细辛脂素体外免疫抑制作用的实验研究

Experimental study of the effects of Asarinin on immunosuppression activity in vitro

目的 探讨细辛体外免疫抑制作用的有效成分和作用机制并与环孢素 (CsA)进行比较。方法 提取中药细辛中的细辛脂素 ,分离成年Wistar大鼠的心肌细胞 (2× 10 6个 /ml)和SD大鼠的脾细胞 (1× 10 7个 /ml ) ,前者做刺激细胞 ,后者做反应细胞 ,各 0 1ml于 96孔板进行混合细胞培养 ,加入 0 1ml含细辛脂素大鼠血清 ,培养 10 8h后MTT法测定脾细胞增殖抑制率 ;培养液上清中IL 2、IFN γ及IL 4水平 ;并用光镜、电镜观察心肌细胞。结果 含细辛脂素的动物血清可在体外抑制脾细胞增殖反应 (抑制率达 6 0 37% ) ,心肌细胞损伤减轻 ,使培养液上清中IL 2和IFN γ浓度[(6 9 11± 17 4 7)pg/ml;(183 11± 95 2 4 ) pg/ml]与阳性对照组相比降低 [(16 0 4 4± 19 79) pg/ml;(5 2 1 89± 133 18) pg/ml],IL 4浓度 [(5 3 14± 11 80 ) pg/ml]与阳性对照组相比 [(14 4 4± 4 2 1)pg/ml]升高。与CsA组相比各指标相似 [IL 2 :(5 3 11± 18 35 )pg/ml;IFN γ :(16 6 2 2± 6 1 5 6 ) pg/ml;IL 4 :(44 2 0± 11 5 6 ) pg/ml],P >0 0 5。 结论 中药细辛提取物细辛脂素与CsA有相似的免疫抑制作用 ,并能够产生保护受体器官作用 ;细辛提取物体外抗排斥反应时IL 2和IFN γ降低的同时使IL 4升高可能与提高移植耐受有关。

Objective To test the extract of Asarum heterotropides called Asarinin in the immunosuppression activity in vitro and compared with cyclosporine A(CsA). Methods Cardiomyocytes of adult wistar rats and spleen cells of adult SD rats were separated. 0.1 ml of the cardiomyocytes(2×10 6/ml) as stimulate cells and 0.1 ml of spleen cells(1×10 7/ml) as response cells were cultured in 96 culture plates. The model of rejection reaction in vitro was set up. Then the 0.1 ml serum including Asarinin was given to culture plates, the inhibition rate of lymphocyte transformation was investigated with MTT after 108 hours and IL-2、INF-γ、IL-4 were detected with ELISA. Results Asarinin inhibited the lymphocyte transformation (inhibition rate 60.37%) in vitro. Compared with positive control group, it significantly decreased IL-2(69.11±17.47 pg/ml vs 160.44±19.79 pg/ ml, P <0.01)、INF-γ(183.11± 95.24 pg/ml vs 521.89± 133.18 pg/ml, P <0.01) and increased IL-4(53.14±11.80 pg/ml vs 14.44± 4.21 pg/ml, P <0.01)in culture plates. The effect was similar to CsA group (IL-2: 53.11±18.35 pg/ml, IFN-γ: 166.22± 61.56 pg/ml, IL-4: 44.20±11.56 pg/ml, P >0.05). Conclusion The extract of Haerba Asari has immunosuppression activity and can inhibit acute rejection in vitro. At the same time it decreases IL-2、 INF-γ and increases IL-4 in culture plates. This may be related to the increase of transplantation tolerance.