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背景:川芎嗪抑制血管内皮生长因子的表达,但对其诱导HL-60白血病细胞增殖是否有抑制效应尚需进一步实验。目的:观察川芎嗪对血管内皮生长因子诱导的白血病细胞HL-60细胞增殖的影响。设计:重复测量观察。单位:武汉科技大学医学院。材料:实验于2007-03/06在武汉科技大学医学院分子生物学实验中心完成。人白血病细胞系HL-60细胞购于上海细胞生物研究所。盐酸川芎嗪注射液为无锡市第七制药有限公司产品,批号为011014,硫酸鱼精蛋白注射液购自上海第一生化药业公司,批号为010302,免疫组化试剂盒购自博士德公司。方法:①取对数生长期人白血病细胞系HL-60细胞,加入100μg/L血管内皮生长因子,分别加入终浓度为1.5,15,150mg/L川芎嗪实验培养基,以未加川芎嗪注射液的细胞为空白对照组,只含20mg/L鱼精蛋白的细胞为阳性对照组,同时设立血管内皮生长因子对照组,细胞培养48h后,采用MTT法检测HL-60细胞的生长抑制率。②川芎嗪影响HL-60细胞血管内皮生长因子蛋白表达实验:用终浓度为1.5,15及150mg/L川芎嗪处理HL-60细胞,24h后采用免疫组织化学法计算血管内皮生长因子蛋白阳性细胞表达率。主要观察指标:①HL-60细胞生长抑制率。②血管内皮生长因子蛋白表达情况。结果:①HL-60细胞生长抑制率:川芎嗪15,150mg/L作用血管内皮生长因子诱导的HL-60细胞吸光度值均低于血管内皮生长因子对照组,差异有显著性意义(P<0.05)。②血管内皮生长因子蛋白表达情况:川芎嗪作用HL-60细胞24h后,血管内皮生长因子蛋白随川芎嗪给药浓度升高表达逐渐下调,呈一定依赖性,各川芎嗪浓度干预HL-60细胞血管内皮生长因子蛋白表达阳性细胞表达率与对照组比较差异均有显著性意义(P<0.01)。结论:川芎嗪可抑制血管内皮生长因子诱导HL-60细胞的增殖,并下调血管内皮生长因子蛋白的表达。
BACKGROUND: Tetramethylpyrazine inhibits the expression of vascular endothelial growth factor, but its inhibitory effect on the proliferation of HL-60 leukemia cells still requires further experiments. Objective: To observe the effect of tetramethylpyrazine on proliferation of HL-60 cells induced by vascular endothelial growth factor. Design: Repeat measurement observations. Unit: Wuhan University of Science and Technology Medical School. MATERIALS: The experiment was performed at the Molecular Biology Experimental Center, School of Medicine, Wuhan University of Science and Technology from March to June 2007. Human leukemia cell line HL-60 cells were purchased from Shanghai Institute of Cell Biology. Ligustrazine Hydrochloride Injection is the product of Wuxi No. 7 Pharmaceutical Co., Ltd., batch number is 011014, protamine sulfate injection is purchased from Shanghai No. 1 Biochemical Pharmaceutical Company, batch No. is 010302, and the immunohistochemistry kit is purchased from Dr. METHODS: 1 Human leukemia cell line HL-60 in logarithmic growth phase was taken and 100 μg/L vascular endothelial growth factor was added. The final concentrations were 1.5, 15 and 150 mg/L tetramethylpyrazine experimental medium, but not ligustrazine injection. The cells were blank control group. The cells containing only 20mg/L protamine were positive control group and the vascular endothelial growth factor control group was established. After 48h of cell culture, the growth inhibition rate of HL-60 cells was detected by MTT assay. 2 Ligustrazine effect on vascular endothelial growth factor protein expression in HL-60 cells: HL-60 cells were treated with Ligustrazine at final concentrations of 1.5, 15 and 150 mg/L. After 24 h, VEGF-positive cells were calculated by immunohistochemical method. Expression rate. MAIN OUTCOME MEASURES: 1 Growth inhibition rate of HL-60 cells. 2 Vascular endothelial growth factor protein expression. RESULTS: 1The growth inhibitory rate of HL-60 cells: Ligustrazine 15,150mg/L The vascular endothelial growth factor-induced HL-60 cell absorbance values were lower than those of the vascular endothelial growth factor control group, and the difference was significant (P<0.05). 2 Expression of vascular endothelial growth factor protein: After 24 hours of chuanxiongzin-induced HL-60 cells, the expression of vascular endothelial growth factor protein was gradually decreased with the increasing concentration of ligustrazine, and it showed a certain dependence. Each concentration of tetramethylpyrazine interfered with HL-60 cells. The expression rate of VEGF-positive cells was significantly different from that of the control group (P<0.01). Conclusion: Ligustrazine can inhibit the proliferation of HL-60 cells induced by vascular endothelial growth factor and down-regulate the expression of vascular endothelial growth factor.