积雪草免疫抑制活性成分的分离纯化及效应研究

摘要

AIM:To isolate and purify the active components from Centella asiatica, and to observe their effect on the proliferation and apoptosis of mouse spleen lymphocytes .METHODS:Semi-preparative high-performance liquid chro-matography was performed to further isolate and purify the extracts from Centella asiatica.The effects of the active components on apoptosis and mitochondrial membrane potential of the spleen lymphocytes were assessed by flow cytometry .The structures of the active components were identified by ultraviolet spectrometry , electrospray ionization negative-ion mass spectrometry , [1H] nuclear magnetic resonance (NMR) and [13C] NMR.RESULTS:The molecular weights of the active components from Centella asiatica, named compound I and compound II , were 302 and 286, respectively.The compound I and com-pound II from Centella asiatica significantly inhibited the proliferation , promoted the apoptosis and reduced the mitochondri-al membrane potential of mouse spleen lymphocytes .Compound I and compound II were identified as quercetin and kaempferol .CONCLUSION:The compound I and compound II from Centella asiatica show antiproliferative and immuno-suppressive effects on mouse spleen lymphocytes by promoting apoptosis and reducing mitochondrial membrane potential .%目的：对积雪草的免疫抑制活性成分进行分离、纯化及鉴定，并观察其对小鼠脾淋巴细胞增殖和凋亡的影响，探讨其免疫调节机制。方法：利用半制备型液相色谱对积雪草提取物进一步分离和纯化。运用流式细胞术观察积雪草活性成分对脾淋巴细胞凋亡和线粒体膜电位的影响。利用紫外光谱、电喷雾电离负离子模式质谱、[1 H ]核磁共振（ NMR）、[13 C] NMR等数据鉴定所得活性成分结构。结果：（1）所得积雪草活性成分化合物Ⅰ、Ⅱ的分子量分别为302和286。（2）积雪草中化合物Ⅰ、Ⅱ能显著抑制小鼠脾淋巴细胞增殖。（3）积雪草中化合物Ⅰ、Ⅱ能促进小鼠脾淋巴细胞凋亡，降低线粒体膜电位。（4）[1H] NMR和[13C] NMR鉴定化合物Ⅰ、Ⅱ分别为槲皮素和山奈酚。结论：从积雪草中提取获得的黄酮化合物Ⅰ、Ⅱ分别为槲皮素及山奈酚，具有抑制小鼠脾淋巴细胞增殖的效应，其作用途径可能是促进脾淋巴细胞凋亡并降低线粒体膜电位。