苦参有效成分对活化慢性湿疹患者Th1细胞Fas-L、IFN-γ表达的影响

Effects of Effective Components in Kuhseng on the Expressions of Fas-L and IFN-γ of Th1 Cells of Chronic Eczema Patients

目的研究中药苦参有效成分(氧化苦参碱和苦参碱)对活化慢性湿疹患者Th1细胞表达Fas配体及IFN-γ表达的影响。方法选取30例慢性湿疹患者为试验组,健康志愿者30名为对照组。两组均从外周血中磁珠分选并纯化Th1细胞,在体外使用免疫刺激剂对细胞进行激活。通过MTT方法筛选出氧化苦参碱和苦参碱对活化的Th1细胞的最大安全浓度,以最大安全浓度稀释5倍,分为氧化苦参碱A、B、C、D、E组和苦参碱A、B、C、D、E组,每组重复3孔,以各浓度中药有效成分干预激活后的Th1细胞,然后用ELISA检测干预前后细胞培养液上清中IFN-γ和Fas-L的含量变化。结果与对照组比较,试验组经免疫刺激剂激活后,IFN-γ浓度显著升高(P<0.01)。苦参碱及氧化苦参碱各组干预前后比较,苦参碱A组(10 μg/mL)的IFN-γ浓度显著升高(P<0.05),氧化苦参碱A、B、C、D、E组的IFN-γ浓度差异无统计学意义(P>0.05);使用≥0.625 μg/mL苦参碱(苦参碱A、B、C、D、E组)的Fas-L浓度显著降低(P<0.05),使用≥0.025 μg/mL氧化苦参碱(氧化苦参碱A、B、C、D组)的Fas-L浓度显著降低(P<0.05)。结论一定浓度的中药苦参有效成分可下调慢性湿疹外周血中活化的Th1细胞Fas-L的表达水平,起到临床治疗作用。

Objective To study the effects of active ingredients in Kuhseng(oxymatrine and matrine) on the expressions of Fas ligand(Fas-L) and IFN-γ Th1 cells in chronic eczema patients. Methods Thirty chronic eczema patients were recruited as a test group and 30 healthy volunteers recruited as the control group. Th1 cells were isolated from their peripheral blood by magnetic separation and Th1 cells purified. They were activated by in vitro immune stimulating agent. MTT method was used to screen the maximum safety concentrations of oxymatrine and matrine on activated Th1 cells. Then they were diluted 5 times the maximum safety concentrations. Later cells were further divided into oxymatrine A, B, C, D, E groups, and matrine A, B, C, D, E groups. Three pores were repeated in each group. Activated Th1 cells were intervened by effective components of each concentration. And then the concentrations of IFN-gamma and Fas-L in cell culture supernatant were detected by ELISA. Results Compared with the control group, the concentration of IFN-γ increased significantly after activated by immune stimulating agent in all test groups(P<0.05). When compared between before and after intervention in oxymatrine and matrine intervention groups, the concentration of IFN-gamma significantly increased in matrine intervened group A(10 μg/mL, P<0.05); no significant difference was shown among matrine intervention groups A, B, C, D, and E(P>0.05). Fas-L concentration was significantly reduced in matrine intervention groups A, B, C, D and E(the concentration of matrine equal to or more than 0.625 μg/mL). Fas-L concentration was significantly reduced in oxymatrine intervention groups A, B, C and D(the concentration of matrine equal to or more than 0.025 μg/mL, P<0.05). Conclusion The effective components at a certain concertration of Kuhseng played a role in clinical treatment by down-regulating the expression levels of Fas-L in activated Th1 cells of peripheral blood in chronic eczema patients.