OBJECTIVE Cytochrome P450(CYP)2J2 is highly expressed in many kinds of human tumors and promotes tumor cell growth via regulating the metabolism of arachidonic acids.The purposes of this study were toidentify the new ...OBJECTIVE Cytochrome P450(CYP)2J2 is highly expressed in many kinds of human tumors and promotes tumor cell growth via regulating the metabolism of arachidonic acids.The purposes of this study were toidentify the new inhibitor of CYP2J2 from natural compounds and evaluate its potential to inhibit hepatoma carcinoma cells.METHODS Total fifty natural products were screened for the inhibitory potency against the activity of CYP2J2-mediated astemizole O-demethylation via LCMS/MS analysis.Enzyme kinetic and molecular docking studies were also carried out.RESULTS Our data found that plumbagin potently inhibited CYP2J2 with IC50value at 3.42,3.37 and 1.17μmol·L-1in rat liver microsomes,human liver microsomes(HLMs)and recombinant CYP2J2(rC YP2J2),respectively.Further enzyme kinetic studies showed that plumbagin was a mixed-type inhibitor of CYP2J2 in HLMs and r CYP2J2 with Kivalues of 1.88and 0.92μmol·L-1,respectively.Docking data presented that plumbagin interacted with CYP2J2 mainly through GLU222 and ALA223,which were crucial residues for substrates binding.At the same time,plumbagin showed cytotoxicity effects on hepatic carcinoma cell lines,such as HepG 2 and SMMC-7721,with IC50values at 11.55±1.06and(13.15±1.11)μmol·L-1,respectively.CONCLUSION These results indicated that plumbagin was a potent CYP2J2 inhibitor and potential anticancer agent.Further studies are needed to cover the mechanism of its antitumor activity.展开更多
OBJECTIVE CYP2 family including CYP2C and CYP2J is the predominant arachidonic acid(AA)epoxygenase,and the epoxidation of AA produces four regioisomeric cis-epoxyeicosatrienoic acids(5,6-,8,9-,11,12-,and 14,15-EET).Hu...OBJECTIVE CYP2 family including CYP2C and CYP2J is the predominant arachidonic acid(AA)epoxygenase,and the epoxidation of AA produces four regioisomeric cis-epoxyeicosatrienoic acids(5,6-,8,9-,11,12-,and 14,15-EET).Human CYP2J2 is one of the main CYP isoforms expressed in brain,but CYP2C8 was present at a low level.The aim of this study is to investigate the roles of brain CYP2J in Parkinson disease.METHODS Rats received the right-unilateral y injection with concentrated LV-CYP2J3 or LV-EGFP in the substantia nigra(SN)at 3 d before LPS or 6-OHDA treatment.The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 14 and 21 d after LPS injection.The influence of CYP2J-dependent derivative,14,15-EET,on the genes related with oxidative stress was assayed in SH-SY5Y cells.RESULTS CYP2J overexpression or 14,15-EET treatment significantly increased the levels of SOD1,CAT,GPX1,NRF2 and KEAP1 in neurons.TLR4-My D88 signaling pathway was involved the down-regulation of CYP2J by LPS.The binding of p-CREB with the promoter of CYP2J was inhibited by the LPS treatment.The loss of dopaminergic neurons in the right SN induced by LPS or 6-OHDA was significantly decreased by CYP2J3 transfection at 21 d after LPS injection.Compared with LPS or 6-OHDA group,the number of the rotation of rats was decreased by 42.6% and 60.7%by CYP2J3 transfection at 14 d after LPS or 6-OHDA injection;meanwhile,the rotation number was decreased by 12.7%and 21.3%at 21 d.The accumulation of alpha synuclein induced by LPS was significantly decreased by CYP2J3 transfection.The mR NA levels of SOD1,CAT,GPX1,NRF2 and KEAP1 in SN were decreased by LPS,which was attenuated by the injection of LV-CYP2J3.CONCLUSION Brain CYP2J can play a protective role in the damage of the inflammation and oxidative stress to the dopaminergic neurons.Brain CYP2J-dependent derivatives from AA may have therapeutic effects in Parkinson disease via the up-regulation of the antioxidant system in neurons.展开更多
基金The project supported by National Natural Science Foundation of China(81301908)the Science and Technology Commission of Shanghai Municipality(15140904700,13ZR1412600 and 14DZ2270100)
文摘OBJECTIVE Cytochrome P450(CYP)2J2 is highly expressed in many kinds of human tumors and promotes tumor cell growth via regulating the metabolism of arachidonic acids.The purposes of this study were toidentify the new inhibitor of CYP2J2 from natural compounds and evaluate its potential to inhibit hepatoma carcinoma cells.METHODS Total fifty natural products were screened for the inhibitory potency against the activity of CYP2J2-mediated astemizole O-demethylation via LCMS/MS analysis.Enzyme kinetic and molecular docking studies were also carried out.RESULTS Our data found that plumbagin potently inhibited CYP2J2 with IC50value at 3.42,3.37 and 1.17μmol·L-1in rat liver microsomes,human liver microsomes(HLMs)and recombinant CYP2J2(rC YP2J2),respectively.Further enzyme kinetic studies showed that plumbagin was a mixed-type inhibitor of CYP2J2 in HLMs and r CYP2J2 with Kivalues of 1.88and 0.92μmol·L-1,respectively.Docking data presented that plumbagin interacted with CYP2J2 mainly through GLU222 and ALA223,which were crucial residues for substrates binding.At the same time,plumbagin showed cytotoxicity effects on hepatic carcinoma cell lines,such as HepG 2 and SMMC-7721,with IC50values at 11.55±1.06and(13.15±1.11)μmol·L-1,respectively.CONCLUSION These results indicated that plumbagin was a potent CYP2J2 inhibitor and potential anticancer agent.Further studies are needed to cover the mechanism of its antitumor activity.
基金supported by New Century Excellent Talents in University(NCET-11-0399)National Natural Science Foundation of China(81673503)
文摘OBJECTIVE CYP2 family including CYP2C and CYP2J is the predominant arachidonic acid(AA)epoxygenase,and the epoxidation of AA produces four regioisomeric cis-epoxyeicosatrienoic acids(5,6-,8,9-,11,12-,and 14,15-EET).Human CYP2J2 is one of the main CYP isoforms expressed in brain,but CYP2C8 was present at a low level.The aim of this study is to investigate the roles of brain CYP2J in Parkinson disease.METHODS Rats received the right-unilateral y injection with concentrated LV-CYP2J3 or LV-EGFP in the substantia nigra(SN)at 3 d before LPS or 6-OHDA treatment.The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 14 and 21 d after LPS injection.The influence of CYP2J-dependent derivative,14,15-EET,on the genes related with oxidative stress was assayed in SH-SY5Y cells.RESULTS CYP2J overexpression or 14,15-EET treatment significantly increased the levels of SOD1,CAT,GPX1,NRF2 and KEAP1 in neurons.TLR4-My D88 signaling pathway was involved the down-regulation of CYP2J by LPS.The binding of p-CREB with the promoter of CYP2J was inhibited by the LPS treatment.The loss of dopaminergic neurons in the right SN induced by LPS or 6-OHDA was significantly decreased by CYP2J3 transfection at 21 d after LPS injection.Compared with LPS or 6-OHDA group,the number of the rotation of rats was decreased by 42.6% and 60.7%by CYP2J3 transfection at 14 d after LPS or 6-OHDA injection;meanwhile,the rotation number was decreased by 12.7%and 21.3%at 21 d.The accumulation of alpha synuclein induced by LPS was significantly decreased by CYP2J3 transfection.The mR NA levels of SOD1,CAT,GPX1,NRF2 and KEAP1 in SN were decreased by LPS,which was attenuated by the injection of LV-CYP2J3.CONCLUSION Brain CYP2J can play a protective role in the damage of the inflammation and oxidative stress to the dopaminergic neurons.Brain CYP2J-dependent derivatives from AA may have therapeutic effects in Parkinson disease via the up-regulation of the antioxidant system in neurons.
