苦参碱对Raji细胞凋亡及凋亡相关蛋白Fas/FasL表达的影响

目的：探讨中药苦参碱对人Burkitts淋巴瘤Raji细胞凋亡的作用及其可能机制.方法：应用MTT法测定苦参碱对Raji细胞生长的抑制作用;AnnexinⅤ-FITC/PI双染法检测细胞凋亡率;WesternBlot检测Fas,FasL,Caspase-3蛋白的表达量.结果：苦参碱0.2～1.6g/L作用Raji细胞24,48,72h后,对Raji细胞生长具有增殖抑制作用,且呈明显的量效和时效关系,48h半数抑制浓度（IC50）为1.34g/L.苦参碱0.4,0.8,1.6g/L组作用48h后,Raji细胞总凋亡率分别为15.10%,27.88%,48.08%,与对照组8.78%相比较差异具有统计学意义（P〈0.05,P〈0.01）.Western Blot检测结果显示Fas,FasL,Caspase-3蛋白表达水平随苦参碱浓度的提高而增加.结论：一定浓度苦参碱可诱导Raji细胞发生凋亡,其凋亡机制可能与上调Fas,FasL蛋白的表达以及激活Caspase-3有关.

Raji cell apoptosis-inducing effects of matrine and its effect on Fas/FasL protein expression

AIM：To invest the role of matrine-induced apoptosis of Human Burkitt＇s lymphoma Raji cells and its mechanism.METHODS：The proliferation inhibition rate of Raji cells was performed by MTT analysis.Cell apoptosis rate was detected by AnnexinⅤ-FITC/PI double staining method.The Fas,FasL and Caspase-3 protein expresssion of Raji cells treated with matrine were evaluated by Western blot.RESULTS：After Raji cells were cocultured with matrine 0.2-1.6 g/L for 24,48,72 h in vitro,the cell proliferation was inhibited in time-and dose-dependent manner,its half maximal inhibitory concentration（IC50）of 48 h was 1.34 g/L.When the Raji cells were cultured for 48 h by matrine 0.4,0.8,1.6 g/L,the total apoptosis rate 15.10%,27.88%,48.08% detected by AnnexinⅤ-FITC/PI double staining,had statistical significance compared with control group 8.78%,（P〈0.05 or P〈0.01）.The protein expresssion levels of Fas,Fasl and Caspase3 increased after Raji cells were treated by matrine0.4,0.8,1.6 g/L with a concentration-effect relationship.CONCLUSION：Matrine can induce the apoptosis of Raji cells in vitro,one of its possible mechanisms is that matrine activates caspase-3 through upregulation of Fas and FasL.