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目的:观察四物汤等6个方剂不同提取物对小鼠离体脾淋巴细胞转化增殖能力的影响,探讨小鼠脾淋巴细胞模型在中药复方免疫活性成分筛选中的应用。方法:制备四物汤、四君子汤、六味地黄汤、玉屏风散、桂枝汤、龙胆泻肝汤等6个方剂的水提物、水提醇沉物、醇提物,选择其没有明显细胞毒性的浓度范围,在96孔板中分别与小鼠正常的和经PMW诱导的小鼠脾淋巴细胞共同孵育,以MTT法检测细胞转化增殖情况。结果:四物汤上述三种提取物、四君子汤与六味地黄汤水提物和水提醇沉物、龙胆泻肝汤水提物能明显增强经PMW诱导的小鼠脾淋巴细胞转化增殖能力(P<0.05或P<0.01);桂枝汤醇提物能明显抑制经PMW诱导的小鼠脾淋巴细胞转化增殖能力(P<0.01);仅有四物汤水提物能明显增强小鼠正常脾淋巴细胞转化增殖能力(P<0.01),其活性明显较PMW诱导组低(P<0.01)。上述方剂刺激淋巴细胞转化增殖的效应具有提取物和浓度的差异性。结论:在中药复方免疫活性物质的筛选中,丝裂原刺激下的小鼠脾淋巴细胞模型作用稳定,应用时注意选择没有明显细胞毒性的药物浓度。
OBJECTIVE: To observe the effects of different extracts of Siwu Tang and other six prescriptions on the transformation and proliferation of splenic lymphocytes in mice, and to explore the application of mouse splenic lymphocyte model in the screening of immunocompetent components of traditional Chinese medicine. Methods: The water extracts of six prescriptions including Siwu Tang, Sijunzi Decoction, Liuwei Dihuang Decoction, Yupingfeng Powder, Guizhi Decoction and Longdan Xiegan Decoction were prepared. Cytotoxicity concentration range, respectively, in normal 96-well plates and mice were induced by PMW spleen lymphocytes were incubated with MTT assay cell proliferation and proliferation. Results: The three extracts of Siwu Tang, Sijunzi Decoction and Liuweidihuang Decoction water extract and water extract alcohol precipitation, Long Dan Xie Gan Decoction water extract can significantly enhance the ability of PMW-induced mouse spleen lymphocyte transformation and proliferation (P <0.05 or P <0.01). The ethanol extract of Cassia twig could significantly inhibit the proliferation and proliferation of splenic lymphocytes induced by PMW (P <0.01). Only the water extract of Siwu decoction could significantly enhance the normal splenic lymph (P <0.01). Its activity was significantly lower than that of PMW induction group (P <0.01). The effect of the above prescriptions on the proliferation of lymphocytes to stimulate proliferation has differences in extract and concentration. Conclusion: In the screening of immunocompetent substances in Chinese herbal compound, the effect of mitogen-stimulated mouse splenic lymphocyte model is stable, and in the application, the concentration of drug without obvious cytotoxicity is selected.