子宫内膜异位症患者IL-10基因多态性与血清、腹腔液蛋白水平的相关性研究

目的进一步确定基因多态性是否为子宫内膜异位症(EMs)患者白细胞介素-10(IL-10)蛋白水平改变的原因之一。方法应用扩增阻滞突变系统-聚合酶链反应(ARMS-PCR)结合DNA测序,以及聚合酶链反应-限制性片段长度(PCR-RFLP)多态性分析方法,检测EMs组和对照组IL-10-1082G/A、-819T/C和-592A/C的基因多态性。ELISA法测定血清和腹腔液中IL-10水平。结果 EMs组和对照组均发现3种单倍型GCC、ACC、ATA,6种单倍型组合,其中以ATA/ATA、ACC/ATA和ACC/ACC占多数。对照组仅-1082位点的多态性与血清、腹腔液IL-10水平相关(P0.01)。EMs组-1082(GG或AG型)、-819(CC或TC型)和-592(CC或AC型)均与腹腔液IL-10水平升高相关(P0.05),其中Ⅰ～Ⅱ期EMs患者仅-1082位点与腹腔液IL-10水平相关(P0.01),Ⅲ～Ⅳ期EMs患者-1082、-819和-592位点的基因型均与腹腔液IL-10水平相关(P0.05)。EMs组-1082、-819、-592基因型与血清IL-10水平无相关性(P0.05)。结论Ⅰ～Ⅱ期EMs影响IL-10表达的遗传学机制主要位于-1082位点,但在Ⅲ～Ⅳ期EMs患者中,腹腔局部因素如某些活性因子等除可作用于-1082位点外,可能还可通过作用于-819和-592位点共同调控IL-10水平,但EMs患者血清IL-10水平与其基因多态无相关性,可能与血清IL-10表达受多种机制调控有关。

Research on the Correlations between Interleukin-10 Gene Promoter Polymorphisms and Their Protein Production in Serum and Peritoneal Fluid in Patients with Endometriosis

Objective To investigate whether IL-10 gene promoter polymorphisms are associated with IL-10 protein production in serum and peritoneal fluid and endometriosis risk in Chinese people. Methods Genomic DNA was extracted from blood lymphocytes of 96 Chinese women with different stage EMs and 80 controls. ARMS-PCR （Amplification Refractory Mutation System Polymerase chain reactions） and DNA sequencing were performed to detect polymorphism of-1082G/A site; PCR-RFLP was used to determine the genotypes in-819T/C and-592A/C sites. The serum and peritoneal fluid IL-10 levels were determined by ELISA.Results Three kinds of haplotypes （GCC,ACC and ATA） were found in both groups. Among six kinds haplotypes that have been discovered in present research,ATA/ATA,ATA/ACC and ACC/ACC were the most popular; GCC/ACC and GCC/ATA were less and GCC/GCC was the least. The polymorphism of-1082 site was associated with the serum and peritoneal fluid IL-10 production in controls （P〈0.01）. In Ⅲ~Ⅳ EMs patients the polymorphisms of –1082 （GG or AG genotypes）,–819 （CC or TC genotypes） and –592 （CC or AC genotypes） were associated with the increase of IL-10 levels in peritoneal fluid （P〈0.05）,but inⅠ~ⅡEMs patients only the polymorphism of –1082 was associated with the expression of IL-10 protein in peritoneal fluid. There was no correlation between IL-10 promoter polymorphisms and the serum IL-10 production in EMs group （P〈0.05）. Conclusion The production of IL-10 in controls was mostly regulated by the polymorphism of-1082 site,but in the pathologic conditions,such as endometriosis,not only the polymorphism in-1082 site but also that in-819 and-592 sites may regulate the production of IL-10. There was no correlation between IL-10 promoter polymorphisms and the serum IL-10 production in endometriosis,suggesting that there may be more than one mechanism to regulate the level of IL-10 in serum.