摘 要 目的: 探讨苦参碱诱导C6胶质瘤细胞凋亡的作用特点和可能的基因作用机制。 方法：MTT法测定不同浓度苦参碱对C6细胞增殖的抑制率，求出IC50值；倒置显微镜及透射电镜观察苦参碱诱导C6细胞凋亡的形态学改变；Annexin/FITC染色流式细胞术检测不同浓度苦参碱诱导C6细胞的凋亡率；实时定量PCR芯片（Realtime PCR Array）检测苦参碱作用后C6细胞凋亡相关基因的差异表达；免疫细胞化学及Western blotting方法检测苦参碱对C6细胞caspase3表达的影响。结果：苦参碱对C6细胞增殖的抑制率随着药物剂量的增加（0.1~1.0 mg/ml）而增加（P<005），其IC50为0.715 mg/ml。倒置显微镜观察显示苦参碱可诱导胶质瘤细胞出现凋亡改变，透射电镜观察显示苦参碱诱导胶质瘤细胞出现凋亡的超微形态改变，流式细胞术检测显示胶质瘤细胞的凋亡率随着药物剂量的增加（0.2~1.0 mg/ml）而增大[(3.56±0.73)%~(27.55±1.92)%](P<0.05)。胶质瘤细胞经苦参碱作用后出现显著表达差异基因共68个，其中57个基因表达明显上调、11个基因表达明显下调，其中有22个基因与诱导凋亡的死亡受体相关，15个基因与线粒体途径有关；ICC及Western blotting检测都显示苦参碱可诱导C6胶质瘤细胞caspase3表达的上调(P<0.05)。 结论：苦参碱能诱导C6胶质瘤细胞的凋亡，其机制与死亡受体途径和线粒体途径的众多基因有关。

Abstract Objective: To explore the apoptosis inducing effect of matrine on C6 glioma cells and the related mechanisms. Methods: MTT assay was used to examine the inhibition of C6 glioma cell line by matrine at various concentrations and the IC50 was calculated. Inverted microscope and TEM (transmission electron microscope) were employed to observe the morphological alterations of C6 glioma cells after exposure to matrine; FCM (Flow cytometry) was used to detect the apoptosis rate of C6 glioma cells; and realtime PCR was used to examine the differential expression of related genes. ICC and Western blotting was used to detect the expression of caspase3. Results: MTT showed that the cell inhibition effect of matrine increased with its concentration（0.11.0 mg/ml）（P<0.05）, with the IC50 being 0.715 mg/ml. Inverted microscope and TEM showed that matrine induced apoptosis of C6 glioma cells. FCM showed that the apoptosis rate increased[(3.56±0.73)%-(27.55±1.92)%] with the increase of matrine concentration（0.2-1.0 mg/ml）（P<0.05）. Realtime PCR showed that matrine induced upregulation of 58 genes and downregulation of 11 genes, and 22 of them were related to apoptosisinducing death receptor, 15 of them to apoptosisinducing mitochondrial pathway. ICC and Western blotting showed that matrine induced upregulation of caspase3 expression（P<0.05）. Conclusion: Matrine can induce C6 glioma cell apoptosis and the mechanism might be related to a number of genes involved in death receptor pathway and mitochondrial pathway.

黑龙江省自然科学基金资助项目（No.D 200739）