黄癸素诱导小鼠黑色素瘤B16细胞凋亡的研究

Apoptosis of B16 melanoma cells induced by berberine α-hydroxy-β-decanoylethyl sulfonate

目的探讨黄癸素诱导小鼠黑色素瘤B16细胞凋亡的作用。方法应用MTT法检测黄癸素对体外培养的B16细胞增殖的抑制作用,观察量效及时效关系;通过Rhodamine123染色、琼脂糖凝胶电泳、Hoechst33258染色、caspase-3和caspase-8活性检测,观察黄癸素诱导细胞凋亡的作用和机制。结果黄癸素抑制B16细胞的增殖,具有明显的时间依赖性和浓度依赖性,作用24、48、72h的IC50分别为16.59、9.29和6.22mg·mL-1;B16细胞经黄癸素处理后,出现染色质固缩、DNALadder等凋亡表现,Rhodamine123染色荧光降低和caspase-3、caspase-8活性增强,提示黄癸素引起线粒体膜电位降低可能触发caspases级联反应而导致细胞凋亡。结论黄癸素可抑制B16细胞增殖,诱导B16细胞凋亡。

Aim To investigate the mechanism of induction of cell apoptosis of berberine α-hydroxy-β-decanoylethyl sulfonate（HB）.Methods MTT assay was employed to determine the cytotoxicity of HB in B16 cells.Through Rhodamine 123 staining,agarose gel electrophoresis,Hoechst 33258 staining,activity of caspase-3 and caspase-8 was detected,and B16 cell apoptosis induced by HB was observed.Results HB inhibited the B16 cell proliferation in vitro.The inhibitory effect was dose and time dependent,and the IC50 was 16.59,9.29 and 6.22 mg·L^-1 when B16 cell was exposed to HB for 24,48 and 72h,respectively.B16 cells treated with HB showed typical characteristics of apoptosis：decreased Rhodamine 123 fluorescence intensity,karyopyknosis and DNA Ladder.Caspase-3 and caspase-8 might be involved in this process.Conclusion HB could inhibit cell growth and induce apoptosis of B16 cells.