摘要
目的:探讨脱氢表雄酮(DHEA)对去势鼠CD4+T细胞Th1/Th2型细胞因子表达和成骨细胞(OB)增殖的影响。方法:建立小鼠绝经后骨质疏松(PMO)模型,分为OVX组、OVX+DHEA组、OVX+E2组。另设Sham组为假手术对照。取腰椎和股骨行骨组织形态计量分析;流式细胞术(FCM)检测各组鼠脾脏CD4+T细胞内IL-2、IFN-γ(Th1型)和IL-4、IL-10(Th2型)的表达;并取椎骨植块法培养OB,FCM分析增殖细胞核抗原(PCNA)表达。结果:OVX组与Sham组相比,椎骨和股骨骨小梁面积显著下降(P<0.01),说明PMO模型成功建立;与OVX组相比,OVX+DHEA组腰椎、股骨及OVX+E2组腰椎、股骨骨小梁面积都明显增加(P<0.01)。OVX组IL-2及IFN-γ表达显著低于Sham组(P<0.05);而OVX+DHEA组和OVX+E2组显著高于OVX组(分别为P<0.01和P<0.05)。OVX+DHEA组IL-4及IL-10表达显著低于OVX组(P<0.01)。OVX组IFN-γ/IL-4比值明显低于Sham组(P<0.05);OVX+DHEA组和OVX+E2组IFN-γ/IL-4比值均显著高于OVX组(P<0.01)。OVX组OB的PCNA表达与Sham组相比显著增高(P<0.05);与OVX组相比,OVX+DHEA组OB核内PCNA的平均表达强度和阳性细胞百分率皆显著增高(分别为P<0.05和P<0.01)。OVX+DHEA组OBPCNA表达与CD4+T细胞IFN-γ水平呈明显正相关(r=0.931,P<0.05);与IL-4水平呈负相关(r=-0.815,P<0.05)。结论:DHEA可通过细胞免疫调节作用形成Th1型免疫偏移,从而显著改善PMO的免疫状态;并有效促进OB增殖,显著改善骨形态。
Objectlve：To investigate modulation of DHEA on the expression of Th0/Th2 cytokines in CD4^＋T lymphocytes and the proliferative ability of osteoblasts（OBs） in ovariectomized mice. nethods-BALB/c mice were ovariectomized for foundation of osteoporosis, and then were afforded with dehydroepiandrosterone （OVX ＋ DHEA group）, 17β-estradiol （OVX ＋ E2 group） and Saline （OVX group）, respectively. The femur, vertebra and spleen of each group were collected in 12 weeks of treatment. Femur and vertebra were measured for morphometry of bone tissue. The expression of Thl-type（ IL-2 and IFN-γ） and Th2-type cytokines（ IL-4 and IL- 10） of splenic CD4^＋T cells were analyzed by FCM. The OBs were isolated from vertebra and cultured. The proliferative cell nuclear antigen（PCNA） of OBs was analyzed by flow cytometry（FCM）. Results：The area of bone trabecula in OVX group decreased significantly as compared to that of the Sham group （ P 〈 0. 01 ） ; area of bone trabecula in OVX ＋ DHEA group and OVX ＋ E2 group is higher than that of OVX group（P 〈0. 01 ）. The IL-2 and IFN-γ expression in OVX group decreased remarkably as compared to that of the Sham group（P 〈0. 05）. The Thl cytokine expression in OVX ＋ DHEA group and OVX ＋ E2 group increased significantly as compared to that of the OVX group（ P 〈 0. 01, P 〈 0. 05 ）. The IL-4 and IL-10 expression in splenic CD4^＋ T cells of OVX ＋ DHEA group reduced significantly as compared to that of the OVX group（P 〈0. 01 ）. The ratio of IFN-γ/IL-4 was significantly decreased in the OVX group compared to the Sham group（P 〈0. 05）. Compared to the OVX group, the ratio of IFN--γ/IL-4 was significantly increased in OVX ＋ DHEA group and OVX ＋ E2 group（P 〈0. 01 ）. The expression of PCNA in the OBs of OVX group increased significantly compared to the Sham group（P 〈0. 05）. Compared to the OVX group, the average fluorescence intensity and positive cell percentage of PCNA in the OBs of OVX ＋ DHEA group increased significantly （ P 〈 0. 05 or P 〈 0. 01 ）. In OVX ＋ DHEA group, the expression of PCNA in the OBs was positively related to that of IFN-γ in the CD^＋T cells（r =0. 931 ,P 〈0. 05）, and negatively related to that of IL- 4 （r = -0. 815,P 〈0. 05 ）. Conclusion： DHEA treatment could increase the proliferation of OBs in vivo, improve the bone tissue morphometry of the PMO model. Moreover, DHEA can promote a shift in Th1/Th2 ratio balance toward the Thl-bias, thus improve the immune state of the PMO model.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2007年第6期545-550,共6页
Chinese Journal of Immunology
基金
国家自然科学基金项目(No.30472259)
上海市科技攻关项目(No.004019061)
上海市卫生局青年基金(No.044Y06)资助
