Liang Cuige, DU Wenhua, Dong Qingyu, Liu Xiaomeng, Li Wenxia, Wang Yueli, Gao Guanqi
Department of Endocrinology, Linyi People's Hospital, Linyi, Shandong 276003, P.R. China.
Exp Ther Med. 2015 Mar;9(3):925-930. doi: 10.3892/etm.2015.2180. Epub 2015 Jan 14.
The aim of the present study was to determine whether the expression levels of interleukin (IL)-2 and IL-10 may be used as biological markers in Graves' disease (GD) patients. A total of 256 individuals, including 118 GD patients and 138 healthy individuals, were enrolled into the study. Blood samples were collected from each patient and healthy individual, which were then subjected to enzyme-linked immunosorbent assay (ELISA). Total RNA and total proteins were determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis, respectively. In addition, restriction fragment length polymorphism (RFLP) analysis was performed to detect the presence of genetic polymorphisms. The ELISA results indicated that the IL-2 and IL-10 serum levels in the GD patients were increased by ~5.2 and ~7-fold when compared with the levels in the healthy controls. The results of RT-qPCR indicated that the mRNA expression levels of IL-2 and IL-10 were upregulated in the GD patients when compared with the healthy controls. Furthermore, the western blot analysis results revealed that the protein expression levels of IL-2 and IL-10 were significantly increased in the GD patients. RFLP analysis indicated that the increased number of GG single nucleotide polymorphisms (SNPs) in the GD group were detected in the -330 locus of the IL-2 promoter and the -1082 locus of the IL-10 promoter. In addition, the results indicated that the relatively high rates of homozygous GG SNPs (IL-2 -330T/G and IL-10 -1082A/G polymorphisms) on the alleles may be associated with the incidence of GD. The serum, mRNA and protein expression levels of IL-2 and IL-10 were significantly increased in GD patients when compared with the levels in the healthy controls. In conclusion, the expression levels and genetic polymorphisms of IL-2 and IL-10 may be potential biomarkers for the incidence of Graves' disease in the population studied.
本研究的目的是确定白细胞介素(IL)-2和IL-10的表达水平是否可作为Graves病(GD)患者的生物学标志物。共有256人纳入本研究,其中包括118例GD患者和138例健康个体。从每位患者和健康个体采集血样,然后进行酶联免疫吸附测定(ELISA)。分别使用逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹分析测定总RNA和总蛋白。此外,进行限制性片段长度多态性(RFLP)分析以检测基因多态性的存在。ELISA结果表明,与健康对照相比,GD患者血清中IL-2和IL-10水平分别升高约5.2倍和7倍。RT-qPCR结果表明,与健康对照相比,GD患者中IL-2和IL-10的mRNA表达水平上调。此外,蛋白质印迹分析结果显示,GD患者中IL-2和IL-10的蛋白表达水平显著升高。RFLP分析表明,在IL-2启动子的-330位点和IL-10启动子的-1082位点检测到GD组中GG单核苷酸多态性(SNP)数量增加。此外,结果表明,等位基因上相对较高的纯合GG SNP率(IL-2 -330T/G和IL-10 -1082A/G多态性)可能与GD的发病率相关。与健康对照相比,GD患者中IL-2和IL-10的血清、mRNA和蛋白表达水平显著升高。总之,IL-2和IL-10的表达水平及基因多态性可能是所研究人群中Graves病发病的潜在生物标志物。