原发性胆汁性胆管炎患者唾液中抗线粒体抗体检测及口腔免疫环境变化的研究

发布时间：2018-06-07 07:34

本文选题：原发性胆汁性胆管炎 + 抗线粒体抗体　；参考：《浙江大学》2017年硕士论文

【摘要】：研究背景及目的:原发性胆汁性胆管炎(primary biliary cholangitis,PBC)是一种进行性的自身免疫性肝病,主要引起患者肝内胆管损伤,引发胆汁淤积性症状,导致肝纤维化,最终导致肝衰竭。PBC的发病机制尚不明确,PBC的发病机制尚不明确,并无特异性的临床表现,早期诊断率并不是很高,而PBC晚期的患者只能进行肝移植治疗。因此,对高危人群进行定期筛查,提高PBC的早期诊断率,对PBC患者的治疗以及改善患者的预后均有很大的帮助。抗线粒体抗体M2亚型(AMA-M2)是临床上诊断PBC的特异性生物学指标,临床上主要通过血清途径进行AMA-M2的检测,但是,血清检测途径具有很多自身缺陷,如有创伤、感染风险、不够方便等,因此,AMA-M2的无创检测方法具有很大的研究前景。本实验目的在于利用自主改进的方法检测唾液中AMA-M2的水平及唾液中炎症因子的表达,评估唾液中AMA-M2在该疾病中的诊断价值,以及PBC患者口腔免疫环境的变化,进而对PBC的早期诊断提供新的方法。研究方法本研究收集了 PBC患者、口腔扁平苔藓(oral lichen planus,OLP)患者和健康成年人(health controls,HC)的血清和唾液样本共151例。利用酶联免疫吸附法(Enzyme-linked Immunosorbent assay,ELISA)检测三组血清和唾液样本中AMA-M2的表达水平,用液相芯片检测三组唾液和血清样本中炎症性细胞因子的表达水平,通过t检验,单因素方差分析比较分析各组AMA-M2表达水平及细胞因子表达水平之间的差异,还利用Pearson相关性分析的方法分析了 AMA-M2及细胞因子表达水平在唾液和血液之间的联系。同时,我们还检测了 PBC组和HC组血清中碱性磷酸酶(Alkaline phosphatase,ALP)、谷氨酰转肽酶(Gamma-glutamyl transpeptidase,GGT)的水平。研究结果:研究发现唾液的AMA-M2水平只能在血清AMA-M2表达阳性的PBC患者中测得,而在血清AMA-M2表达阴性的PBC患者、OLP组、HC组中,唾液AMA-M2的表达低于检测下限。同时,我们发现在血清AMA-M2表达阳性的PBC患者中,血清和唾液中AMA-M2表达水平存在明显的正相关性。另一方面,在PBC组和OLP组唾液中IL-6、IFN-γ,TNF-α,IL-17A表达水平均比HC组明显升高,而IL-10的变化不明显,并且OLP组唾液中炎症性细胞因子表达水平比PBC组也有明显升高。结论:唾液具有无创、便捷的优势,且能够一定程度上反映血液中相关成分的表达情况。唾液AMA-M2在PBC诊断中有较高的灵敏度和特异度,在PBC的早期诊断和高危群体的筛查中具有较高的应用价值,也为PBC的早期诊断提供了一个新的方向。同时,我们发现PBC患者口腔内存在Th1/Th2失衡,Th17相关炎性细胞.因子的表达升高,患者发病过程中可能存在着出现口腔炎症性疾病风险,这为临床上PBC的治疗提供了更多的思路与理论基础。
[Abstract]:Background and objective: primary biliary cholangitis is a progressive autoimmune liver disease. The pathogenesis of liver failure. PBC is not clear, there is no specific clinical manifestation, the early diagnosis rate is not very high, but the late PBC patients can only be treated with liver transplantation. Therefore, regular screening of high-risk groups to improve the rate of early diagnosis of PBC, the treatment of PBC patients and improve the prognosis of patients have a great help. AMA-M2) is a specific biological index for clinical diagnosis of PBC. In clinic, AMA-M2 is mainly detected by serum pathway. However, serum detection pathway has many defects, such as trauma and risk of infection. Therefore, the non-invasive detection method of AMA-M2 has great research prospect. The purpose of this study was to detect the level of AMA-M2 in saliva and the expression of inflammatory factors in saliva, to evaluate the diagnostic value of AMA-M2 in saliva and the changes of oral immune environment in patients with PBC. It provides a new method for early diagnosis of PBC. Methods Serum and saliva samples of PBC patients, oral lichen planus lichen patients and healthy adults were collected. Enzyme linked immunosorbent assay (Elisa) was used to detect the expression of AMA-M2 in serum and saliva samples of three groups, and the expression of inflammatory cytokines in saliva and serum samples of three groups was detected by liquid-phase microarray, and the expression of inflammatory cytokines in saliva and serum samples was detected by t test. Univariate ANOVA was used to compare and analyze the differences of AMA-M2 expression and cytokine expression in each group. The relationship between saliva and blood of AMA-M2 and cytokine expression was also analyzed by Pearson correlation analysis. At the same time, we also detected the levels of alkaline phosphatase (ALP) and glutamyl transpeptidase (GGTs) in the serum of PBC and HC groups. Results: the results showed that the AMA-M2 level of saliva could only be detected in PBC patients with positive serum AMA-M2 expression, but the expression of AMA-M2 in saliva was lower than the lower detection limit in PBC patients with negative AMA-M2 expression. At the same time, we found that there was a positive correlation between AMA-M2 expression in serum and saliva in PBC patients with positive AMA-M2 expression. On the other hand, the level of IL-6 IFN- 纬 TNF- 伪 IL-17A in saliva of PBC group and OLP group was significantly higher than that of HC group, but the change of IL-10 was not obvious, and the expression of inflammatory cytokines in saliva of OLP group was significantly higher than that of PBC group. Conclusion: saliva has the advantages of noninvasive and convenient, and can reflect the expression of related components in blood to some extent. Salivary AMA-M2 has high sensitivity and specificity in the diagnosis of PBC. It has a high value in the early diagnosis of PBC and screening of high-risk population. It also provides a new direction for the early diagnosis of PBC. At the same time, we found that Th1/Th2 imbalance and Th17-related inflammatory cells were found in the oral cavity of PBC patients. The increased expression of factors may lead to the risk of oral inflammatory diseases in the pathogenesis of patients, which provides more ideas and theoretical basis for the clinical treatment of PBC.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.7

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