【摘要】 目的观察孕激素对高迁移率组蛋白B1(HMGB1)诱导人脐静脉内皮细胞(HUNVEC)释放细胞因子白细胞介素-6(IL-6)的影响。方法分离培养人原代HUVEC,克隆构建HMGB1原核重组表达载体pET14b-HMGB1;用不同浓度的原核表达纯化的HMGB1(0、10、100、500、1000ng/ml)蛋白和不同浓度的孕酮(0、0.1、1、10、100mmol/L)刺激HUVEC,24h后用酶联免疫吸附法(ELISA)检测细胞上清中IL-6的表达水平;用500ng/ml的HMGB1分别与不同浓度的孕酮联合作用刺激培养的HUVEC,ELISA法检测细胞上清中IL-6的表达水平;分析孕酮对HMGB1诱导IL-6影响的剂量效应。结果不同浓度的HMGB1刺激HUVEC后发现,低浓度HMGB1可轻度下调IL-6的分泌表达,但无明显统计学差异(P>0.05),一定浓度以上则可使IL-6水平明显升高(P<0.01);不同浓度孕激素对HUVEC产生IL-6没有明显影响,但以剂量依赖方式抑制HMGB1对IL-6的分泌(P<0.01)。结论HMGB1可诱导HUVEC释放炎性细胞因子IL-6,而孕激素以剂量依赖的方式抑制这些炎症因子的释放,这为孕激素在内毒素血症的发生发展中发挥保护作用提供了分子水平的理论基础。更多还原

【Abstract】 Objective To study the effect of progesterone on interleukin-6 (IL-6) release from human umbilic vein endothelial cells (HUVECs) induced by high mobility group box-1 protein (HMGB1). Methods The recombinant expression plasmid pET14b-HMGB1 was constructed and transformed into competent E.coli BL21 cells to obtain HMGB1 protein, which was purified with chromatography on Ni-NTA Sepharose column. Cultured HUVECs were treated with purified HMGB1 protein alone at the concentrations 0, 10, 100, 500, and 1000 ng/ml, progesterone alone at the concentrations of 0, 0.1, 1, 10, 100 mmol/L, or with both HMGB1 protein (500 ng/ml) and progesterone at the terminal concentrations of 0, 0.1, 1, 10, and 100 mmol/L. Twenty-four hours later, the supernatant of the cell culture medium was collected to detect the levels of IL-6 using enzyme-linked immunosorbent assay (ELISA). Results The IL-6 levels in HUVEC culture medium was slightly decreased after treatment with low-concentration HMGB1 but increased obviously following treatment with high-concentration HMGB1, and these effects could be dose-dependently inhibited by progesterone. Progesterone alone did not result in any noticeable changes of IL-6 levels in the cell culture medium. Conclusion Progesterone can dose-dependently inhibit HMGB1-induced IL-6 release from HUVECs, suggesting the protective role of progesterone in endotoxemia.更多还原