摘要
目的探讨雌二醇在骨关节炎病程中对滑膜细胞产生抗炎作用的机制。方法分离并鉴定滑膜细胞,利用免疫荧光染色观察滑膜细胞表达雌激素受体β(ERβ)的情况;而后对滑膜细胞进行分组:空白对照组,10 ng/m L白细胞介素1β(IL-1β)处理组、雌二醇处理组(用10 ng/m L IL-1β预处理同时,添加10-7mol/L的雌二醇培养)、雌二醇联合雌激素受体阻断剂四氢大麻酚(THC)处理组(用10 ng/m L IL-1β预处理同时,添加10-7mol/L雌二醇和10-5mol/L THC),各组处理36 h。实时定量PCR检测滑膜细胞中核因子κB抑制蛋白α(IκBα)和IL-6的mRNA水平,Western blot法检测滑膜细胞中IκBα和IL-6的蛋白水平。结果骨关节炎滑膜细胞中有ERβ表达;IL-1β组IL-6的mRNA和蛋白水平表达均增高;IκBα蛋白水平降低,而mRNA水平升高,同时磷酸化IκBα蛋白水平升高;与IL-1β组相比,IL-1β联合雌二醇组,IL-6、IκBα、p-IκBα蛋白及mRNA水平下降。用IL-1β、雌二醇、雌激素受体阻断剂三者联合组,则可见IL-6、IκBα、p-IκBα蛋白及mRNA水平较IL-1β处理组无明显变化。结论雌二醇通过结合ERβ抑制NF-κB通路激活从而发挥抗炎作用。
Objective To investigate the mechanism of estrogen＇s anti-inflammatory effects on synovial cells during the pathogenic process of osteoarthritis. Methods We isolated synovicytes from synovium tissues and identified the cells with flow cytometry. Then we detected the expression level of estrogen receptor β （ERβ） in synovicytes with immunofluorescence staining. The synovicytes were divided into control group, group pretreated with 10 ng/mL IL-1β, group pretreated with 10 ng/mL IL-1β and 10-7 mol/L estrogen, group pretreated with 10 ng/mL IL-1β, 10-7 mol/L estrogen and specific antagonist of ERβ, 10-5 mol/L tetrahydrocannabinol （THC）. Thirty-six hours later, we observed the mRNA and protein levels of IKBa, phospho-IkBα （p-IkBα） and IL-6. Results Immunofluorescence staining showed the high expression level of ERβ in synovicytes. In IL-1β treated cells, IL-6 mRNA and protein level, IkBα mRNA and p-IkBα protein levels were elevated compared with the control group, while IkBα protein level declined. In the cells pretreated with IL-1β and estrogen, the mRNA and protein levels of IL-6, IkBα and p-kBαa were inhibited compared with IL-1β treated cells. THC blocked the effects of estrogen on the IL-1β and estrogen treated cells, and the mRNA and protein levels of IL-6, IkBα and p-IKBa had no significant difference compared with IL-1β treated cells. Conclusion The estrogen can restrain the activation of NF-kB pathway in synovicytes via ERβ, thus playing a vital role in anti-inflammation.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2016年第12期1605-1609,1614,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81371926)
