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Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.
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AIM:To determine the contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae and Semen Euphorbiae Pulveratun from different habitats.METHODS:A kromasil C_(18) column(150 mm×4.6 mm,5 μm)was used with methanol-water(68:32)as the mobile phase and UV detection was at 280 nm.RESULTS:The linear ranges of euphorbiasteroid and lathyrol diadetate benzozte were from 0.200 μg to 1.200 μg and 0.206 μg to 1.651 μg(r = 0.999 7,r = 0.999 9).The contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae from different habitats ranged from 0.450 8% to 0.625 1 % and 0.692 3% to 0.752 9%.The contents of two kinds of compositions in Semen Euphorbiae Pulveratun ranged from 0.156 8% to 0.221 1% and 0.232 2% to 0.283 0%.CONCLUSION:The method for quantitating euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae is accurate and reliable,and can be used to control the quality of toxicity Semen Euphorbiae and Semen Euphorbiae Pulveratun.
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AIM:To determine the contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae and Semen Euphorbiae Pulveratun from different habitats.METHODS:A kromasil C18 column(150 mm?4.6 mm,5 ?m) was used with methanol-water(68 :32) as the mobile phase and UV detection was at 280 nm.RESULTS:The linear ranges of euphorbiasteroid and lathyrol diadetate benzozte were from 0.200 ?g to 1.200 ?g and 0.206 ?g to 1.651 ?g(r=0.999 7,r=0.9999).The contents of euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae from different habitats ranged from 0.450 8% to 0.625 1% and 0.692 3% to 0.752 9%.The contents of two kinds of compositions in Semen Euphorbiae Pulveratun ranged from 0.156 8% to 0.221 1% and 0.232 2% to 0.283 0%.CONCLUSION:The method for quantitating euphorbiasteroid and lathyrol diadetate benzozte in Semen Euphorbiae is accurate and reliable,and can be used to control the quality of toxicity Semen Euphorbiae and Semen Euphorbiae Pulveratun.