原发性胆汁性肝硬化患者B淋巴细胞刺激因子基因表达的测定及临床意义  

Determination and clinical significance of B-lymphocyte stimulator gene expression on peripheral blood mononuclear cells from patients with primary biliary cirrhosis

作　　者：梁艳[1] 杨再兴[1] 周琳[1] 朱烨[1] 陈燕[1] 邓安梅[1] 仲人前[1] 

机构地区：[1]第二军医大学长征医院实验诊断科全军临床免疫中心,上海200003

出　　处：《检验医学》2007年第1期38-40,共3页Laboratory Medicine

基　　金：国家自然科学基金资助项目(30080027);上海市科研计划项目(04DZ19116;051409012)

摘　　要：目的建立检测人B淋巴细胞刺激因子(BLyS)mRNA的相对定量方法,探讨其基因表达水平与原发性胆汁性肝硬化(PBC)的相关性。方法基于TaqM an-MGB探针技术,以18SrRNA为内参照,通过目的基因与内参基因荧光强度达到一定阈值时的循环数之差(△CT值)定量原始模板浓度,建立实时荧光逆转录聚合酶链反应(FQ-RT-PCR)相对定量方法,检测30名正常人和40例PBC患者外周血单个核细胞(PBMC)BLyS基因的表达。结果PBC患者外周血单个核细胞BLyS mRNA表达的△CT值为9.5±2.5,与正常对照组比较,差异有统计学意义(P<0.01),其原始模板浓度为正常对照组的4.3倍。PBC患者中抗核抗体(ANA)阳性占80%;其中ANA 1∶100组BLyS检测的△CT为11.4±0.9,与正常对照组比较,差异无统计学意义(P>0.05);而ANA 1∶1 000和ANA 1∶10 000组BLyS的△CT值与正常对照组比较,差异有统计学意义(P<0.001),两组间△CT值比较差异有统计学意义(P<0.01)。结论PBC患者PBMC BLyS mRNA表达水平明显升高,且与ANA滴度相关。Objective To establish fluorescent semi-quantitative polymerase chain reaction （PCR） for detecting human B-lymphocyte stimulator （BLyS） mRNA and to explore the association of its expression level with primary biliary cirrhosis（PBC）. Methods Based on fluorescent TaqMan-MGB technology, with 18SrRNA as control, real-time fluorescent semi-quantitative reverse transcriptional PCR was established, according to the difference of threshold cycles （△CT） of BLyS and 18SrRNA to determine the original templates. BLyS mRNA of peripheral blood mononuclear cells （PBMC） in 30 healthy people and 40 PBC patients was measured. Results BLyS mRNA level was significantly higher in patients with PBC than in control（ P 〈 0.01 ）. The concentration of the original template of BLyS gene in PBC patients was 4.3 times as much as that of control. 80% of PBC patients were positive for antinuclear antibody（ANA） ;△CT of BLyS in ANA 1 ： 100 group was 11.4 ± 0.9, showing no significant difference from that of control （ P 〉 0.05 ）. Comparatively, △CTs of ANA 1：1 000 and ANA 1：10 000 groups were significantly different from that of control. Conclusions BLyS gene expression level of PBMC in PBC patients elevates significantly, which is associated with ANA titers.

关 键 词：B淋巴细胞刺激因子 原发性胆汁性肝硬化 聚合酶链反应 

分 类 号：Q503[生物学—生物化学]