漆树酸抑制肿瘤血管新生及早产儿视网膜病变模型的建立

发布时间：2018-11-22 13:36

【摘要】：1.漆树酸通过Src/FAK信号通路抑制肿瘤生长及肿瘤血管新生血管新生是指已经存在于组织的成熟血管内皮细胞通过增殖、迁移以出芽或者嵌入的方式形成新的子代血管的过程。通常情况下只发生在胚胎发育期,生殖周期和伤口愈合期,但在很多病理条件下,如实体肿瘤,糖尿病性视网膜病变,类风湿性关节炎,银屑病和动脉粥样硬化等,疾病与血管新生密切相关。漆树酸,即6-+五烷基水杨酸,提取于银杏和腰果,是组蛋白乙酰转移酶的抑制剂,己被证明具有消炎、抗癌、抗氧化及杀菌等功效。然而,关于漆树酸是否能够调控血管内皮细胞生物学功能以及抑制血管新生的报道鲜见。在本研究中,我们发现漆树酸不仅能在体外细胞水平上抑制血管内皮生长因子(VEGF)诱导的人脐静脉内皮细胞的增殖、迁移、粘附及成管,而且能够在体内水平上显著抑制鸡胚尿囊膜血管形成(n=10)和小鼠角膜血管新生(n=10)。分子机理研究揭示,漆树酸能够浓度梯度依赖性和时间依赖性地抑制Src、FAK激酶的活性,调控Rho-GTPases活性(抑制Rac1-GTPases和Cdc42-GTPases)。每天按2mg/kg的剂量连续皮下给药处理荷瘤裸鼠30d(n=6-7),漆树酸能明显抑制前列腺PC-3肿瘤的体积及重量,但用药处理不影响荷瘤鼠正常体征指标。肿瘤组织免疫组织化学分析发现,给药组肿瘤中CD31和Ki67表达明显降低,提示漆树酸能有效抑制实体肿瘤血管新生和肿瘤细胞增殖。因此,我们的研究结果表明漆树酸能通过抑制Src/FAK信号通路明显抑制肿瘤的生长及肿瘤血管新生。 2.早产儿视网膜病变模型的建立糖尿病性视网膜病变(DR)、老年性黄斑变性(AMD)和早产儿视网膜病变(ROP)这三种常见的眼科疾病严重致盲。致盲的主要原因则是病理性的视网膜血管新生和脉络膜血管新生。为了进一步补充和发展本课题组的血管新生疾病模型,本人参考相关文献,在本实验室条件下,建立了早产儿视网膜病变模型。方法如下,将出生7天后的C57BL/6小鼠用75%的高浓度氧气连续处理5天,再在正常条件下饲养5天并此时行药物处理。将葡聚糖荧光素通过心脏灌流的方式注射到血液中,使周身血管染成绿色。取出眼睛,4%多聚甲醛固定,并在解剖镜下剥离视网膜,荧光显微镜下观察、拍照并统计。结果发现,本方法可成功建立早产儿视网膜病变模型,待测药物藤黄酸和冬凌草甲素等能显著抑制视网膜新生血管的形成。目前该模型已在本研究所内部推广使用。
[Abstract]:1. The inhibition of tumor growth and angiogenesis through Src/FAK signaling pathway refers to the proliferation of mature vascular endothelial cells that already exist in tissues. The process of budding or embedding to form new blood vessels. Usually only during embryonic development, reproductive cycle and wound healing, but in many pathological conditions, such as solid tumors, diabetic retinopathy, rheumatoid arthritis, psoriasis and atherosclerosis, etc. Diseases are closely related to angiogenesis. Lactamic acid, or 6-pentaalkylsalicylic acid, extracted from ginkgo and cashew nuts, is an inhibitor of histone acetyltransferase and has been proven to have anti-inflammatory, anti-cancer, antioxidant and bactericidal effects. However, there are few reports on whether lactoic acid can regulate the biological function of vascular endothelial cells and inhibit angiogenesis. In this study, we found that laccate could not only inhibit the proliferation, migration, adhesion and tube formation of human umbilical vein endothelial cells induced by vascular endothelial growth factor (VEGF) at the cell level in vitro. Moreover, it can significantly inhibit angiogenesis of chick embryo allantoic membrane (nF10) and mouse corneal angiogenesis (NN10) in vivo. The molecular mechanism revealed that lacquer acid could inhibit the activity of Src,FAK kinase in a concentration-gradient and time-dependent manner, and regulate the activity of Rho-GTPases (inhibition of Rac1-GTPases and Cdc42-GTPases). After subcutaneous administration of 2mg/kg every day for 30 days (nb6-7) in nude mice, the volume and weight of prostate PC-3 tumor were significantly inhibited by laccaric acid, but the normal signs of tumor bearing mice were not affected by the treatment. Immunohistochemical analysis of tumor tissue showed that the expression of CD31 and Ki67 was significantly decreased in the treated group, suggesting that lactoxylic acid could effectively inhibit tumor angiogenesis and tumor cell proliferation. Our results suggest that lactoic acid can significantly inhibit tumor growth and tumor angiogenesis by inhibiting Src/FAK signaling pathway. 2. Establishment of retinopathy model in premature infants (DR), age-related macular degeneration (AMD) and premature retinopathy (ROP) are three common ophthalmic diseases which cause severe blindness. The main causes of blindness are pathological retinal angiogenesis and choroidal angiogenesis. In order to further supplement and develop the angiogenic disease model of our group, I have established the model of retinopathy of premature infant under the condition of our laboratory with reference to relevant literature. Methods: C57BL/6 mice were treated with 75% high concentration oxygen for 5 days after birth, then fed under normal conditions for 5 days and then treated with drugs. Dextran fluorescein is injected into the blood through heart perfusion to make the blood vessels green. The eyes were removed, 4% paraformaldehyde was fixed, and the retina was dissected under anatomic microscope, observed under fluorescence microscope, photographed and counted. The results showed that the model of retinopathy of premature infants could be successfully established by this method, and luteinic acid and oridonin could significantly inhibit the formation of retinal neovascularization. At present, the model has been popularized and used in our institute.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R722.6;R-332

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