硼替佐米调控泛素蛋白酶体系统在高肺血流性肺动脉高压中的机制研究

发布时间：2018-02-27 01:23

本文关键词： 肺动脉高压泛素-蛋白酶体系统 Caspase-3 硼替佐米大鼠　出处：《山东大学》2012年硕士论文　论文类型：学位论文

【摘要】：背景与目的肺动脉高压(pulmonary arterial hypertension, PAH)是小儿先天性心脏病的常见并发症之一,以持续增高的肺动脉压并最终导致右心衰为特点的难治性疾病。肺动脉血管内皮损伤、平滑肌细胞过度增殖导致的肺血管重构是肺动脉高压形成的基本病理特征。多数研究认为肺血流增加,血管内皮细胞损伤,平滑肌细胞表型改变、过度增殖、凋亡减少,肺血管床重构和管腔变窄,导致肺动脉高压。肺动脉高压的发病机制涉及多种复杂的细胞因子及其调控网络,其具体发病机制尚不清楚。因此,进一步明确肺动脉高压发病机制,为肺动脉高压提供更有效的治疗方法具有重要意义。本研究的目的是观察在肺动脉高压发生发展过程中泛素蛋白酶体系统的激活情况及硼替佐米对大鼠高血流性肺动脉高压形成的抑制作用,并探讨泛素-蛋白酶体系统及增殖细胞核抗原(PCNA)/Caspase-3在硼替佐米抑制肺动脉高压形成中可能的作用机制。方法45只Wistar大鼠随机分为对照组、分流组、硼替佐米干预组,每组15只。各组于术后第8周测量右心室平均收缩压(RVSP).右心室肥厚指数(RVHI), HE染色观察肺小动脉形态学改变,计算WT%及WA%。采用免疫组化法、Western Blot法检测肺组织中Ub、PCNA、Caspase-3的蛋白表达,VWestern Blot法检测各组大鼠肺组织中NF-κB的活化情况。结果与对照组相比,分流组大鼠肺动脉管壁显著增厚、官腔狭窄,RVSP、RVHI及WT%、WA%增高(P均0.01),Ub、PCNA、Caspase-3蛋白表达增强(P0.05),NF-κB活性增强。与分流组相比,应用硼替佐米干预8周后大鼠肺动脉管壁增厚及管腔狭窄程度明显减轻,(?)RVSP、RVHI及WT%、WA%降低(P0.01),Ub、PCNA蛋白表达减弱(P0.05), Caspase-3蛋白表达增强(P0.05),NF-κB活性受抑制。结论高肺血流性肺动脉高压可激活体内泛素-蛋白酶体系统。硼替佐米可能通过影响NF-κB信号通道来调节肺动脉平滑肌中PCNA/Caspase-3表达而起到抑制高肺血流性肺动脉高压形成的作用。
[Abstract]:Background & objective Pulmonary arterial hypertensionis one of the common complications of congenital heart disease in children. Pulmonary vascular remodeling caused by excessive proliferation of smooth muscle cells is the basic pathological feature of pulmonary hypertension. Pulmonary vascular bed remodeling and narrowing of the lumen lead to pulmonary hypertension. The pathogenesis of pulmonary hypertension involves a variety of complex cytokines and regulatory networks. It is of great significance to clarify the pathogenesis of pulmonary hypertension and to provide more effective treatment for pulmonary hypertension. The aim of this study was to observe the activation of the ubiquitin proteasome system during the development of pulmonary hypertension and the inhibitory effect of bortezomil on the formation of high blood flow pulmonary hypertension in rats. The possible mechanism of ubiquitin proteasome system and proliferating cell nuclear antigen (PCNA) -PCNA / Caspase-3 in the inhibition of pulmonary hypertension by bortezomil was discussed. Methods Forty-five Wistar rats were randomly divided into control group, shunt group and bortezomil intervention group, 15 rats in each group. The mean systolic blood pressure of right ventricle was measured at the 8th week after operation, the right ventricular hypertrophy index was measured and the morphological changes of pulmonary arterioles were observed by HE staining. WT% and WAG were calculated. The expression of Ubhln PCNAfi-Caspase-3 protein in lung tissue was detected by immunohistochemical method and Western Blot method was used to detect the activation of NF- 魏 B in lung tissue of rats in each group. Results compared with the control group, the pulmonary artery wall of shunt group was significantly thickened, and the RVSPN RVHI and WTA increased (P < 0.01) in the shunt group, and the activity of NF- 魏 B was increased in the shunt group compared with the shunt group. The thickness of pulmonary artery wall and the degree of lumen stenosis were significantly reduced after 8 weeks of intervention with bortezomil. The expression of Caspase-3 protein increased and the activity of NF- 魏 B was inhibited. Conclusion High pulmonary flow pulmonary hypertension can activate the ubiquitin proteasome system in vivo. Bortezomil may inhibit the formation of high pulmonary flow pulmonary hypertension by regulating the expression of PCNA/Caspase-3 in pulmonary artery smooth muscle by affecting NF- 魏 B signal channel.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.4

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