5-氨基水杨酸经Nur77调控肺血管内皮细胞重构

发布时间：2018-02-26 19:14

本文关键词： 5-氨基水杨酸 Nur77 内皮细胞重构　出处：《上海交通大学》2015年硕士论文　论文类型：学位论文

【摘要】：第一部分5-氨基水杨酸改善MCT诱导的肺动脉高压大鼠的血流动力学目的:探讨5-氨基水杨酸对野百合碱(MCT)诱导的大鼠肺动脉高压的干预作用。方法:Sprague Dawley(SD)雄性大鼠共60只,随机分为6组,每组10只,分别为:正常对照组(Control组)、肺动脉高压组(PAH组)和5-氨基水杨酸不同剂量治疗组(5ASA-50组,5ASA-100组,5ASA-150组,5ASA-200组)。PAH组、5-氨基水杨酸各剂量治疗组在第1天腹腔注射MCT 50 mg/kg,对照组注射等体积生理盐水。从第2天开始,5-氨基水杨酸各剂量治疗组分别给予5-氨基水杨酸50、100、150 mg/kg/d和200 mg/kg/d灌胃,共30天。第32天,对各组大鼠进行以下检测:(1)统计大鼠的体质量(body mass),计算大鼠体质量增量(weight increment)及其生存率;(2)右心导管法测定肺动脉压力(SPAP,DPAP,MPAP);(3)分离右心室,测定右心室肥厚指数(RVHI)。结果:1.Control组大鼠的体质量和体质量增量均较高于PAH组大鼠(P0.05);5-氨基水杨酸治疗后,除5ASA-150组和5ASA-200组,其余两组大鼠的体质量和体质量增量也较PAH组有明显升高(P0.05);5-ASA治疗组合计生存率较PAH组显著升高(P0.05)。2.PAH组大鼠的SPAP,DPAP,MPAP和RVHI均高于Control组(P0.05);除5ASA-200组的MPAP和RVHI外,其余5-ASA组的SPAP,DPAP,MPAP和RVHI较PAH组有明显降低(P0.05)。结论:5-ASA能够降低野百合碱诱导的肺动脉高压大鼠的肺动脉压力,减轻右心室肥厚,改善生存率。第二部分孤核受体Nur77参与肺血管内皮细胞凋亡和增殖的调控作用目的:探讨Nur77在肺血管内皮细胞凋亡与增殖过程中的调控作用,以及影响Nur77表达的信号通路。方法:通过建立MCT诱导的PAH大鼠模型,采用5-ASA进行药物干预。通过Western Blot、RT-PCR和免疫荧光方法检测孤核受体Nur77在肺动脉内皮细胞内的表达情况。运用HE染色观察肺血管内皮细胞增殖情况。并进一步通过Western Blot和(或)RT-PCR检测炎症介质NF-κBp65、IκBα、COX-2、iNOS,凋亡相关蛋白Bcl-2、caspase-3,增殖细胞核抗原PCNA、MMP-2/-9、TIMP-1/-2以及观察pAkt,pERK,p-p38 MAPK等信号转导因子的表达。结果:1.与PAH组相比,Control组和5-ASA各治疗组大鼠肺血管内皮细胞的Nur77的表达有显著升高(P0.05)。2.Control组和5-ASA各治疗组大鼠的炎症介质NF-κBp65、COX-2、iNOS,抑凋亡蛋白Bcl-2,MMP-2/-9,和信号转导分子pAkt,pERK和p-p38 MAPK的表达有明显下调(P0.05)。3.5-ASA各治疗组的IκBα、caspase-3和TIMP-1/-2的表达较PAH组有明显上调(P0.05)。结论:1.5-氨基水杨酸通过Nur77发挥其抗炎、抑增殖、促凋亡的作用。2.pAkt,pERK和p-p38 MAPK等信号传导通路来调控Nur77的表达。3.Nur77通过调节NF-κB/IκBα来抑制肺血管内皮细胞增殖,促进凋亡。
[Abstract]:Part one: effects of 5-aminosalicylic acid on pulmonary hypertension induced by MCT in rats objective: to investigate the effects of 5-aminosalicylic acid on pulmonary hypertension induced by monocrotaline monocrotaline in 60 male rats. They were randomly divided into 6 groups with 10 rats in each group. Control group, pulmonary hypertension group) and 5-ASA-50 group, 5ASA-100 group, 5ASA-150 group, 5ASA-200 group, PAH group intraperitoneally injected MCT 50 mg / kg on day 1, respectively, and 5-aminosalicylic acid treatment group with different doses of 5-aminosalicylic acid were injected intraperitoneally with MCT 50 mg / kg on day 1. From the second day, each dose of 5-aminosalicylic acid was given intragastric administration of 5-aminosalicylic acid for 50,100,150 mg/kg/d and 200 mg/kg/d, respectively. On the 32nd day (30 days), the body mass of rats was measured as follows: 1) the body mass of rats was calculated, and the survival rate was calculated. (2) right cardiac catheterization was used to measure pulmonary artery pressure (SPAPP, DPAPP, MPAPP) and to separate the right ventricle. Results the body weight and body weight increment of control group were higher than that of PAH group, except 5ASA-150 group and 5ASA-200 group. The body weight and body mass increment of the other two groups were also significantly higher than that of the PAH group. The survival rate of the P0.05-ASA group was significantly higher than that of the PAH group. 2. The SPAPP-DPAPMPAP and RVHI of the rats in the PAH group were significantly higher than those in the Control group, except for MPAP and RVHI in the 5ASA-200 group. Compared with the PAH group, the other 5-ASA groups had significantly lower levels of SPAPPP-DPAPAPP and RVHI than those of the PAH group. Conclusion the ratio of 5 to ASA can reduce the pulmonary artery pressure and the right ventricular hypertrophy in monocrotaline induced pulmonary hypertension rats. To investigate the role of Nur77 in the apoptosis and proliferation of pulmonary vascular endothelial cells. Methods: PAH rat model induced by MCT was established. The expression of solitary nuclear receptor (Nur77) in pulmonary artery endothelial cells was detected by Western blotttin RT-PCR and immunofluorescence. The proliferation of pulmonary vascular endothelial cells was observed by HE staining. Furthermore, the proliferation of pulmonary vascular endothelial cells was observed by Western Blot. RT-PCR was used to detect the expression of signal transduction factors such as NF- 魏 Bp65, I 魏 B 伪, COX-2iNOSs, apoptosis-related protein Bcl-2caspase-3, proliferating cell nuclear antigen PCNAnMMP 2, MMP-2 / 9, TIMP-1 / -2, and the expression of signal transduction factors, such as pAkttpERKCG-p-p38 MAPK. Results 1. Compared with PAH group, the expression of Nur77 in pulmonary vascular endothelial cells of rats in control group and 5-ASA treatment group was observed. The expression of the inflammatory mediators NF- 魏 Bp65-COX-2iNOSand the anti-apoptotic protein Bcl-2MMP 2 / -9, and the signal transduction molecules pAktnpERK and p-p38 MAPK down-regulated the expression of I 魏 B 伪 -caspase-3 and TIMP-1/-2 in the treatment groups of P0.05, 3.5-ASA and 5-ASA were significantly lower than those in the PAH group. Conclusion: 1. 5-aminosalicylic acid can inhibit inflammation through Nur77. Inhibition of proliferation and promotion of apoptosis. 2. Signal transduction pathways such as pAktPERK and p-p38 MAPK regulate the expression of Nur77. 3. Nur77 inhibits the proliferation of pulmonary vascular endothelial cells and promotes apoptosis by regulating NF- 魏 B / I 魏 B 伪.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R544.1

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