期刊存档
雌激素受体α基因多态性对女性原发性胆汁淤积性肝硬化患者T细胞亚群及其相关细胞因子变化影响的研究
张嫣，陈伟忠，王伟民，王伟民，施杰民张嫣，上海市闸北区中心医院肝胆内科，200070；王伟民，医教科，200070；陈伟忠，第二军医大学附属长征医院消化内科，200070；施杰民，浙江省湖州市中心医院消化内科，200070
摘 要:目的 探讨雌激素受体α(ERα)基因多态性对女性原发性胆汁淤积性肝硬化(PBC)患者T细胞亚群及其相关细胞因子变化的影响。方法 以未经治疗的60例女性PBC患者为研究组，性别、年龄匹配的健康体检的52例为对照组进行研究。采用聚合酶链反应-限制性片段长度多态性方法检测ERα基因1号内含子PvuⅡ和XbaⅠ酶切位点多态性。流式细胞仪定量检测外周血CD4＋、CD8＋、CD4＋CD25＋、CD4＋CD28－ T淋巴细胞。RT-PCR方法检测外周血单个核细胞TNF-α、IL-2、IFN-γ、IL-4、IL-6、IL-10 mRNA的表达。 结果 女性PBC患者的ERα基因PvuⅡ酶切基因亚型中Pp的阳性率明显高于正常对照组(P<0.05)，pp基因亚型的阳性率则明显低于对照组(P<0.05)。XbaⅠ基因型以及等位基因频率在女性PBC组与对照组间差异无统计学意义(P>0.05)。PBC患者的XxPp和xxPp基因型的阳性率均明显高于正常对照组(P<0.05)。PBC患者ppxx基因型的阳性率低于正常对照组，但两组间差异无统计学意义(P>0.05)。女性PBC患者的其他ERα基因型阳性率与正常对照组比较差异均无统计学意义(P>0.05)。PBC患者的T淋巴细胞中CD4＋T淋巴细胞比例增高(P<0.05)，CD8＋T淋巴细胞比例降低(P>0.05)。PBC患者CD4＋CD25＋T细胞比例与对照组比较明显降低(P<0.05)，CD4＋CD28－T细胞比例则明显升高(P<0.05)。PBC患者外周血单个核细胞TNF-α、IL-2和IFN-γ mRNA表达水平明显高于对照组(P<0.01)。IL-4和IL-10 mRNA表达水平虽高于对照组，但组间差异无统计学意义(P>0.05)。IL-6 mRNA表达水平高于对照组(P<0.05)。 结论 PvuⅡ基因的Pp基因型是女性PBC的遗传易感基因型，p等位基因是易感基因。PBC 患者外周血以Th1细胞亚群及其细胞因子占优势。ERα基因多态性可作为遗传易感背景影响PBC患者体内T细胞亚群的偏移及其相关细胞因子的表达。
关键词:肝硬化，胆汁性； 雌激素受体α； 多态现象，遗传； T淋巴细胞； 细胞因子类
Effect of estrogen receptorα gene polymorphism on the variations of T lymphocyte subsets and its related cell factors within female primary cholestasis cirrhosis patient
ZHANG Yan，CHEN Wei-zhong*，WANG Wei-min，SHI Jie-min.*Department of Gastroenterology，Changzheng Hospital，Second Militany Medical University， Shanghai 200003，China
Abstract:Objective To study the effect of estrogen receptor α gene polymorphism on the variations of T lymphocyte subsets and its related cell factors within woman primary cholestasis cirrhosis patient.Methods Untreated female PBC 60 patients were set as study group,52 cases age-matched female for the control.Restriction site polymorphism of Xba Ⅰ and Pvu Ⅱ in ER α gene intron 1 were detected by PCR and restriction fragment length polymorphism.CD4＋，CD8＋，CD4＋CD25＋，CD4＋CD28－T lymphocytes in peripheral blood were quantitatively detected by flow cytometry.RT-PCR was used to detect the expression of TNF-α，IL-2，IFN-γ，IL-4，IL-6，IL-10 in peripheral blood mononuclear cells.Results Positive rate of ERα gene Pvu Ⅱ enzyme gene subtypes Pp with female PBC patients was significantly higher than control (P<0.05).Positive rate of gene pp subtype was significantly lower than control (P<0.05).There was no significant difference (P>0.05) between the PBC group and the control group with genotype Xba I and allele frequency in women.Positive rates of genotype XxPp and xxPp in PBC patients were significantly higher than control (P<0.05).Positive rate of genotypes ppxx in PBC patients was lower than control，whereas no significant difference between the two groups (P>0.05).There was no significant difference between female PBC patients and control with positive rates of other genotypes ERα (P>0.05).The proportion of CD4＋T lymphocytes were increased with PBC patients (P<0.05)，reduced with the proportion of CD8＋T lymphocytes (P>0.05)，increased with the ratio of CD4＋/CD8＋T lymphocytes.The ratio of CD4＋/CD25＋ in PBC patients with was significantly decreased compared with control (P<0.05)，where significantly increased with CD4＋/CD28 (P<0.05).mRNA expression level of TNF-α，IL-2 and IFN-γ in peripheral blood mononuclear cells was significantly higher than that of PBC patients (P<0.01).IL-4，and IL-10 mRNA expression was higher than that of control，whereas no significant (P>0.05).Expression of IL-6 was higher than that of control (P<0.05).Conclusions Pp is a susceptibility genotype of gene Pvu Ⅱ of female PBC，and allele p is a susceptibility gene.Cells subsets Th1 and cytokines are dominant in peripheral blood of PBC patients.As a background of genetic susceptibility，ERα gene polymorphisms can effects T cell subsets alternation and its related cytokines expression in PBC patients.
Key words:Liver cirrhosis，biliary； Estrogen receptor alpha； Polymorphism，genetic； T-lymphocytes； Cytokines
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