苦参碱诱导淋巴细胞白血病L1210细胞凋亡的实验研究

Study on matrine-induced apoptosis in L1210 cells of lymphocytic leukemia

目的在体外培养条件下,观察苦参碱对L1210细胞的损伤效应,探讨苦参碱的作用机制,期待为临床白血病治疗提供新的思路。方法取对数期生长的L1210细胞,在细胞培养板每孔接种1×105/孔,培养24 h后,加入终浓度为0.5 g/L、1.0 g/L、2.0 g/L的苦参碱,对照组加入等体积的培养液。收集24 h、48 h和72 h的L1210细胞标本,台盼蓝染色观察直接损伤的作用,TUNNEL法观察诱导细胞凋亡的情况。结果苦参碱浓度越高,作用时间越长,L1210细胞台盼蓝拒染率越低,与对照组相比有显著性差异。高浓度的苦参碱(>0.5 g/L)对L1210细胞细胞凋亡才会有显著的影响(P<0.05),随着时间的延长,较高浓度的苦参碱也会显著影响L1210细胞的凋亡。结论苦参碱有杀伤白血病L1210细胞的作用,其机制可能是诱导细胞凋亡和直接损伤细胞膜。

Objective It is to observe the damaging effect of matrine on L1210 cells planted in vitro,to explore its mechanism and to find some new clues for leukemia therapy.Methods L1210 cells growing in log phase were cultured in cell cuture plate at a dosage of 1×105/hole.After 24 hours＇ cuture,matrine at different concentrations（0.5 g/L,1.0 g/L,2.0 g/L） were added.The control group was added with cuture liquid of the same volume.Exams of L1210 cells at 24h,48h and 72h were collected.Derected damage were observed by trypan blue rejecting dyeing（TRD） and cell apoptoiss were observed by TUNNEL respectively.Results When the countent and the functional time of matrine increased,the rate of TRD decreased respetively.Compared with control group,the difference was signifiant（P0.01）.Only high concentration of matrine（0.5 g/L） could have significant effect on L1210（P0.05）.Lower-high concentration of matrine could have influence the apoptosis of L1210 cells along with functional time increasing.Conclusion Matrine can damage L1210 cells via,whose mechanim may be inducing apoptosis and destroying cell membrane.