Gas6促进血管新生对缺血性心脏病的保护作用及机制

发布时间：2018-03-18 21:18

本文选题：生长停滞特异性蛋白6(Gas6)　切入点：血管新生　出处：《华中科技大学》2015年博士论文　论文类型：学位论文

【摘要】：背景与目的缺血性心脏病(ischemic heart disease, IHD)是严重威胁老年人群健康的疾病。衰老的机体在心肌缺血情况下,普遍存在血管新生障碍,表现为血管新生能力减弱、密度降低和侧枝循环减少等。心肌血管新生可能是治疗老年患者缺血性心脏病的新途径,但其发生和发展的分子生物学机制尚不明确。有研究表明,生长停滞特异性蛋白6(Gas6)能促进肿瘤血管新生和成熟。我们的研究提示,Gas6是缺血性心脏病的保护因子。该保护作用是否与Gas6促进心肌缺血区血管新生和成熟有关,尚无相关报道。本研究旨在探索以下问题：1.Gas6蛋白是否具有促进血管新生的作用；2.Gas6蛋白促进血管新生的作用机制；3.在人体内抑制血管新生是否导致缺血性心脏病的发生。方法 1.建立来源于人脐带血的内皮祖细胞模型,给予不同浓度的Gas6蛋白干预(25,50,100和200ng/ml)。运用MTT法、Transwell小室迁移和小管形成实验,分别检测细胞增殖、迁移和成管能力。 2.在人内皮祖细胞模型中,给予不同浓度的Gas6蛋白干预(25,50,100和200ng/ml)。并运用P13K抑制剂(LY294002)和ERK的抑制剂(PD98059)进行干预,运用Western blot检测反映EPC增殖功能的MAP和Akt的磷酸化状态；分析对EPC增殖和迁移的影响。 3.在PubMed, EMBASE and Web of Science数据库,检索2012.10月之前发表的所有文献。选择有血管内皮生长因子抗体(贝伐单抗)与对照组治疗肿瘤患者的临床研究进行系统评价,明确血管新生障碍导致患者缺血性心脏病发生的情况。分析来自7个随机对照研究的4,617例患者数据,基于异质性检验的结果运用随机效应模型或固定效应模型,计算所有纳入研究的总缺血性心脏病发生率,相对风险(relative risks, RRs)和95%置信区间(confidence intervals, CIs)。结果 1.Gas6蛋白增强了内皮祖细胞的增殖和迁移能力,并具有剂量依赖性。但是,Gas6蛋白对内皮祖细胞成管未见直接促进作用。 2.Gas6蛋白上调Akt磷酸化的水平,但不上调ERK磷酸化的水平。Gas6蛋白并不促进EPC细胞成管。抑制P13K会显著减少Gas6诱导的内皮祖细胞增殖和迁移,但是抑制ERK并不减少Gas6导致的增殖和迁移。 3.给予血管新生抑制剂治疗的肿瘤患者,缺血性心脏病的发生率是1.0%(95%CI,0.6%-1.4%)。与对照组相比,血管新生抑制剂治疗的肿瘤患者发生缺血性心脏病的风险显著增高(RR=2.49,95%CI：1.37-4.52)。且无论是低剂量(每周2.5mg/kg)还是高剂量(每周5mg/kg)给药,均有缺血性心脏病的风险。(RR分别为2.14,95%CI：1.09-4.19；4.81,95%CI：1.03-22.42)。其中,对于结直肠癌的患者,血管新生抑制剂导致的缺血性心脏病风险比为2.13(95%CI：1.11-4.06)。结论 1.Gas6蛋白增强内皮祖细胞的增殖和迁移功能,但不直接增强成管作用。 2.Gas6蛋白通过激活AKT信号通路增强了内皮祖细胞的增殖和迁移功能。 3.血管新生抑制剂会增加结直肠癌患者缺血性心脏病的发病风险。
[Abstract]:Background and purpose. Ischemic heart disease (IHDD) is a serious threat to the health of the elderly. Myocardial angiogenesis may be a new approach to the treatment of ischemic heart disease in elderly patients, but the molecular biological mechanism of its occurrence and development is unclear. Growth retardation specific protein 6 (Gas6) can promote tumor angiogenesis and maturation. Our study suggests that Gas6 is a protective factor for ischemic heart disease. This protective effect may be related to Gas6 promoting angiogenesis and maturation in ischemic myocardium. The purpose of this study was to investigate whether the Gas6 protein has the role of promoting angiogenesis. 2. The mechanism of the effect of Gas6 protein on angiogenesis. 3. Inhibition of angiogenesis in human body may lead to ischemic heart disease. Method. 1. The endothelial progenitor cell model derived from human umbilical cord blood was established, and different concentrations of Gas6 protein were given to interfere with 2550U / ml and 200ng / ml respectively. The transwell chamber migration and tubule formation were tested by MTT method, and the cell proliferation, migration and tubule formation were measured. 2. In human endothelial progenitor cell model, different concentrations of Gas6 protein were used to interfere with P13K inhibitor LY294002) and ERK inhibitor PD98059). The phosphorylation of MAP and Akt, which reflected the proliferative function of EPC, was detected by Western blot. To analyze the effect on proliferation and migration of EPC. 3. All the literatures published before October 2012 were searched in the PubMed, EMBASE and Web of Science database. The clinical studies of vascular endothelial growth factor antibody (bevacizumab) and control group in the treatment of tumor patients were systematically evaluated. To determine the incidence of ischemic heart disease in patients with angiogenic disorders. Data from 4,617 patients from 7 randomized controlled studies were analyzed. Random effect models or fixed effect models were used based on the results of heterogeneity tests. The incidence of total ischemic heart disease, relative risk relative risks (RRs) and confidence intervals, CIsa, 95% confidence intervals were calculated for all subjects included in the study. Results. 1. Gas6 protein enhanced the proliferation and migration of endothelial progenitor cells in a dose-dependent manner, but Gas6 protein did not directly promote the proliferation and migration of endothelial progenitor cells. 2.Gas6 protein upregulated the level of Akt phosphorylation, but did not up-regulate the level of ERK phosphorylation. Gas6 protein did not promote the proliferation and migration of EPC cells. Inhibition of P13K significantly decreased the proliferation and migration of endothelial progenitor cells induced by Gas6. However, inhibition of ERK does not reduce proliferation and migration induced by Gas6. 3. The incidence of ischemic heart disease in cancer patients treated with angiogenic inhibitors was 1.0% 95% CI 0.6% -1.4% higher than that in the control group. The risk of ischemic heart disease in tumor patients treated with angiogenesis inhibitors was significantly higher than that in cancer patients treated with angiogenic inhibitors. The risk of ischemic heart disease was 2.1495% CI: 1.09-4.19% 95% 95% 95 CI: 1.03-22.42%, regardless of whether it was given at low dose (2.5 mg / kg per week) or high dose (5 mg / kg per week). For patients with colorectal cancer, the risk of ischemic heart disease caused by angiogenesis inhibitors was 2.131.95 CI: 1.11-4.06. Conclusion. 1. Gas6 protein enhanced the proliferation and migration of endothelial progenitor cells, but did not directly enhance the role of tube formation. 2.Gas6 protein enhanced the proliferation and migration of endothelial progenitor cells by activating AKT signaling pathway. 3. Angiogenic inhibitors increase the risk of ischemic heart disease in patients with colorectal cancer.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R541

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