。方法:1.雄性SD大鼠随机分为4个组,每组10只:对照组、MCT组、MCT+黄芩素(50 mg/kg/d)组、MCT+黄芩素(100 mg/kg/d)组。SD大鼠皮下注射MCT造PAH模型后,于第14天开始灌胃给予不同剂量黄芩素,直至第28天,测定大鼠血流动力学变化后处死大鼠,测定右心室肥厚指数,观察肺形态变化。2.用苏木精-伊红染色法(hematoxylin and eosin high performance liquid,HE)和Masson染色法观察并计算肺小动脉的中膜和肺动脉壁厚度。同时,通过HE染色法测量右心室自由壁厚度和心肌细胞的大小,以观察肺动脉和右心室的重构。3.用HE染色观察肺组织中的炎症反应;用定量反转录聚合酶链式反应(Quantitative reverse transcription-polymerase chain reaction,qRT-PCR)法检测肺组织中肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)、白细胞介素-1β(interleukin-1 beta,il-1β)、白细胞介素-6(interleukin-6,il-6)等炎症因子的mrna表达。4.用tdt介导的缺口末端标记法(tdtmediatednickdutpend-labeling,tunel)观察肺组织细胞凋亡,用westernblot测定肺bcl-2、bax、活化型半胱氨酸蛋白酶-3(cleavedcaspase-3)等凋亡相关蛋白表达。5.用蛋白免疫印迹法测定促丝裂原活化蛋白激酶(mitogen-activatedproteinkinase,mapk)和核因子-κb(nuclearfactor-κb,nf-κb)信号通路的蛋白表达。结果:1.黄芩素可改善mct导致的pah和右心室肥厚。与对照组比较,mct组右心室收缩压显著升高,右心室肥厚指数、右心室/体重的比值均显著增高,右心室自由壁显著增厚,右心室心肌细胞肥大且分布杂乱无序,而黄芩素可有效改善上述病理改变。2.黄芩素可抑制mct诱导的肺动脉重构。he和masson染色法结果显示,mct可导致肺动脉重构和纤维化,肺动脉管壁厚度指数:(肺动脉管壁厚度占管径的百分比)wt%和(管壁面积占血管总面积的百分比)wa%值明显增高,黄芩素可以有效改善上述指标变化。3.黄芩素可抑制mct诱导的炎症反应。he染色结果显示,mct组大鼠肺组织中血管和支气管周围有明显的炎性细胞浸润,而黄芩素减弱此炎症反应。qrt-pcr结果表明,黄芩素可降低mct所致的肺组织炎症细胞因子il-6、tnf-α和il-1βmrna表达的增高。4.黄芩素可抑制mct诱导的细胞凋亡。tunel染色法结果表明,与对照组相比,mct组凋亡细胞平均百分比显著增多,而黄芩素显著减少tunel阳性细胞平均百分比。westernblot结果表明,黄芩素可降低mct所致肺组织bax/bcl-2比率和cleavedcaspase-3表达的上调。5.黄芩素可抑制mct诱导的mapk和nf-κb信号通路的激活。westernblot结果表明,与对照组相比,mct组大鼠的肺组织中p38,erk和jnk磷酸化显著增强,而黄芩素给药组中p38,erk和jnk磷酸化显著被抑制。与对照组相比,mct组大鼠肺组织nf-κbp65表达显著增高,而黄芩素可显著抑制mct诱导的nf-κb上调。结论:1.黄芩素可改善MCT诱导的PAH和右心室肥厚,其保护作用可能与其抑制肺血管重构有关。2.黄芩素抑制MCT诱导的PAH分子机制可能与其抑制MAPK和NF-κB信号通路有关。
[Abstract]:Background and purpose: pulmonary artery hypertension (PAH) is a highly destructive cardiopulmonary disease with high morbidity and mortality. Its pathological feature is that pulmonary vascular remodeling causes increased pulmonary arterial pressure (pulmonary arterial pressure, PAP), and then causes right ventricular hypertrophy (right ventricular hypertrophy, RVH). It leads to right ventricular failure and even death. Therefore, reversing vascular remodeling is the key to the treatment of PAH. In the process of pulmonary vascular remodeling, the proliferation / apoptosis imbalance and inflammatory response of different cells in the pulmonary artery play a vital role. The effect is closely related to the MAPK signaling pathway and the NF- kappa B signaling pathway. However, it is not clear whether baicalein has a therapeutic effect on PAH, and the effect of baicalein on vascular remodeling induced by MCT in PAH rats and its preliminary mechanism. Methods: 1. male SD rats were randomly divided into 4 groups, each group was 10. Only: the control group, MCT group, MCT+ baicalein (50 mg/kg/d) group, MCT+ baicalein (100 mg/kg/d) group.SD rats were subcutaneously injected with MCT to build the PAH model, and the rats were given different doses of baicalein on the fourteenth day, until twenty-eighth days. After the hemodynamic changes in the rats, the rats were killed and the right ventricular hypertrophy index was measured, and the pulmonary morphologic changes were observed with hematoxylin in.2.. The hematoxylin and eosin high performance liquid, HE, and Masson staining were used to observe and calculate the thickness of the middle and pulmonary artery walls of the pulmonary arteriole. Meanwhile, the free wall thickness of the right ventricle and the size of the cardiac myocytes were measured by the HE staining method to observe the reconstructive.3. of the pulmonary and right ventricles by HE staining to observe the inflammation in the lung tissue. A quantitative reverse transcription polymerase chain reaction (Quantitative reverse transcription-polymerase chain reaction, qRT-PCR) was used to detect tumor necrosis factor - alpha (tumor necrosis factor alpha, TNF- alpha), interleukin -1 beta, interleukin, and other inflammatory factors in lung tissue. MRNA expression.4. uses TDT mediated tdtmediatednickdutpend-labeling (TUNEL) to observe the apoptosis of lung tissue cells. The expression of apoptosis related proteins such as Bcl-2, Bax, and activated cysteine protease -3 (cleavedcaspase-3) in lung Bcl-2, Bax, and activated cysteine proteinase -3 (cleavedcaspase-3) is determined by Westernblot. N-activatedproteinkinase, MAPK) and nuclear factor kappa B (nuclearfactor- kappa B, nf- kappa B) protein expression. Results: 1. baicalein can improve MCT induced PAH and right ventricular hypertrophy. Compared with the control group, the right ventricular systolic pressure increased significantly in the MCT group, the right ventricular hypertrophy index, the right ventricular / body weight ratio increased significantly, the right ventricular free wall Significantly thickening, right ventricular myocytes hypertrophy and distribution disorder, and baicalein can effectively improve the above pathological changes,.2. baicalein can inhibit MCT induced pulmonary artery remodeling,.He and Masson staining results show that MCT can lead to pulmonary artery remodeling and fibrosis, pulmonary artery wall thickness index: (pulmonary artery wall thickness accounts for 100% of the diameter of the pipe. The wa% value of wt% and (the percentage of the wall area in total vascular area) increased significantly, and baicalein could effectively improve the changes of the above indexes..3. baicalein could inhibit the inflammatory reaction induced by MCT,.He staining results showed that there was obvious inflammatory cell infiltration around the blood vessels and bronchi in the lung tissue of the MCT group, and baicalein weakened the inflammatory reaction.Q RT-PCR results showed that baicalein could reduce the inflammatory cytokine IL-6 of lung tissue caused by MCT, the increase of tnf- A and IL-1 beta mRNA expression of.4. baicalein inhibited MCT induced apoptosis by.Tunel staining. Compared with the control group, the average percentage of apoptotic cells in the MCT group increased significantly, while baicalein significantly reduced the tunel positive cell level. The results of average percentage of.Westernblot showed that baicalein could reduce the bax/bcl-2 ratio of lung tissue and the up regulation of cleavedcaspase-3 expression by MCT. The activation of MAPK and nf- kappa B signaling pathway induced by MCT indicated that MCT induced MAPK and nf- kappa B signaling pathway activated.Westernblot. The phosphorylation of p38, ERK and JNK in baicalein group was significantly inhibited. Compared with the control group, the expression of nf- kappa bp65 in the lung tissue of MCT rats increased significantly, and baicalein significantly inhibited the MCT induced up regulation of nf- kappa B. Conclusion: 1. baicalein can improve PAH and right ventricular hypertrophy induced by MCT. The protective effect of baicalein may be related to the inhibition of pulmonary vascular remodeling. The molecular mechanism of baicalein inhibiting MCT induced PAH may be related to its inhibition of MAPK and NF- kappa B signaling pathway.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R544.1

【相似文献】