急性肺血栓栓塞动物模型的建立与IL-13、TGF-β在肺栓塞中的作用初步探讨
发布时间：2018-02-07 13:29
本文关键词： 急性肺血栓栓塞 肺动脉高压 IL-13 TGF-β 动物模型 出处：《天津医科大学》2011年硕士论文 论文类型：学位论文
【摘要】：目的：肺血栓栓塞症(Pulmonary thromboembolism, PTE)为来自静脉系统或右心的血栓阻塞肺动脉或其分支所致的疾病,以肺循环和呼吸功能障碍为主要临床和病理生理特征。目前对于PTE在分子及细胞水平的病理生理了解尚少,多数学者认为肺栓塞的病理进程与炎症反应有关。最近有文献报道了肺动脉重建中的免疫学发病机理。本文提出假设认为PTE后肺动脉重建与炎症反应及免疫反应相关,并进行论证。 IL-13是由不同T细胞亚群和树突状细胞产生的多效能细胞因子,主要由Th2细胞分泌。有文献报道IL-13系统是肺动脉平滑肌细胞生长的新型调节系统。 此外,IL-13可以诱导潜在TGF-β的产生,间接激活TGF-β。同时,低氧也可促进TGF-β表达增高。而TGF-β是调控细胞增殖、分化的重要因子,能促进血管平滑肌细胞增殖、表型转化及血管重构,与多种心血管疾病发生密切相关。 本课题诣在建立急性肺血栓栓塞模型,观察肺栓塞的病理改变,并探讨IL-13、TGF-β在PTE肺动脉重建中的作用。 方法：本课题分三部分进行： 第一部分：日本大耳白36只,随机分为急性肺血栓栓塞模型组18只、对照组18只。模型组采用自体血栓回输法建立PTE模型,对照组给予等量0.9%氯化钠溶液,经数字减影血管造影及病理学检查确定模型建造成功与否。 第二部分：分别于48h、4d、14d采集模型组和对照组肺动脉及周围肺组织,行HE染色及Masson染色进行病理学分析。 第三部分：分别于48h、4d、14d采集模型组及对照组血清,应用酶联免疫吸附试验方法(ELISA)检测IL-13、TGF-β含量。 实验数据采用SPSS 13.0统计软件进行统计学分析,数据以均数±标准差表示,组内不同时间点比较采用方差分析(One-Way ANOVA),各组间均数比较采用t检验(t-test analysis),数据相关性分析采用Pearson检验,均以P0.05为差异有统计学意义。 结果：第一部分：对22只实验兔进行造模,成功制作18只PTE模型。未成功的4只动物中,1只死于麻醉意外,3只死于大面积栓塞,成功率为81.8%。栓塞水平多位于段肺动脉,以双肺栓塞为主,下肺为著。RR、HR、AaDO2升高均发生于肺栓塞早期。 第二部分：受累肺动脉壁及远端肺动脉分别于第4天和第2天可见以中性粒细胞为主的炎症细胞数量增加(P0.05)；于栓塞后第14天可见肺动脉内膜明显增厚(P0.05) 第三部分：肺栓塞后血清中IL-13、TGF-B水平分别于第4天和第14天升高,为36.86±3.06pg/mg、1591.67±55.01pg/mg,显著高于对照组(P0.05)。结论：1.自体血栓回输法制备急性肺动脉血栓栓塞动物模型成功率为81.8%,具有可重复性。栓子分布与临床上栓子分布相似。AaDO2可作为肺栓塞的早期诊断指标。RR及HR变化于肺栓塞后即刻出现,但HR的恢复早于RR。 2.肺栓塞中,炎症细胞浸润不仅发生在受累肺动脉壁,也发生在远端肺动脉,均以中性粒细胞为主；肺泡间隔内可见以中性粒细胞为主的大量炎性细胞浸润,可见肺泡膨胀不全。阐明了PTE的病理学改变与炎症细胞浸润相关。 3.在肺栓塞进程中Th2型免疫反应占有一定地位,IL-13与TGF-B共同促进胶原产生,导致纤维化,参与肺动脉重建。但两个指标并无相关性。
[Abstract]:Objective: pulmonary thromboembolism (Pulmonary thromboembolism, PTE) from the venous system or right heart thromboembolism of pulmonary artery or its branches caused by the disease, pulmonary circulation and respiratory dysfunction as the main clinical and pathophysiological features. The PTE in the understanding of the pathophysiology of molecular and cellular level is less, the majority of scholars believe that the pathological process of inflammation and pulmonary embolism. Recently reported the immunological pathogenesis of pulmonary artery reconstruction. This paper proposed a hypothesis that related to pulmonary artery reconstruction and inflammation after PTE and immune response, and demonstrate.
IL-13 is a multipotent cytokine produced by different T cell subsets and dendritic cells, mainly secreted by Th2 cells. It has been reported that IL-13 system is a new regulation system for the growth of pulmonary artery smooth muscle cells.
In addition, IL-13 can induce the potential of TGF- beta, indirect activation of TGF- beta. At the same time, hypoxia may also promote the expression of TGF- increased. TGF- beta is an important factor in the regulation of cell proliferation, differentiation, can promote vascular smooth muscle cell proliferation, phenotype transformation and vascular remodeling, is closely related with various cardiovascular diseases.
This paper aims to establish models of acute pulmonary embolism, observe the pathological changes of pulmonary embolism, and to explore the role of IL-13, TGF- in beta PTE pulmonary artery reconstruction.
Methods: the subject was divided into three parts:
The first part: Japanese big ear white 36 rabbits were randomly divided into the model of acute pulmonary embolism in group 18, 18 rats in the control group. The model group using autologous blood transfusion method to establish the PTE model, the control group received the same amount of 0.9% Sodium Chloride Solution, by digital subtraction angiography and pathological examination to determine the model construction is successful or not.
The second part: the pulmonary artery and peripheral lung tissue were collected in 48h, 4D, 14d and control group respectively. The pathological analysis was performed by HE staining and Masson staining.
The third part: the serum of the model group and the control group were collected in 48h, 4D, 14d, and the content of IL-13 and TGF- beta was detected by enzyme linked immunosorbent assay (ELISA).
The experimental data were analyzed by SPSS 13 statistical software, data with the mean standard deviation of that group at different time points in comparison with analysis of variance (One-Way, ANOVA) among the groups were compared using t test (t-test analysis), the correlation analysis of data using Pearson test, P0.05 was considered statistically significant.
缁撴灉锛氱涓，
本文编号：1494485
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