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 ofL.), 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 cytochromeactivation 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.

Absorption of euphobiasteroid across Caco-2 cell monolayer model / 中草药

Objective: To study the absorption and transportation of euphobiasteroid in Caco-2 cell monolayer model. Methods: Caco-2 cell monolayer model was used to study the process of bi-direction transport of euphobiasteroid, and the effects of time, drug concentration, and inhibitors on the process were investigated. The concentration of euphobiasteroid was detected by UPLC-MS/MS, and the apparent permeability coefficient (Papp) and apparent permeability (PDR) were calculated. Results: During the transport process of euphobiasteroid in Caco-2 cells, the Papp values of variety concentration were from 1 × 10-6 to 1 × 10-5. The cumulative transshipment volume was increased with time and concentration, which presented concentration-dependent manner. The PDR values of 10, 30, and 50 μmol/L euphobiasteroid were 1.35, 0.83, and 0.65, respectively. Verapamil hydrochloride could promote the transportation of euphobiasteroid from AP side to BL side. Conclusion: The absorption of euphobiasteroid in intestine is moderate and mainly through passive transport. There may be excretion mechanism of intestinal transport protein in the intestine absorption of euphobiasteroid.

Analysis and determination of diterpenoids in unprocessed and processed Euphorbia lathyris seeds by HPLC-ESI-MS / 药物分析学报

Euphorbia lathyris (Caper spurge) is a toxic and potent Chinese materia medica (T/PCMM).This study sought a method for identifying five diterpenoids (Euphorbia factors L1-L3,L7a and L8) with the spectra of UV and mass,quantifying three diterpenoids L1,L2,and L8 in crude extracts of unprocessed and processed E.lathyris seeds by liquid chromatography/electrospray ionization mass spectrometry (LC-ESI-MS).The analysis was achieved on an Agilent Eclipse XDB-C18 column (4.6 mm × 150 mm i.d.,5 μm) with an isocratic elution with a mobile phase consisting of water and acetonitrile at a flow rate of 0.25 mL/min at column temperature of 30 ℃ and UV detection was set at 272 nm.An ESI source was used with a positive ionization mode.The calibration curve was linear in the ranges of 9.9-79 μg/mL for Euphorbia factor L1,3.8-30.5 μg/mL for Euphorbia factor L2,and 1.0-20.6 μg/mL for Euphorbia factor L8.The average recoveries (n=6) of three diterpenoids were 98.39％,91.10％ and 96.94％,respectively,with RSD of 2.5％,2.4％ and 2.1％,respectively.The contents of the three diterpenoids in processed E.lathyris seeds were 3.435,1.367 and 0.286 mg/g,respectively,which decreased more sharply than those in unprocessed E.lathyris seeds which were 4.915,1.944 and 0.425 mg/g,respectively.The method is simple,accurate,reliable and reproducible,and it can be applied to control the quality of unprocessed and processed E.lathyris seeds.

Analysis and determination of diterpenoids in unprocessed and processed Euphorbia lathyris seeds by HPLC-ESI-MS / 药物分析学报

Euphorbia lathyris （Caper spurge） is a toxic and potent Chinese materia medica （T/PCMM）. This study sought a method for identifying five diterpenoids （Euphorbia factors LI-L3, L7a, and Ls） with the spectra of UV and mass, quantifying three diterpenoids L1, L2, and L8 in crude extracts of unprocessed and processed E. lathyris seeds by liquid chromatography/ electrospray ionization mass spectrometry （LC-ESI-MS）. The analysis was achieved on an Agilent Eclipse XDB-C18 column （4.6 mm× 150mm i.d., 5 μm） with an isocratic elution with a mobile phase consisting of water and acetonitrile at a flow rate of 0.25 mL/min at column temperature of 30 ℃ and UV detection was set at 272 nm. An ESI source was used with a positive ionization mode. The calibration curve was linear in the ranges of 9.9-79 μg/mL for Euphorbia factor Lb 3.8-30.5μg/mL for Euphorbia factor L2, and 1.0-20.6 μg/mL for Euphorbia factor LB. The average recoveries （n=6） of three diterpenoids were 98.39%, 91.10% and 96.94%, respectively, with RSD of 2.5%, 2.4% and 2.1%, respectively. The contents of the three diterpenoids in processed E. lathyris seeds were 3.435, 1.367 and 0.286 mg/g, respectively, which decreased more sharply than those in unprocessed E. lathyris seeds which were 4.915, 1.944 and 0.425 mg/g, respectively. The method is simple, accurate, reliable and reproducible, and it can be applied to control the quality of unprocessed and processed E. lathyris seeds.

The Inhibitory Effect of Phytoclear-EL1 on Melanogenesis

BACKGROUND: Phytoclear-EL1, an extract from Euphorbia lathyris seeds, has a whitening effect due to inhibition of tyrosinase activity. OBJECTIVE: The purpose of this study was to investigate the inhibitory effect of phytoclear-EL1 on melanogenesis. METHODS: Cultured B-16 melanoma cells and 30 human volunteers were used for in vitro and in vivo studies, respectively. Phytoclear-EL1 was added to the cultured B-16 melanoma cells, and applied to UVB-induced hyperpigmented lesions of human volunteers twice daily for 7 weeks. Changes in the number of B-16 melanoma cells, as well as changes in morphology, melanin content, and tyrosinase activity, were measured and then compared with the normal control and the 10(-3)M arbutin groups. Also, the effect of phytoclear-EL1 on UVB-induced hyperpigmented lesions was examined through subjective and objective measurements. RESULTS: In the in vitro study (p<0.05), the number, melanin content, and tyrosinase activity of cultured B-16 melanoma cells were decreased in the 5microgram/ml phytoclear-EL1 group compared to the control group. On objective assessment with a chromameter, the 0.2% phytoclear-EL1 group had a larger difference in the mean L values before and 7 weeks after applying phytoclear-EL1 as compared to the other groups. On subjective assessment by both the researchers and subjects 7 weeks after applying experimental materials, the 0.2% phytoclear-EL1 group and positive control (3% arbutin) had higher scores than the placebo groups. These results demonstrated that phytoclear-EL1 in vivo and in vitro had an inhibitory effect on melanogenesis. CONCLUSION: Phytoclear-EL1 may be a candidate extract in the control of hyperpigmentary disorders.

Studies on Chemical constituents in seeds of Euphorbia lathyris / 中草药

Objective To study the chemical constituents in the seeds of Euphorbia lathyris.MethodsCompounds were isolated by methods of column chromatography(silica gel,including reversed phase),Sephadex,and recrystallization.On the basis of spectroscopic methods including IR,MS,NMR,and X-ray,structures of compounds were confirmed.Results Twenty-two multi-type compounds were isolated from ethanol extract in the seeds of E.lathyris.Their structures were identified as 5,15-O-diacetyl-3-O-phenyl-6(17)-epoxylathyrol(1),5,15-O-diacetyl-3,7-O-dibenzoyl-7-hydroxylathyrol(2),5,15-O-diacetyl-3-O-benzoyl-lathyrol(3),20-O-hexadecanoyl-ingenol(4),3-O-hexadecanoyl-ingenol(5),15,17-O-diacetyl-3-O-cinnamoyl-17-hydroxyjolkinol(6),5,15,17-O-triacetyl-3-O-benzoyl-17-hydroxyisolathyrol(7),5,15-O-diacetyl-3-O-nicotinoyl-lathyrol(8),5,15-O-diacetyl-3-O-benzoyl-7-O-nicotinoyl-7-hydroxy-lathyrol(9),ingenol(10),lathyrol(11),esculetin(12),?-sitosterol(13),benzene-1,2,3-triol(14),palmiticacid(15),2,3-dihydroxypropyl icosanoate(16), 2,3-dihydroxypropyl oleate(17),2,3,4-trihydroxybutyl hexadec-3-enoate(18),aurantianide acetate(19),benzoic acid(20),p-hydroxybenzoic acid(21),oleic acid(22).Conclusion Among these,compounds 10,11,1419 are obtained from this plant for the first time and compounds 1-3 are the main diterpenes.