芹菜素对小鼠T细胞增殖及细胞因子基因表达的影响

Effects of Apigenin on the Proliferation and Cytokine Gene Expression of Murine T Lymphocytes

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 effects of AP, which is one of effective constituents of bizhongxiao decoction （BZXD） on the activation, proliferation and cytokine murine T lymphocytes. To explore the mechanisms of it on immanosuppression and the treatment of rheumatoid arthritis （ RA ）. Methods ：Murine T lymphocytes of spleen and lymph nodes were prepared with aseptieism. The lymphocytes were pretreated with AP at different concentrations （0,20,40,80,120, 160 umol · L^-1 ） and stimulated for activation and proliferation with PMA plus Ion. T - cell proliferation was detected by MTT after 48h and the expression of IL-2mRNA ,IFN- γmRNA and IL-4 mRNA were detected by RT- PCR after 6 h. Results ： After 48 hours ,the absorbance of PMA plus Ion group was higher significantly than the blank control group （ P 〈 0.01 ）. The absorbanee of groups of 40,80,120.160umol·L^-1 AP was lower significantly than the PMA plus Ion group （P〈0.01）. After 6 hours,the expression of IL- 2 mRNA, IFN-γ mRNA and IL- 4mRNA of PMA plus Ion group was higher than the blank control troup, and there were significant differences （ all P 〈 0.01 ）. The expression of the above mentioned eytokine gene expression of the groups of 40,80,120umol·L^-1 AP was all significantly weaker than the PMA plus Ion group（ all P 〈0.01 ） and was inhibited at different degree in concentration dependent. The expression of IFN - 3,mRNA and IL -4mRNA on 20umol·L^-1 AP group was weaker significantly than the PMA plus Ion group （P〈 0.05 ） , but there was no signifieandy weak on the expression of IL - 2mRNA （ P 〉 0.05 ）. Conchzsions ： AP had an obvious inhibition on the proliferation and the expression of IL -2mRNA, IFN -γ mRNA and IL -4mRNA of marine T lymphocytes. It shows that AP has the effeet of immunosuppression and immunological regulation, it is one of the effeetive stituent of BZXD on the treatment of RA.