Euphorbia factor L2 induces apoptosis in A549 cells through the mitochondrial pathway

Euphorbia factor L2, a lathyrane diterpenoid isolated from caper euphorbia seed(the seeds of Euphorbia lathyris L.), has been traditionally applied to treat cancer. This article focuses on the cytotoxic activity of Euphorbia factor L2 against lung carcinoma A549 cells and the mechanism by which apoptosis is induced. We analyzed the cytotoxicity and related mechanism of Euphorbia factor L2 with an MTT assay, an annexin V-FITC/PI test, a colorimetric assay, and immunoblotting. Euphorbia factor L2 showed potent cytotoxicity to A549 cells. Euphorbia factor L2 led to an increase in reactive oxygen species(ROS) generation,a loss of mitochondrial electrochemical potential, release of cytochrome c, activation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase, suggesting that Euphorbia factor L2 induced apoptosis through a mitochondrial pathway. The cytotoxic activity of Euphorbia factor L2 in A549 cells and the related mechanisms of apoptotic induction provide support for the further investigation of caper euphorbia seeds.

LC-MS/MS同时检测大鼠血浆中千金子抗肿瘤活性成分及其药代动力学研究

目的建立灵敏快速的高效液相色谱-串联质谱法(LC-MS/MS),考察千金子中抗肿瘤活性成分大戟因子L2、L3口服(ig)、腹腔注射(ip)和尾静脉注射(iv)给药后在大鼠体内的药代动力学特征。方法采用LC-MS/MS内标法测定大戟因子L2、L3血药浓度,流动相为甲醇-水(8020),色谱柱为BSD Hypersil C_(18)柱(100 mm×2.1 mm,3μm),柱温为30℃,体积流量为0.2 mL/min,进样体积为5μL。质谱条件:电喷雾离子化(ESI)源,正离子模式扫描,多反应监测模式(MRM)检测各有效成分。采用WinNonlin 4.1药动学软件计算药动学参数。结果大戟因子L2和L3在5~1000 ng/mL范围内呈现出良好的线性关系(r^(2)>0.999),提取回收率为90.5%~107.2%,基质效应分别为89.1%~94.2%和88.1%~93.9%,日内、日间精密度RSD均<12.8%。大鼠尾静脉注射大戟因子L2、L3后,C_(max)、AUC_(0-24 h)分别为(20.76±7.55)、(13.48±4.59)μg/mL和(8.43±2.34)、(5.13±1.38)h·μg^(-1)·mL^(-1);大鼠腹腔注射注射大戟因子L2、L3后,C_(max)、AUC_(0-24 h)分别为(0.95±0.26)、(0.27±0.06)μg/mL和(3.52±0.90)、(0.98±0.29)h·μg^(-1)·mL^(-1);口服给药大戟因子L2、L3后,多个样品浓度因低于检测限而未检测到。结论建立的相关检测方法灵敏、快速、选择性好,可用于大鼠血浆中大戟因子L2、L3的同时测定及其药代动力学研究;药代动力学参数表明,大戟因子L2、L3口服吸收利用度较差,腹腔注射给药相同剂量下大戟因子L2在大鼠体内的暴露量较L3高。