异甘草素诱导人黑色素瘤A375细胞凋亡研究

异甘草素诱导人黑色素瘤A375细胞凋亡研究
阎新燕;司玲玲;高彩霞;于丽娜;王艳明;郑秋生
【期刊名称】《中国药理学通报》
【年(卷),期】2015(000)010
【摘要】目的：研究异甘草素诱导人黑色素瘤( A375)细胞凋亡及其初步机制。

方法采用硫氰酸盐B ( SRB)法测定异甘草素对 A375细胞生长的抑制效应, AO/EB 和Hoechst 33258荧光染色法观察细胞凋亡形态,流式细胞技术检测A375细胞凋亡率；荧光染料二乙酸-2’,7’-二氯荧光素(DCFH-DA)测定A375细胞内活性氧的变化。

5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯并咪唑羰花青碘化物(JC-1)测定线粒体膜电势的变化,利用试剂盒检测异甘草素处理后的A375细胞内ATP和培养液中的乳酸、葡萄糖含量。

结果异甘草素能抑制A375细胞的增殖,并呈浓度依赖性；在荧光显微镜下发现,异甘草素处理后的A375细胞出现明显的凋亡形态；细胞凋亡率随异甘草素浓度的增加而升高；异甘草素处理后的细胞内活性氧有明显上升趋势,细胞线粒体膜电位明显下降；异甘草素作用后胞内ATP含量下降( P<0．05或
P<0．01),培养液中的乳酸含量下降,葡萄糖含量上升(P <0．05或P<0．01)。

结论异甘草素能够抑制A375细胞的恶性增殖,最终诱导细胞凋亡。

推测异甘草素通过活性氧升高,引起线粒体膜电势下降,从而影响A375细胞的糖酵解途径而导致细胞凋亡。

%Aim To evaluate the mechanism of apopto-sis induced by the isoliquiritigenin in A375 human ma-lignant melanoma cells. Methods Sulforhodamine B ( SRB) method was used to determine the A375 cell viability;acridine orange/ethidium bromide ( AO/EB) and Hoechst 33258 staining were used to observe the morphological changes of apoptotic
cells; flow cytome-try was used to detect A375 cell apoptotic rate;DCFH-DA was applied to determine the changes of total intra-cellular ROS in
A375 cells;JC-1 method was used to measure the changes of mitochondrial membrane poten-tial;the kits methods were used to determine the con-tent of ATP, lactic acid and glucose in A375 cell which was treated with different concentrations of isoliquiritigenin. Results Isoliquiritigenin could in-hibit A375 cell proliferation in a concentration-depend-ent manner;
A375 cells showed obvious apoptosis&nbsp;charateristics after treatment by isoliquiritigenin, and the apoptosis rate increased with increasing concentra-tion of isoliquiritigenin. The level of total intracellular ROS in
A375 cells increased obviously after dealing with different concentrations of isoliquiritigenin;in ad-dition, the mitochondrial membrane potential, the lev-els of intracellular ATP,lactic acid and the level of glu-cose uptake all declined. Conclusions These find-ings demonstrate that isoliquiritigenin can induce apop-tosis of A375 cells. The mechanism may be related to elevation of ROS level and reduction of aerobic glycoly-sis level.
【总页数】7页(P1426-1432)
【作者】阎新燕;司玲玲;高彩霞;于丽娜;王艳明;郑秋生
【作者单位】石河子大学药学院，新疆特种植物药资源教育部重点实验室，新疆石河子 832002;石河子大学药学院，新疆特种植物药资源教育部重点实验室，新疆石河子 832002;滨州医学院，山东烟台 264005;滨州医学院，山东烟台264005;石河子大学药学院，新疆特种植物药资源教育部重点实验室，新疆石河
子 832002;石河子大学药学院，新疆特种植物药资源教育部重点实验室，新疆石河子 832002; 滨州医学院，山东烟台 264005
【正文语种】中文
【中图分类】R284.1;R329.25;R739.502.2
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