球结膜微循环在兔系统性硬化肺动脉高压中的意义及VEGF的相关性研究

发布时间：2018-05-28 23:05

本文选题：系统性硬化 + 肺动脉高压　；参考：《南昌大学》2017年硕士论文

【摘要】：背景和目的:肺动脉高压(pulmonary arterial hypertension,PAH)是一类病因复杂、预后较差的疾病。特别是结缔组织病(connective tissue disease,CTD)相关PAH,其起病隐匿,诊断不充分,在临床诊疗中仍是难点。微循环障碍是系统性硬化(systemic sclerosis,SSc)发病机制中的中心环节,两者间的相关性也越来越被认可。由于临床研究的众多局限,动物模型对疾病发病机制及诊疗手段的探索提供了极大的帮助。本研究拟用博来霉素(bleomycin,BLM)联合野百合碱(monocrotaline,MCT)诱导模拟SSc合并PAH发病机制的兔模型,探讨球结膜微循环与PAH的联系,及其在疾病中的早期预测价值。同时,血管内皮生长因子(vascular endothelial growth factor,VEGF)既往已被多个研究证实在微循环障碍、血管重塑中发挥重要作用。本文试图在动物模型中,对PAH、球结膜微循环分析及VEGF水平的相关性进行讨论,进一步探索球结膜微循环在SSc合并PAH中的意义。方法:1.选取新西兰雄兔24只,随机分为三组(溶剂对照组、模型组、激素组),每组8只。模型组及激素组分别一次性腹腔注射1%的MCT50mg/kg及每日腹部皮下注射BLM 100ul(浓度1mg/ml),激素组同时每日腹腔注射甲泼尼龙(methylprednisolone,MP)2.6mg/kg(浓度1mg/ml),三周后停止给药。随后,利用右心导管测量平均肺动脉压(mean pulmonary arterial pressure,mPAP),与溶剂对照组对比,判断造模是否成功。并通过测量右心室肥大指数(right ventricular hypertrophy index,RVHI)及完善组织病理,进一步评估模型是否符合SSc合并PAH病理特点。2.给药前后进行球结膜微循环分析,利用酶联免疫吸附法(ELISA)检测血清VEGF表达。分析球结膜微循环积分、VEGF浓度及mPAP的相关性,并利用ROC曲线判断球结膜微循环积分对SSc合并PAH的诊断价值。结果:1.模型组mPAP及RVHI均显著高于溶剂对照组和激素组,有统计学意义(P0.05)。2.组织病理:皮肤染色显示,模型组皮损处表皮萎缩并角化过度,真皮层出现少量淋巴细胞、浆细胞浸润;肺组织染色显示,模型组肺泡内可见大量出血,细小动脉肌化并不规则增厚,形成丛样结构,伴有淋巴细胞、浆细胞浸润。3.结膜微循环分析显示,模型组总积分较其他两组有明显差异(P0.05),主要表现在血管增粗、血管扭曲、网状畸形、出血、血管色调等方面,以上项目均与mPAP成正相关(P0.05)。4.球结膜微循环总积分ROC曲线分析,曲线下面积为0.852(P0.05)。总分5.2分对应的正确诊断指数最大,把球结膜微循环总积分≥5.2分作为辅助诊断PAH的分界点,其敏感度为88.9%,特异度为75.0%。5.模型组血清VEGF浓度较其他两组有显著差异(P0.05),并与球结膜微循环总积分及mPAP成正相关(P0.05)。结论:1.新西兰雄兔利用BLM联合MC T注射可以成功模拟SSc合并PAH模型。2.SSc合并PAH存在球结膜微循环障碍,其严重程度与mPAP成正相关,特别是在血管增粗、血管扭曲、出血、网状畸形、血管色调等方面。3.球结膜微循环总积分≥5.2分可以作为辅助诊断PAH的分界点,其敏感度为88.9%,特异度为75.0%。4.SSc合并PAH中血清VEGF水平有所升高,并与球结膜微循环总分、mPAP有一定的相关性。
[Abstract]:Background and purpose: pulmonary arterial hypertension (PAH) is a kind of disease with complex causes and poor prognosis. Especially the connective tissue disease (CTD) related PAH, its onset is hidden, and the diagnosis is not sufficient. It is still difficult in clinical diagnosis and treatment. Microcirculation disorder is systemic sclerosis (systemic sclerosis, SSc). The central link in the pathogenesis is becoming more and more recognized. Because of the numerous limitations of the clinical research, animal models provide great help to the pathogenesis and the exploration of the means of diagnosis and treatment. This study is to use bleomycin (BLM) combined with monocrotaline (MCT) to induce simulated SSc and the pathogenesis of PAH. The rabbit model of the mechanism is used to explore the relationship between the microcirculation of conjunctiva and PAH, and its early prediction value in the disease. At the same time, vascular endothelial growth factor (VEGF) has been proved to play an important role in microcirculation disturbance and vascular remodeling. This article tries to be in the animal model, PAH, nodal The significance of the membrane microcirculation analysis and the correlation of VEGF level was discussed, and the significance of the spherical conjunctival microcirculation in the SSc combined with PAH was further explored. Methods 1. New Zealand male rabbits were selected and divided into three groups randomly (solvent control group, model group, hormone group), with 8 rats in each group. The model group and hormone group were injected into the abdomen of 1% MCT50mg/kg and daily abdomen respectively. BLM 100ul (concentration of 1mg/ml) was injected subcutaneously, and the hormone group was intraperitoneally injected with methylprednisolone (methylprednisolone, MP) 2.6mg/kg (concentration 1mg/ml) and stopped given three weeks later. Then, the mean pulmonary arterial pressure (mean pulmonary arterial pressure, mPAP) was measured by the right cardiac catheterization, and compared with the solvent control group, the success of the model was judged and passed. The right ventricular hypertrophy index (right ventricular hypertrophy index, RVHI) and the histopathology were perfected to further assess whether the model was consistent with the SSc combined with the pathological features of PAH to analyze the microcirculation of nodular conjunctiva before and after the administration of.2., and the expression of serum VEGF was detected by enzyme linked immunosorbent assay (ELISA). The microcirculation integral of the conjunctiva, VEGF concentration and mPAP were analyzed. The value of the ROC curve was used to determine the value of the microcirculation integral of the bulbar conjunctiva to the diagnosis of SSc combined with PAH. Results: 1. model group mPAP and RVHI were significantly higher than that of the solvent control group and the hormone group. There was a statistically significant (P0.05).2. histopathology: skin staining showed that the epidermis of the skin lesions of the model group was atrophied and hyperkeratosis, and the dermis appeared a small amount of lymph. Cells, plasma cells infiltration, lung tissue staining showed that a large number of bleeding in the alveoli of the model group, small artery myochemical thickening, forming a plexus like structure, accompanied by lymphocyte, plasma cell infiltration.3. conjunctiva microcirculation analysis showed that the total integral of the model group was significantly different from the other two groups (P0.05), mainly manifested in vascular thickening, vascular distortion. In the aspects of reticular malformation, bleeding and blood vessel hue, all the above items were analyzed with the total integral ROC curve of mPAP (P0.05).4. spherical conjunctiva microcirculation. The area under the curve was 0.852 (P0.05). The total score of 5.2 points was the largest, and the total score of the microcirculation of the conjunctiva was more than 5.2 as the demarcation point of the auxiliary diagnostic PAH, and its sensitivity was 88.9. The concentration of serum VEGF in the 75.0%.5. model group was significantly different from that of the other two groups (P0.05), and it was positively correlated with the total integral integral of the conjunctival microcirculation and mPAP (P0.05). Conclusion: 1. the use of BLM combined with MC T injection in New Zealand male rabbits can successfully simulate SSc combined PAH model.2.SSc conjunctival PAH exists the ball conjunctival microcirculation disorder, its severity is There was a positive correlation, especially in vascular thickening, vascular distortion, bleeding, reticular malformation, and blood vessel hue, the total integral of the.3. conjunctiva microcirculation was more than 5.2 points, which could be used as a demarcation point for the diagnosis of PAH, and its sensitivity was 88.9%. The specificity of the degree of specificity was that the serum level of VEGF in the 75.0%.4.SSc combined with PAH increased, and the total score of the conjunctival microcirculation of the ball and mPAP had one. Definite correlation.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.25;R544.1

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