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目的:研究痹肿消汤(Bizhongxiao decoction,BZXD)有效成分之一芹菜素(Apigenin,AP)对小鼠体外活化T细胞活化、增殖及细胞因子基因表达的影响,探讨其免疫抑制作用及治疗类风湿性关节炎(Rheumatoid ar-thritis,RA)方面的可能机制。方法:无菌制备小鼠脾脏及淋巴结T淋巴细胞,加入不同浓度的AP(0、20、40、80、120、160μmol.L-1)并用佛波酯(Phorbol-12-myristate13-acetate,PMA)和离子霉素(Ionomycin,Ion)刺激T细胞活化增殖;48h后用MTT法检测T细胞增殖情况,6 h后用RT-PCR检测白细胞介素-2(Interleukin-2,IL-2)mRNA、干扰素-γ(Interferon gamma,IFN-γ)mRNA及IL-4mRNA的表达。结果:48h后,PMA+Ion组细胞MTT吸光度(Absorbance,A值)明显高于空白对照组(P<0.01)。40、80、120、160μmol.L-1浓度的AP组各吸光度均较PMA+Ion组减弱(P<0.01)。在PMA+Ion刺激6h后,PMA+IOn组细胞IL-2mRNA,IFN-γmRNA及IL-4mRNA表达较空白对照组增强,比较均有显著差异性(均P<0.01)。40、80、120μmol.L-1浓度的AP组的上述各细胞因子基因的表达均较PMA+Ion组明显减弱(均P<0.01),且呈不同程度的抑制,具有浓度相关性。20μmol.L-1浓度的AP组的IFN-γmRNA及IL-4mRNA的表达较PMA+Ion组表达明显减弱(P<0.05),但20μmol.L-1浓度的AP组IL-2mRNA的表达较PMA+Ion组无明显减弱(P>0.05)。结论:AP对T细胞的增殖及IL-2mRNA、IFN-γmRNA及IL-4mRNA表达具有抑制作用,提示AP具有免疫抑制β免疫调节作用,可能为痹肿消汤治疗RA的有效成分之一。
OBJECTIVE: To study the effect of Apigenin (AP), an active ingredient of Bizhongxiao decoction (BZXD), on the activation and proliferation of activated T cells in vitro and the gene expression of cytokines in vitro, and to explore its immunosuppressive effects and therapeutic effects Possible Mechanisms in Rheumatoid Arthritis (RA). Methods: The spleen and lymph node T lymphocytes were prepared aseptically. Different concentrations of AP (0, 20, 40, 80, 120 and 160μmol.L-1) were added and incubated with Phorbol-12-myristate 13- ) And ionomycin (Ion) were used to stimulate the proliferation of T cells. After 48 h, the proliferation of T cells was detected by MTT assay. Interleukin-2 (IL-2) mRNA was detected by RT- , Interferon gamma (IFN-γ) mRNA and IL-4 mRNA. Results: The MTT absorbance (Absorbance, A value) of PMA + Ion group was significantly higher than that of blank control group after 48h (P <0.01). The absorbance of 40, 80, 120 and 160μmol.L-1 groups were weaker than that of PMA + Ion group (P <0.01). After PMA + Ion stimulation for 6h, the expression of IL-2mRNA, IFN-γmRNA and IL-4mRNA in PMA + IOn group were significantly higher than those in blank control group (all P <0.01). Compared with PMA + Ion group, the expressions of these cytokines genes in 40, 80 and 120μmol.L-1 AP groups were significantly decreased (all P <0.01), and were inhibited to a certain extent with a concentration-dependent manner. Compared with PMA + Ion group, the expression of IFN-γmRNA and IL-4mRNA in 20μmol.L-1 AP group was significantly decreased (P <0.05), but the expression of IL-2mRNA in 20μmol.L-1 AP group was lower than PMA + Ion group no significant weakened (P> 0.05). CONCLUSION: AP can inhibit the proliferation of T cells and the expression of IL-2mRNA, IFN-|ÃmRNA and IL-4mRNA, suggesting that AP possesses immunosuppressive β immunoregulation and may be one of the active ingredients of bazhongxiao decoction in the treatment of RA.