脱氢表雄酮对鼠成骨细胞及CD4+T细胞协同刺激分子表达的影响

目的探讨脱氢表雄酮（DHEA）对成骨细胞（osteoblasts，OB）及CD4^＋T细胞表达协同刺激分子的调控作用。方法颅骨酶解法培养鼠OB，体外模拟雌激素撤退；免疫磁珠细胞分选（rnagnetic cell soaing，MACS）法分离CD4^＋T细胞；将OB或CD4^＋T细胞分为对照组、E2及DHEA处理组，并以LPS刺激；以流式细胞术分析OB表面CD80、CD86以及CD4^＋T细胞表面CD28、CTLA-4的表达。结果经E2及DHEA处理后，OB表达CD80、CD86显著增加（P〈0．05，P〈0．01）；CSA可降低对照组及DHEA处理组OBCDS0、CD86的表达（P〈0．01）。除DHEA组CD28^＋T细胞百分比增加外（P〈0．05），其余各组CD4^＋T细胞CD28和CTLA-4的表达无显著改变（P〉0．05）。结论DHEA可上调鼠OB协同刺激分子CD80、CD86表达，该作用可被CsA阻滞；DHEA还上调CD4^＋T细胞CD28的表达，提示可改善骨．免疫调节网络。

Effect in vitro of dehydroepiandrosterone on the costimulatory molecule expression of murine osteoblasts and CD4 + T lymphocytes

Objective To investigate modulation of DHEA in the costimulatory molecule of the osteoblasts(OB) and CD4~+ T lymphocytes. Methods The OBs were isolated from calvaria of neonatal BALB/c mice, and cultured by the enzyme-digested method. The CD4~+ T cells were isolated from spleen with MACS separation. The OBs or CD4~+ T cells were treated with E2 or DHEA and then stimulated by LPS. The expression of costimulatory molecules on the OBs and CD4~+ T lymphocyte were examined by flow cytometry(FCM). Results The expression of the costimulatory molecule CD80, CD86 on the OBs significantly increased after treatment with E2 or DHEA(P<0.05 or P<0.01). The expression of the costimulatory molecules like CD80, CD86 on OBs in the DHEA-treating group and in the control were suppressed significantly by CsA in vitro(P<0.01). DHEA significantly increased the expansion of the CD4~+CD28~+ T cells(P<0.05). Conclusion DHEA up-regulates the expression of costimulatory molecule CD80 and CD86 on OBs, that can be blocked by CsA. DHEA also increases the expansion of the CD4~+CD28~+ cells, and likely improves the osteo-immune network.