雌激素抑制淋巴细胞IL-1β和IL-6释放并下调Cx40及Cx43水平

Estrogen inhibits the release of IL-1β and IL-6 and down-regulates the expression of Cx40 and Cx43 on lymphocytes

目的 探讨雌激素(E)对淋巴细胞上连接蛋白40(Cx40)和Cx43表达及白细胞介素1β(IL-1β)和IL-6释放的影响。方法 选取Wistar京都(WKY)大鼠6只,采集腹主动脉外周血,分离淋巴细胞进行培养,分为空白组(Ctrl)、刀豆蛋白A(ConA)组和刀豆蛋白A+雌激素(ConA+E)组。应用ELISA检测培养基上清液中IL-1β、IL-6的水平;应用免疫荧光技术检测淋巴细胞上Cx40和Cx43的表达和分布情况;应用流式细胞术检测淋巴细胞上Cx40和Cx43表达水平变化;应用Western blot检测淋巴细胞上Cx40和Cx43的蛋白表达。结果 ConA组IL-1β和IL-6表达较Ctrl组升高(P<0.01),ConA+E组较ConA组降低(P<0.01,P<0.05);免疫荧光结果显示Cx40主要表达在细胞质及细胞核,而Cx43主要表达在细胞膜,ConA组淋巴细胞上Cx40和Cx43蛋白表达较Ctrl组升高(P<0.01),ConA+E组较ConA组降低(P<0.01);ConA组淋巴细胞上Cx40和Cx43表达水平较Ctrl组升高(P<0.01,P<0.05),ConA+E组较ConA组降低(P<0.05);ConA组淋巴细胞上Cx40和Cx43蛋白表达较Ctrl组明显升高(P<0.01),ConA+E组较ConA组明显降低(P<0.01)。结论 E抑制炎症因子IL-1β和IL-6的释放,同时下调淋巴细胞上Cx40和Cx43的表达,提示Cx40和Cx43可能参与E的抑炎作用。

Objective To investigate the effects of estrogen(E)on the expression of connexin 40(Cx40)and Cx43 on lymphocytes and the release of interleukin 1β(IL-1β)and interleukin 6(IL-6).Methods Peripheral blood lymphocytes of the WKY rats were isolated and cultured from abdominal aorta,then divided into Control group,ConA group,and ConA+E group.The level of IL-1βand IL-6 were detected by ELISA.Immunofluorescence technique was used to detect the expression and location of Cx40 and Cx43 on lymphocytes;flow cytometry was used to detect the expression of Cx40 and Cx43 on lymphocytes;expression of Cx40 and Cx43 on lymphocytes was detected by Western blot.Results The level of IL-1βand IL-6 was higher in ConA group than Control group(P<0.01),lower in ConA+E group than ConA group(P<0.01,P<0.05).Immunofluorescence results showed that Cx40 was mainly expressed in cytoplasm and nucleus of lymphocytes,while Cx43 was mainly expressed on the membrane of lymphocytes.The expression of Cx40 and Cx43 on lymphocytes was significantly higher in ConA group than Control group(P<0.01),and lower in ConA+E group than ConA group(P<0.01).The expression of Cx40 and Cx43 on lymphocytes was higher in ConA group than Control group(P<0.01,P<0.05),and lower in ConA+E group than ConA group(P<0.05).The protein expression of Cx40 and Cx43 on lymphocytes was significantly higher in ConA group than Control group(P<0.01),significantly lower in ConA+E group than ConA group(P<0.01).Conclusion Estrogen inhibits the release of IL-1βand IL-6 from lymphocytes and down-regulates the expression of Cx40 and Cx43 on lymphocytes,suggesting that Cx40 and Cx43 on lymphocytes may be involved in the anti-inflammatory effect of E.