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目的研究颗粒溶素(granulysin,GNLY)、颗粒酶 B(granzyme B,GrB)及穿孔素(perforin,PF)三种免疫效应分子在原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)外周血中的表达并探讨其与 PBC 的关系。方法以正常健康人和肝炎后肝硬化患者为对照,用实时荧光定量 RT-PCR 方法检测 PBC 患者 PBMC 中 GNLY、GrB 及 PF mRNA 的含量。用 ELISA 方法检测 PBC 患者血清中 GNLY 表达情况并分析其与肝功能的相关性。结果 PBC 组 GNLY、GrB mRNA 平均拷贝数显著高于健康对照组(P<0.01)和疾病对照肝炎后肝硬化组(P<0.001),而 PF mRNA 与对照组相比差异无统计学意义。PBC 患者血清中的 GNLY 蛋白表达显著高于健康对照组(P<0.01)和乙型肝炎后肝硬化组(P<0.01)。血清中 GNLY 浓度与血清 GGT、ALP 的浓度呈正相关(P<0.01)。结论 PBC 患者 GNLY、GrB mRNA 和含量显著高于对照组,同时 ELISA 方法检测出 PBC 患者血清中 GNLY 蛋白含量显著高于对照组,GNLY、GrB 表达与 PBC 的发病存在一定的关联性,为临床 PBC 病情的监控提供可靠依据。
Objective To study the effect of granulelin (GNLY), granzyme B (GrB) and perforin (PF) on peripheral blood of primary biliary cirrhosis (PBC) In the expression and explore its relationship with PBC. Methods The normal and healthy post-hepatitis cirrhosis patients were used as control. The contents of GNLY, GrB and PF mRNA in PBMC of PBC patients were detected by real-time fluorescence quantitative RT-PCR. The serum levels of GNLY in PBC patients were detected by ELISA and their correlation with liver function was analyzed. Results The average copy number of GNLY and GrB mRNA in PBC group was significantly higher than that in healthy control group (P <0.01) and disease control cirrhosis group (P <0.001). However, there was no significant difference between PF mRNA and control group. The GNLY protein expression in serum of PBC patients was significantly higher than that of healthy controls (P <0.01) and posthepatitic cirrhosis (P <0.01). Serum GNLY concentration and serum GGT, ALP concentration was positively correlated (P <0.01). Conclusions The mRNA and protein levels of GNLY and GrB in PBC patients were significantly higher than those in control group. Meanwhile, the serum levels of GNLY protein in PBC patients were significantly higher than those in control group. The expression of GNLY and GrB was correlated with the incidence of PBC. Disease monitoring to provide a reliable basis.