左归丸逆转Th1/Th2亚群偏移减轻雌激素缺乏骨丢失

Mechanism of Zuogui pills on the shift of Th1/Th2 to set-back bone loss caused by estrogen deficiency

目的探讨雌激素缺乏状态下Th1/Th2亚群偏移与骨丢失的相关性及左归丸逆转Th1/Th2亚群偏移减轻雌激素缺乏骨丢失的作用机制。方法纳入未绝经妇女、绝经妇女、绝经中药组妇女各15例,电化学发光法检测血清E2水平;分离静脉血单个核细胞,流式检测Th1/Th2亚群比例;Western blot检测Th1/Th2亚群特异性核转录因子(T-bet、GATA-3)蛋白表达;RT-PCR法检测血清Th1/Th2亚群特征性细胞因子(TNF-α、IL-4)mRNA表达水平;双能X线骨密度仪测定研究对象腰椎2～4(L2～4)前后位BMD。结果与未绝经妇女比较,绝经妇女血清E2表达及BMD均明显降低(P<0.05),Th1亚群比例明显升高(P<0.05),Th2亚群比例明显降低(P<0.05),T细胞向Th1亚群偏移。相关性分析显示,BMD与E2水平、Th2亚群比例呈显著正相关(P<0.05),与Th1亚群比例呈显著负相关(P<0.05);E2水平与Th1亚群比例呈显著负相关(P<0.05),与Th2亚群比例呈显著正相关(P<0.05)。与绝经组比较,左归丸治疗后绝经妇女腰椎BMD明显升高(P<0.05),Th1亚群比例明显降低(P<0.05),Th2亚群比例明显升高(P<0.05),T细胞向Th2亚群偏移,T-bet蛋白表达明显降低(P<0.05),GATA-3蛋白表达明显升高(P<0.05),TNF-αmRNA表达降低(P<0.05),IL-4mRNA表达升高(P<0.05),血清E2表达无显著性差异(P>0.05)。结论雌激素缺乏状态下Th1/Th2亚群偏移与骨丢失密切相关,左归丸调节T细胞亚群特异性核转录因子表达,逆转雌激素缺乏导致的Th1亚群偏移状态,减少骨丢失,促进骨形成。

Objective To analyse the correlation between shift of Th1 / Th2 and bone loss in the state of estrogen deficiency and the mechanism of Zuogui pills set-backing the shift of Th1 / Th2 to alleviate bone loss caused by estrogen deficiency.Methods 15 cases of premenopause women,15 cases of postmenopause women and 15 cases of postmenopause women taking Zuogui pills were selected in this study.The serum estradiol（E 2） were detected by electrothemiluminescence immunoassay.Venous blood mononuclear cells were isolated by Ficoll density gradient centrifugation,and the percentages of Th1 / Th2 subsets were detected by flow cytometry.T-bet and GATA-3 were detected by Western blot.TNF-αmRNA and IL-4 mRNA were detected by RT-PCR.BMD of lumbar vertebrae2～4 was measured with dual energy X ray absorptiometry.Results Compared with premenopause women,the expression of E 2 and BMD of postmenopause women were decreased obviously（P0.05）.The percentage of Th1 subset was increased obviously（P0.05）,and the percentage of Th2 subset were decreased obviously（P0.05）.T-cells subsets offset into Th1 subset.BMD were positively correlated with the expression of E2 and the percentage of Th2 subset（P0.05）.BMD were negatively correlated with the percentage of Th1 subset（P0.05）.The expression of E 2 was negatively correlated with the percentage of Th1 subset（P0.05）.The expression of E 2 was positively correlated with the percentage of Th2 subset（P0.05）.After taking Zuogui pills,BMD of postmenopause women were increased obviously（P0.05）.The percentage of Th1 subset were decreased obviously（P0.05）.And the percentage of Th2 were increased obviously（P0.05）.T-cells subsets offset into Th2 subset.The expression of T-bet were decreased obviously（P0.05）.The expression of GATA-3 were increased obviously（P0.05）,and the mRNA expression of TNF-αwere decreased obviously（P0.05）.The mRNA expression of IL-4 was increased obviously（P0.05）.There was no significant difference in the expression of serum E 2（P0.05）.Conclusion The bone loss is closely related to Th1 / Th2 subsets shift in the state of estrogen deficiency.Zuogui pills regulates cytokines expression in T-cell subset,reverses the shift of Th1 subset due to estrogen deficiency,and inhibits the loss and formation of bone.