Anti-CK18和Anti-CK19自身抗体与慢性阻塞性肺疾病的相关性研究

发布时间：2018-05-24 22:03

本文选题：COPD + 自身抗体　；参考：《湖北中医药大学》2017年硕士论文

【摘要】：目的:近年来,许多研究报道,自身免疫在慢性阻塞性肺疾病(COPD)的发生发展中起重要作用,而且针对CK18和CK19的自身抗体与肺组织损伤有关。本研究通过检测COPD患者血浆中anti-CK18和anti-CK19IgG、IgA、IgM型自身抗体水平变化以及研究香烟烟雾对CK18和CK19的自身抗体产生的影响,探讨anti-CK18和anti-CK19自身抗体在COPD发生发展中的作用。方法:1.收集COPD患者血标本228例和年龄性别匹配的正常对照血标本136例,用间接酶联免疫吸附试验检测血浆中anti-CK18和anti-CK19 IgG、IgA、IgM型自身抗体的水平变化,分析anti-CK18和anti-CK19自身抗体与COPD患者肺功能不同分期的关系,同时对不同分期COPD患者的临床资料进行分析;2.C57BL/6J雄性小鼠暴露于香烟烟雾中建立COPD动物熏烟模型,分别暴露6个月和9个月,同时设立正常对照组,取其血清和肺脏,应用间接酶联免疫吸附试验检测香烟烟雾暴露组和正常对照组小鼠血清中anti-CK18和anti-CK19 IgG、IgA、IgM型自身抗体的水平变化;3.用4%多聚甲醛固定小鼠左肺,HE染色检测经香烟烟雾暴露后小鼠肺组织的病理学改变;用间接免疫荧光检测香烟烟雾暴露组和正常对照组小鼠肺组织中的IgG、IgA、IgM型抗体分布情况;4采用western blot技术检测香烟烟雾暴露组小鼠肺组织中是否存在anti-CK18和anti-CK19 IgG型自身抗体。结果:1.COPD患者血浆anti-CK18和anti-CK19 IgG、IgA、IgM型自身抗体水平明显高于正常对照组,尤其是COPD III期和GOLD IV期,差异具有统计学意义(p0.05)。随着疾病的严重程度逐渐增加,anti-CK18 IgG、IgM型自身抗体和anti-CK19 IgA、IgM型自身抗体的水平逐渐增加,但是anti-CK18 IgA型抗体和anti-CK19 IgG型抗体水平并未随着疾病加重而逐渐增加。COPD患者具有明显的吸烟史趋势,其肺功能较正常对照组显著降低。此外,anti-CK18 IgG、IgA、IgM型自身抗体和anti-CK19 IgA、IgM型自身抗体与COPD患者肺功能FEV1占预计值百分比之间呈负相关;2.6个月和9个月香烟烟雾暴露组小鼠血清anti-CK18和anti-CK19 IgG型抗体水平明显较对照组高,但是血清anti-CK18和anti-CK19 IgG型抗体在6个月与9个月烟雾暴露组之间并没有明显变化,说明其IgG型抗体水平变化与烟雾暴露环境密切相关,差异具有统计学意义(p0.05)。而在香烟烟雾暴露9个月后小鼠血清anti-CK18和anti-CK19 IgA型抗体水平显著高于香烟烟雾暴露6个月后小鼠血清含量;3.香烟烟雾暴露组小鼠的肺组织出现明显的病理改变,香烟烟雾暴露6个月后小鼠肺泡壁出现轻度破坏,伴有大量炎性细胞浸润,香烟烟雾暴露9个月后小鼠肺组织出现大量的肺泡融合扩大,并伴有部分炎性细胞浸润;小鼠肺部IgG、IgA、IgM型抗体主要分布于肺泡间隔、小气道壁和血管壁。6个月和9个月香烟烟雾暴露组小鼠肺组织的IgG、IgA、IgM型抗体表达率比较分别为57.1%、42.9%、57.1%和40%、33.3%、46.7%,表达量较对照组显著升高;4.Western blot检测结果显示在香烟烟雾暴露组小鼠IgG、IgA、IgM型抗体分布阳性的肺组织中存在针对CK18和CK19的IgG型自身抗体。结论:Anti-CK18和anti-CK19 IgG、IgA、IgM型自身抗体是疾病相关自身抗体,其循环水平与COPD疾病的严重程度密切相关,在监测COPD疾病发展中起到重要作用,有望作为监测COPD肺组织损伤严重程度的标志物,进一步指导COPD疾病的治疗和预后。
[Abstract]:Objective: in recent years, many studies have reported that autoimmunity plays an important role in the development of chronic obstructive pulmonary disease (COPD), and the autoantibodies against CK18 and CK19 are related to lung tissue damage. This study was conducted to detect the changes in the level of anti-CK18 and anti-CK19IgG, IgA, IgM type autoantibodies and the study of cigarette smoke in the plasma of COPD patients. The effect of fog on the production of autoantibodies of CK18 and CK19 and the role of anti-CK18 and anti-CK19 autoantibodies in the development of COPD. Methods: 1. collect 228 blood samples from COPD patients and 136 normal control blood samples matched by age and sex, and detect anti-CK18 and anti-CK19 IgG, IgA, IgM type in plasma by indirect enzyme-linked immunosorbent assay. The change of the body antibody level, the relationship between the anti-CK18 and anti-CK19 autoantibodies and the different stages of the pulmonary function in COPD patients, and the analysis of the clinical data of the patients with different stages of COPD; the 2.C57BL/6J male mice were exposed to cigarette smoke to establish the COPD smoked smoke model for 6 months and 9 months, and the normal control group was set up at the same time. The levels of anti-CK18 and anti-CK19 IgG, IgA, IgM type autoantibodies in the serum of cigarette smoke exposure group and normal control group were detected by indirect enzyme linked immunosorbent assay (ELISA). 3. the left lung of mice was fixed with 4% polyformaldehyde, and the pathological changes of lung tissue in mice after exposure to smoke smoke were detected by HE staining; The distribution of IgG, IgA and IgM type antibodies in lung tissue of cigarette smoke exposure group and normal control group was detected by immunofluorescence. 4 the presence of anti-CK18 and anti-CK19 IgG autoantibodies in lung tissue of cigarette smoke exposed mice was detected by Western blot. The level of autoantibody was significantly higher than that in the normal control group, especially in the COPD III phase and GOLD IV phase. The difference was statistically significant (P0.05). As the severity of the disease gradually increased, the level of anti-CK18 IgG, IgM type autoantibody and anti-CK19 IgA, IgM type autoantibody gradually increased. The level of.COPD patients did not gradually increase with the increase of the disease. Their lung function was significantly lower than that in the normal control group. In addition, anti-CK18 IgG, IgA, IgM type autoantibodies and anti-CK19 IgA, IgM type autoantibodies were negatively correlated with the percentage of lung function FEV1 in COPD patients; 2.6 months and 9 months of cigarettes The levels of serum anti-CK18 and anti-CK19 IgG antibodies in the smoke exposure group were significantly higher than those in the control group, but there was no significant change between the serum anti-CK18 and anti-CK19 IgG antibodies between 6 months and 9 months of smoke exposure, indicating that the changes in the level of IgG type antibodies were closely related to the environment of the smoke exposure, and the difference was statistically significant (P0.05). After 9 months of exposure to cigarette smoke, the level of anti-CK18 and anti-CK19 IgA antibodies in mice was significantly higher than that of the mice exposed to cigarette smoke for 6 months. The lung tissues of the mice exposed to cigarette smoke exposed a significant pathological change. After 6 months of cigarette smoke exposure, the alveolar walls of mice were slightly damaged, accompanied by a large number of inflammatory fines. After 9 months of exposure to cigarette smoke, a large number of pulmonary alveolar fusion and inflammatory cell infiltration were found in the lung tissue of mice. The IgG, IgA, and IgM type antibodies in the lungs of mice were mainly distributed in the alveolar septum, and the expression rates of IgG, IgA and IgM antibodies in the lung tissues of the mice of the small air channel wall and the vessel wall for the.6 months and the 9 months of cigarette smoke exposure group were compared. 57.1%, 42.9%, 57.1% and 40%, 33.3%, 46.7%, respectively, the expression levels were significantly higher than those in the control group. 4.Western blot detection showed that the IgG, IgA, IgM type antibody in the lung tissues of the mice exposed to cigarette smoke exposed the IgG type autoantibodies against CK18 and CK19. Conclusion: Anti-CK18 and anti-CK19 IgG, IgA, and autoimmune antibodies are disease. Related autoantibodies, which are closely related to the severity of COPD disease, play an important role in monitoring the development of COPD disease, and are expected to be a marker for monitoring the severity of COPD lung tissue damage and further guide the treatment and prognosis of COPD disease.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R563.9

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