摘要
目的研究中药黄连解毒汤(HLJDT)对人多发性骨髓瘤(MM)细胞RPMI8226增生和凋亡的作用。方法以含10%胎牛血清的RPMI1640培养基常规培养RPMI8226细胞,分别以不同浓度的HLJDT作用不同的时间后,应用锥虫蓝拒染法测定细胞活力变化,MTT法检测细胞的增生变化;采用流式细胞术测定细胞周期的变化;应用流式细胞仪检测药物细胞的凋亡现象,荧光显微镜下观察AO/EB染色后的细胞凋亡形态变化;比色法检测半胱氨酸蛋白水解酶3(caspase3)活性变化。结果与对照组相比,200～800ng/ml浓度的HLJDT可明显抑制细胞增生影响(P<0.01),并呈时间和剂量依赖性;使G0/G1期细胞增多,S期细胞减少,并呈剂量依赖性改变;MM细胞凋亡百分率显著增高(P<0.01),免疫荧光显微镜下可见典型的凋亡细胞形态学变化;使caspase3活性增强,且呈剂量和时间依赖性(P<0.01)。结论HLJDT能有效抑制MM细胞RPMI8226增生,促其凋亡,G0/G1期细胞比例增加和S期细胞比例减少可能是原因之一;caspase3活性增强可能参与其凋亡过程,具体机制还有待研究。
Objective To investigate the effects of huang-lian-jie-du-tang （HLJDT）, a traditional Chinese herbal medicine, on the proliferation and apoptosis of human multiple myeloma （MM） RPMI8226 cells. Methods RPMI8226 cells were cultured together with various concentrations of HLJDT, the anti-pro- liferation effects were evaluated by trypan-blue staining and MTL cell cycle and apoptosis were detected by flow cytometry, and cells apotosis morphological changes were observed by fluorescence microscope. Caspase3 activity was evaluated by BD ApoAlert caspase colorimetfic assay Kit. Results Compared with the contrast cells, HLJDT showed significant cytotoxie effects on RPMI8226 cells viability and proliferation at various con- centration from 200 to 800ng/ml in a time and dose-dependent manner （P 〈0.01）, and it also showed significant effect on cell apoptosis rate （P 〈0.01）when cultured with HLJDT at the concentrations for 24 h in a dose-dependent manner. Huorescence microscopy showed typical apoptosis features of cells at the same time. Caspase3 activity had been enhanced significantly in a time dose-dependent manner （P 〈0.01）. Conclusion HLJDT can inbibit proliferation and induce apotosis of human MM RPMI8226 ceils significantly, and enhanced caspase3 activity is involved in the mechanism.
出处
《白血病．淋巴瘤》
CAS
2006年第6期412-415,共4页
Journal of Leukemia & Lymphoma
