雷公藤红素对人多发性骨髓瘤H929细胞凋亡的诱导作用及分子机制研究

Study on Induction of Tripterine on Apoptosis of H929 Human Myeloma Cells and Its Molecular Mechanism

目的:观察雷公藤红素诱导人多发性骨髓瘤H929细胞凋亡的效果,并分析诱导凋亡的分子机制。方法:取培养48 h后处于平台期的H929细胞,按照1.0×10^(9)/L细胞密度接种于6孔细胞培养板中,分为对照组、雷公藤红素组(0.5 mg/L、1.0 mg/L和5.0 mg/L),各浓度雷公藤红素组分别加入对应浓度的雷公藤红素,对照组加入等体积二甲亚砜(DMSO)溶液。比较不同浓度雷公藤红素组H929细胞活力抑制率,比较各组H929细胞凋亡率及H929细胞培养物中P53、剪切型半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)、剪切型聚腺苷二磷酸核糖聚合酶-1(cleaved PARP-1)、bax、bcl-2表达水平。结果:H929细胞活力抑制率随着雷公藤红素处理浓度升高而升高,不同浓度雷公藤红素组H929细胞活力抑制率两两比较,差异均有统计学意义(P<0.05)。与对照组比较,不同浓度雷公藤红素组H929细胞凋亡率均较高(P<0.05)。H929细胞凋亡率随着雷公藤红素处理浓度增加而增加,不同浓度雷公藤红素组H929细胞凋亡率两两比较,差异均有统计学意义(P<0.05)。与对照组比较,不同浓度雷公藤红素组H929细胞培养物中P53、cleaved caspase-3、cleaved PARP-1、bax表达水平均较高(P<0.05),bcl-2表达水平均较低(P<0.05)。H929细胞培养物中P53、cleaved caspase-3、cleaved PARP-1、bax表达水平随着雷公藤红素处理浓度升高而升高,bcl-2表达水平随着雷公藤红素处理浓度升高而降低,不同浓度雷公藤红素组H929细胞培养物中P53、cleaved caspase-3、cleaved PARP-1、bax、bcl-2表达水平两两比较,差异均有统计学意义(P<0.05)。结论:雷公藤红素对人多发性骨髓瘤H929细胞具有活力抑制和凋亡诱导作用,且有一定的剂量效应,这一作用可能与线粒体凋亡途径有关。

Objective:To observe the effect of tripterine inducing apoptosis of H929 human myeloma cells,and analyze the molecular mechanism of inducing apoptosis.Methods:H929 cells that are in the plateau stage after 48 hours of cultivation were taken,and were inoculated into 6-well cell culture plates with 1.0×10^(9)/L cell density,which were divided into the control group,the tripterine group(0.5 mg/L,1.0 mg/L,and 5.0 mg/L).Each concentration of the tripterine group was added with corresponding concentration of tripterine,and the control group was added with equal volume of dimethyl sulfoxide(DMSO)solution.The inhibition rates of H929 cell viability at different concentrations of tripterine groups were compared,and the apoptosis rates of H929 cells in each group and the expression levels of P53,cleaved cysteine aspartic protease-3(cleaved caspase-3),cleaved poly ADP-ribose polymerase-1(cleaved PARP-1),bax,and bcl-2 in H929 cell culture were compared among the groups.Results:The inhibition rates of H929 cell viability were increased with the increase of tripterine treatment concentration,and there were significant differences being found in the comparison of the inhibition rates of H929 cell viability in different concentrations of tripterine groups(P<0.05).The apoptosis rates of H929 cells in different concentrations of tripterine treatment group were higher when compared with that in the control group(P<0.05).The apoptosis rates of H929 cells were increased with the increase of tripterine treatment concentration,and there were significant differences being found in the comparison of the apoptosis rates of H929 cells in different concentrations of tripterine groups(P<0.05).The expression levels of P53,cleaved caspase-3,cleaved PARP-1 and bax in H929 cell cultures treated with different concentrations of tripterine were relatively high when compared with those in the control group(P<0.05),and the expression levels of bcl-2 were relatively low(P<0.05).The expression levels of P53,cleaved caspase-3,cleaved PARP-1,and bax in H929 cell cultures were increased with the increase of tripterine treatment concentrations,and the expression levels of bcl-2 were decreased with the increase of tripterine treatment concentrations.There were significant differences being found in the comparison of expression levels of P53,cleaved caspase-3,cleaved PARP-1,bax,and bcl-2 in H929 cell cultures in different concentrations of tripterine groups(P<0.05).Conclusion:Tripterine can inhibit the viability and induce apoptosis of H929 human myeloma cells,and has a certain dose effect,which may be related to the mitochondrial apoptosis pathway.