单用雷公藤内酯醇及联合硼替佐米对霍奇金淋巴瘤细胞株作用的对照研究

The effect of triptolide （TPL） and triptolide combined with PS--341 on Hodgkin＇s disease cells

目的 观察单用雷公藤内酯醇（TPL）及联合硼替佐米（PS-341）对霍奇金淋巴瘤细胞株L428的作用及机制探讨.方法 采用MTT比色法检测TPL、PS-341对L428细胞株体外生长的抑制作用;并予Annexin V和PI双染色,周期分析检测细胞凋亡;同时采用Western-blot法检测PS-341、TPL对L428细胞Bcl-2、Bax和Caspase-3、9及PARP蛋白表达水平的影响.结果 TPL可明显抑制L428细胞增殖,且生长抑制率与药物作用浓度呈正相关,以PS-341（80 ng/m1）联合TPL5～500 ng/ml处理L428细胞24h时的抑制作用优于单用TPL的作用;PI的DNA染色及AnnexinV-PI双标记法结果显示：细胞凋亡率与TPL作用浓度呈正相关;TPL作用L428细胞后,随浓度的增加,Bcl-2蛋白表达下调,而Bax蛋白表达上调;Caspase-3、Caspase-9和PARP蛋白均可出现明显的剪切带.结论 （1）TPL可能通过线粒体途径和改变Bcl-2家族抗凋亡因子和促凋亡因子的蛋白表达比例,诱导L428细胞凋亡;（2）PS-341能增强TPL诱导L428细胞凋亡的作用.

Objective To observe the effect of Triptolide（TPL） and Triptolide combined with PS-341 on hodgkin＇s disease CellsMethods This study was performed on cultured L428 cells by using MTT studies,the immersion objective,flow cytometry with Annexin-V/PI.Results The result of MTT colourimetry showed ： the proliferation of L428 dealed with TPL for 24h,48h.72h could markly be inhibited ,and the growing inhibition ratio was positive correlation with the concentration of the drug,showing concentration dependence,and the inhibition effect of TPL combined with PS-341 was much more than the effect of TPL on L428 cells ,after L428 was dealed with TPL,the level of Bcl-2 protein was down regulation with the increasing of the concentration of drug,but the level of bax was up regulation;the shearing band were showed up in caspase-3,caspase-8 and PARP protein. Conclusion （1）TPL can induce the apoptosis of L428,and the study shows the apoptosis of L428 is induced by the chondriosome pathway; （2）The study shows a synergistic depressant effect between TPL and PS-341,for the cytotoxic effects of TPL on L428 can be enhanced by combining PS-341.