中药喘可治对小鼠T淋巴细胞活化的双向调节作用机理研究

Mechanism Study on Chinese Herbal Chuangkezhi on Dual-Directional Modulation of T- Lymphocyte Activation in Mice

目的研究中药喘可治(Chuankezhi,CKZ)对小鼠T淋巴细胞活化的双向调节作用的机理。方法无菌分离小鼠淋巴结并制备单个淋巴细胞悬液,分别加入巴戟天(Bajitian,BJT)提取物(50μg·mL-1)、淫羊藿(Yinyanghuo,YYH)提取物(100μg·mL-1)和喘可治(50μg·mL-1BJT+100μg·mL-1YYH),预孵育4h后,加入刺激剂,12h后,运用流式细胞术结合双色荧光抗体染色技术检测CD3+T淋巴细胞早期活化标志CD69的表达水平。结果与对照组相比,巴戟天提取物处理组中CD3+T淋巴细胞CD69的表达水平明显升高;与刺激组相比,巴戟天提取物可以协同刺激CD69的表达。淫羊藿提取物对静息状态下的CD+3T淋巴细胞CD69的表达作用不明显,但可以抑制刺激剂引起的CD3+T淋巴细胞CD69的表达。与前两者相比,喘可治则呈现明显的双向免疫调节作用,即上调静息状态的CD+3T淋巴细胞CD69的表达,下调刺激状态的CD3+T淋巴细胞CD69的表达。结论喘可治对T淋巴细胞活化的双向调节作用是巴戟天和淫羊藿共同作用的结果,其中巴戟天具有促进免疫应答的作用,而淫羊藿具有抑制免疫应答的作用。

Objective To study the mechanisms of Chinese herbal drug,chuangkezhi（CKZ）on the dual directional modulation of T-lymphocyte activation in mice.Methods The mouse lymph node was separated in sterilization and the single lymphocyte suspension was prepared.Radix Morindae Officinalis extract（50 μg·mL-1）,Epimedium brevicornum Maxim extract（100 μg·mL-1）and CKZ（50 μg·mL-1 Radix Morindae Officinalis ＋ 100 μg·mL-1 Epimedium brevicornum Maxim）were added separately.After pre-incubation for 4 h,the stimulator was added.12 h later,the flow cytometry in combination with double-color fluorescence antibody staining technology were used to detect the expression of CD69,the early activated marker of CD3＋T lymphocyte.Results Compared with blank control group,in Morindae Officinalis extract treatment group,CD69 expression in CD3＋T lymphocyte was upregulated apparently.Compared with stimulation group,Morindae Officinalis extract co-stimulated CD69 expression.Epimedium brevicornum Maxim extract did not impact apparently CD69 expression in CD3＋T lymphocyte at the resting state,but it inhibited CD69 expression of CD3＋T lymphocyte induced by the stimulator.As compared with the two extracts,CKZ indicated obvious dual immune modulation,meaning that it upregulated CD69 expression of CD3＋T lymphocyte at resting state and downregulated CD69 expression of CD3＋T lymphocyte at stimulating state.Conclusion The dual-directional modulation of CKZ on CD3＋T lymphocytes activation Results from the cooperation of Morindae Officinalis and Epimedium brevicornum Maxim,in which Morindae Officinalis promotes immune response and Epimedium brevicornum Maxim inhibits immune response.