【摘要】：目的:原發(fā)性干燥綜合征(Primary Sj?gren’s Syndrome,p SS)是以侵犯唾液腺、淚腺等外分泌腺體,以大量淋巴細(xì)胞浸潤(rùn)為特征的自身免疫性疾病。多項(xiàng)研究發(fā)現(xiàn),p SS的發(fā)病與EB病毒(Epstein-Barr virus,EBV)的感染相關(guān)。EB病毒核抗原-1(Epstein-Barr Nuclear Antigen-1,EBNA-1)和EB病毒核抗原-2(Epstein-Barr Nuclear Antigen-2,EBNA-2)是EBV潛伏期表達(dá)的核心抗原。通過(guò)檢測(cè)p SS患者外周血單個(gè)核細(xì)胞(Peripheral blood mononuclear cell,PBMC)EBV DNA拷貝數(shù),以及測(cè)定p SS患者唇腺組織中EBNA-1和EBNA-2的表達(dá)情況,探討EBV與p SS發(fā)病的關(guān)系。方法:應(yīng)用Real-time熒光定量PCR定量檢測(cè)32例p SS患者與28例正常對(duì)照PBMC中EBV DNA拷貝數(shù)。通過(guò)免疫組化的方法測(cè)定32例p SS患者及6例正常唇腺組織中EBNA-1和EBNA-2的表達(dá)情況(以細(xì)胞胞漿、胞核中出現(xiàn)棕黃色顆粒狀物質(zhì)為陽(yáng)性)。結(jié)果:(1)p SS患者EB病毒DNA檢測(cè)陽(yáng)性率100%(32/32),對(duì)照組EB病毒DNA檢測(cè)陽(yáng)性率89.2%(25/28),經(jīng)檢驗(yàn),差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(2)32例p SS患者EB病毒DNA拷貝數(shù)均值為26.2±10.85 copies/μg,28例對(duì)照組EB病毒DNA拷貝數(shù)均值為8.6±5.21 copies/μg,經(jīng)檢驗(yàn),患者組EB病毒DNA拷貝數(shù)均值明顯高于對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。(3)32例p SS患者臨床表現(xiàn)與EB病毒DNA拷貝數(shù)比較顯示,腮腺腫大組較非腮腺腫大組EB病毒DNA拷貝數(shù)差異有統(tǒng)計(jì)學(xué)意義(P0.05)。32例p SS患者實(shí)驗(yàn)室檢查結(jié)果與EBV-DNA之間的相關(guān)性分析顯示,Ig G與病毒DNA拷貝數(shù)呈正相關(guān)(p0.05)。(4)EBNA-1和EBNA-2在部分p SS患者唇腺組織浸潤(rùn)的淋巴細(xì)胞,唇腺導(dǎo)管上皮細(xì)胞中可見(jiàn)陽(yáng)性表達(dá),而對(duì)照組唇腺組織未見(jiàn)陽(yáng)性表達(dá)。p SS唇腺組織中EBNA-1的陽(yáng)性率為68.8%(22/32),EBNA-2的陽(yáng)性表達(dá)率62.5%(20/32),較正常對(duì)照組比較均有顯著統(tǒng)計(jì)學(xué)意義(P0.05)。(5)EBNA-1在p SS唇腺組織中陽(yáng)性和陰性的患者在臨床表現(xiàn)和實(shí)驗(yàn)室檢查方面無(wú)明顯差異性(P0.05)。EBNA-2在p SS唇腺組織中陽(yáng)性和陰性的患者在臨床表現(xiàn)和實(shí)驗(yàn)室檢查方面無(wú)明顯差異性(P0.05)。結(jié)論:(1)EBV復(fù)制可能參與p SS的發(fā)病。(2)p SS腮腺腫大的患者提示可能EBV復(fù)制活躍,加重發(fā)病。(3)高Ig G提示與EBV復(fù)制活躍相關(guān)。(4)EBNA-1、EBNA-2可能通過(guò)參與EBV感染而與p SS的發(fā)病相關(guān)。
[Abstract]:Objective: primary Sjgren's Syndromeg syndrome (SS) is an autoimmune disease characterized by invasion of exocrine glands such as salivary glands and lacrimal glands, and massive lymphocytic infiltration. Many studies have found that Epstein-Barr virus (EBV) infection and Epstein-Barr Nuclear Antigen-1EBNA-1 (EBNA-1) and Epstein-Barr Nuclear Antigen-2( EBNA-2) are the core antigens of EBV expression. The expression of EBNA-1 and EBNA-2 in labial gland of patients with PSS was determined by detecting the copy number of EBV DNA in peripheral blood mononuclear cells (PBMCs) of patients with PSS, and the relationship between EBV and PSS was studied. Methods: Real-time quantitative PCR was used to detect EBV DNA copy number in PBMC of 32 patients with PSS and 28 normal controls. The expression of EBNA-1 and EBNA-2 in 32 cases of PSS and 6 cases of normal labial gland were detected by immunohistochemical method. Results: (1) Epstein-Barr virus DNA positive rate was 100% (32 / 32) in patients with PSS and 89.2% (25 / 28) in controls. There was no significant difference (P0.05). (2). The average copy number of EBV DNA in 32 patients with PSS was 26.2 鹵10.85 copies/ 渭 g. The average value of EBV DNA copy number in 28 cases of control group was 8.6 鹵5.21 copies/ 渭 g, and the average value of EBV copy number in patients with PSS was significantly higher than that in control group. The difference was statistically significant (P0.05). (3). The clinical manifestations of 32 patients with PSS were compared with Epstein-Barr virus copy number (EBV). Epstein-Barr virus DNA copy number in parotid enlargement group was significantly different from that in non-parotid enlargement group (P0.05). The correlation analysis between EBV-DNA and EBV-DNA in 32 patients with PSS showed that Ig was positively correlated with viral DNA copy number (p0.05). (_ 4) EBNA-1 and EBNA-2 in the region. Lymphocytes infiltrated in labial gland of patients with PSS, Positive expression was found in epithelial cells of labial gland ducts. The positive rate of EBNA-1 was 62.8% (22 / 32) in the labial gland of the control group, which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in the labial gland tissue of PSS was 62.5% (20 / 32), which was significantly higher than that in the control group (P0.05). The positive rate of EBNA-1 in PSS labial gland was significantly higher than that in the control group (P0.05). There was no significant difference between clinical manifestation and laboratory examination (P0.05). There was no significant difference in clinical manifestation and laboratory examination between patients with EBNA-2 positive and negative in PSS labial gland (P0.05). Conclusion: (1) EBV replication may be involved in the pathogenesis of PSS. (2) the patients with PSS parotid gland enlargement may be active in EBV replication and aggravate the disease. (3) the high IgG hint may be related to EBV replication activity. (4) EBNA-1 EBNA-2 may be associated with the pathogenesis of PSS by participating in EBV infection.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R593.2

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