补肾宁心方诱导去势鼠Treg扩增并促进成骨细胞PCNA表达  

A Chinese herb medicine BSNXD promotes amplification of Treg and proliferation of osteoblasts in the ovariectomized mice

作　　者：王凌[1] 孙婵[1] 邱学敏[1] 李大金[1,2] 

机构地区：[1]复旦大学上海医学院附属妇产科医院暨妇产科研究所,上海200011 [2]海南医学院附属医院,海口570102

出　　处：《现代免疫学》2011年第3期187-191,共5页Current Immunology

基　　金：国家自然科学基金项目(30801502);国家重大科学研究计划项目(2006CB944007);上海市医学领军人才基金资助项目

摘　　要：为了探讨补肾宁心方(BSNXD)对去势鼠脾脏内CD4+CD25+Foxp3+Treg数目及特异性细胞因子CTLA-4、TGF-β及IL-10表达的调控,以及BSNXD对成骨细胞(OB)增殖的影响。建立小鼠绝经后骨质疏松(PMO)模型,随机分为Sham组、OVX组、OVX+BSNXD组和OVX+E2组。取各组腰椎和股骨行骨组织形态计量分析;应用流式细胞术(FCM)评价各组鼠脾脏内CD4+CD25+Foxp3+Treg数目;用免疫磁珠分离小鼠脾脏CD4+CD25+Treg,分析其特异性细胞因子CTLA-4、TGF-β及IL-10的表达;椎骨植块法培养OB,FCM分析OB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+BSNXD组及OVX+E2组腰椎、股骨骨小梁面积都明显增加(P<0.01)。与Sham组相比,去势鼠脾脏CD4+CD25+Foxp3+Treg数目显著降低(P<0.01);其特异性细胞因子CTLA-4、TGF-β及IL-10表达明显下降(P<0.01);OB PCNA表达显著增加(P<0.05)。灌服BSNXD后,CD4+CD25+Foxp3+Treg数目及其特异性细胞因子CTLA-4、TGF-β及IL-10表达显著升高(P<0.01);OB核内PCNA平均荧光强度和阳性细胞百分率均显著增加(分别为P<0.01和P<0.01)。BSNXD逆转了去势手术后外周CD4+CD25+Foxp3+Treg数目下降趋势;显著升高其特异性细胞因子CTLA-4、TGF-β及IL-10的表达,从而显著改善PMO的免疫状态及骨形态。To investigate effects of a Chinese herb medicine BSNXD on the number of spleenic CD4＋CD25＋Foxp3＋Tregs,its specific cytokine expressions of CTLA-4,TGF-β,IL-10 and the proliferative ability of osteoblasts（OBs） in the ovariectomized mice.BALB/c mice were ovariectomized in order to establish a post-menopausal for osteoporosis model of mice（PMO）,and then these mice were subjected to the administration with BSNXD,17β-estradiol and saline,respectively thus randomly dividing the experimental mice into 4 groups.e.i.sham group,OVX group,OVX＋BSNXD group and OVX＋E2 group.The femur,vertebra and spleen of each group were collected 12 weeks after treatment in which the femur and vertebra were measured with morphometry of bone tissues,the number of the CD4＋CD25＋Foxp3＋Tregs isolated by immune magnetic beads and the specific cytokines CTLA-4,TGF-β,IL-10 in spleen were analyzed by FCM.Osteoblasts were isolated from vertebra and cultured,and the expression of proliferative cell nuclear antigen（PCNA） of OBs was analyzed by FCM.It was demonstrated that,the area of bone trabecula in OVX group was decreased significantly as compared to that of the Sham group（P0.01）.Meanwhile,the area of bone trabecula in BSNXD group and E2 group were higher high than that of OVX group（P0.01）.After ovariectomy,there was significantly lower number of CD4＋CD25＋Foxp3＋Tregs in splenic CD4＋ T cells（P0.01）.Moreover,the expression of the CD4＋CD25＋Foxp3＋Tregs specific cytokines CTLA-4,TGF-β and IL-10 were significantly decreased（P0.01）,and the significantly higher expression of PCNA in OBs was found（P0.05）.After treatment with BSNXD,the number of CD4＋CD25＋Foxp3＋Tregs and the expression of cytolines CTLA-4,TGF-β,IL-10 increased significantly（P0.01）.In addition,the average fluorescence intensity and positive cell percentage of PCNA in the OBs also increased significantly（P0.05 or P0.01）.It is evident that administration of BSNXD in the ovariectomized mice could convert the tendency o

关 键 词：补肾宁心方 绝经后骨质疏松症 CD4+CD25+Foxp3+Treg 成骨细胞 增殖 

分 类 号：R392.12[医药卫生—免疫学]