苦参碱抑制JM细胞株增殖和诱导凋亡的研究

目的 :观察苦参碱对T细胞白血病细胞株JM的作用。方法 :Wright Giemsa染色及Hoechst 332 5 8荧光染色观察苦参碱作用JM细胞株前后形态学变化 ,电镜观察超微结构的改变 ;流式细胞仪分析不同剂量加药组第4天及 0.8g·L^-1加药组 1～4d细胞周期的改变 ;DNA琼脂糖凝胶电泳检测“梯状”DNA。结果 :第 3天起各加药组细胞形态学观察见典型凋亡特征改变 ,随时间延长更加明显 ;流式细胞仪检测第 4天各组均可见亚二倍体峰。0.1,0.2,0.4,0.6,0.8g·L^-1各处理组凋亡细胞比率分别为 3.1% ,2.5 % ,13.3% ,40.4 % ,48.6 % ,对照组为1.4 % ;各处理组S期细胞比率分别为 28.9% ,26.1% ,27.7% ,20.9% ,14.2% ,对照组为 30.4% ;各处理组G1期细胞比率分别为 63.2% ,67.5% ,68.1% ,75.2% ,83.6% ,对照组为 41.8%。 0 .8g·L^-1加药组 1～4d ,G1期细胞比率分别为 45.5% ,77.3% ,77.2% ,83.6% ;S期细胞比率分别为 28.6 % ,17.5 % ,19.1% ,14.2% ;凋亡细胞比率分别为 3.0% ,3.7% ,9.1% ,48.6%。DNA电泳 :0.4,0 .6 ,0.8g·L^-1组见DNA“梯形”图谱 ,0.1,0.2g·L^-1及对照组未见。结论 :苦参碱可以抑制JM细胞的DNA合成 ,造成G1期阻滞 ,从而抑制了细胞的增殖。同时 ,该生物碱还可诱导。

Matrine Effects on JM Cells by Inhibiting Proliferation and Inducing Apoptosis

Objective: To study effects of matrine on JM cell strain. Method: Morphologic changes were observed under light microscope with Wright Giemsa staining, fluorescence microscope with Hoechst 33258 staining and electron microscope. Alteration of cell cycle of different dose treating groups at the fourth day and 0.8mg·mL^-1 treatment group at the first, second, third, fourth day was analyzed by Flow cytometry. DNA ladder was detected with gel electrophoresis. Result: From the third day after treatment of matrine, typical apoptosis features of cells were observed under light microscope and electron microscope in all test groups, and the features were more prominent with the time prolonging. At fourth day, flow cytometry analysis showed that there were sub G1 peaks in all groups. From 0.1, 0.2, 0.4, 0.6 to 0.8 g·L^-1 treatment groups , the rate of apoptotic cells to total cells were 3.1%, 2.5%, 13.3%, 40.4%, 48.6%, respectively, and what in the control group was 1.4%;the rate of S phase cells to total cells was 28.9%, 26.1%, 27.7%, 0.9%, 14.2%,what in the control group was 30.4%;the rate of G1 phase cells to total cells was 63.2%, 67.5%, 68.1%, 75.2%, 83.6%,what in the control group was 41.8%;From the first, second, third to fourth day, the rate of apoptotic cells to total cells of 0.8 mg·mL^-1 treatment group were 3.0%, 3.7%, 9.1%, 48.6%, respectively; the rate of S phase cells to total cells was 28.6%, 17.5%, 19.1%, 14.2%; the rate of G1 phase cells to total cells were 45.5%, 77.3%, 77.2%, 83.6%. Gel electrophoresis displayed "DNA ladder" in 0.4, 0.6, 0.8 g·L ^-1 groups, while 0.1 and 0.2 g·L ^-1 groups didn't show such result. Conclusion: Matrine can repress DNA synthesis and arrest JM cell strain at G1 phase, sequentially inhibiting the proliferation of the cell. Besides, this alkaloid can induce the apoptosis of JM cells.