Inhibition of miR-873 provides therapeutic benefit in lipopolysaccharide-induced Parkinson disease animal model

OBJECTIVE Neuroinflammation plays a critical role in neurodegenerative disorders,although the inflammation may not the initiating factor.Parkinson disease(PD)is characterized pathologically by the accumulation of alpha synuclein(α-syn)and the loss of the dopamine(DA)neurons in the substantia nigra(SN),which has been reported to be induced by the stereotaxic injection of lipopolysaccharide(LPS)to the SN region in rodents.This study is to investigate the therapeutic benefit of the inhibition of miR-873 in PD.METHODS Rats received the right-unilaterally injection with concentrated LV-sponge or LV-EGFP 3 d before LPS treatment,7 or 14 d after LPS treatment.The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 21 d after LPS injection.The regulation of miR-873 on the genes related with cholesterol transport and inflammation was assayed in SH-SY5Y cells and U251 cells.RESULTS TLR4-My D88 signaling pathway was involved the regulation of miR-873 by LPS.The luciferase assay showed that HMGCR,ABCA1 and A20 were down-stream genes of miR-873.The transfection of miR-873 decreased the cholesterol levels in cell membrane,but increased in lysosome in SH-SY5Y cells.Compared with the control SH-SY5Y cells,cholesterol levels were higher in lysosome withα-synuclein overexpression or LPS treatment.The transfection of miR-873 increased theα-syn levels in lysosome in cel s withα-synuclein overexpression.The loss of dopaminergic neuorns induced by LPS was significantly respectively decreased by 22.8%,35.6%and 57% after the inhibition of miR-873 at 3 d before LPS treatment,7 or14 d after LPS treatment.Compared with LPS-treated group,the number of the rotation of rats was decreased by 60.4%,33.5%and 13.2%after the inhibition of miR-873 at 3 d before LPS treatment,7or 14 d after LPS treatment.The inhibition of miR-873 significantly decreased accumulation ofα-syn.The m RNA levels of HMGCR,ABCA1 and A20 in SN were decreased by LPS treatment,which was attenuated by the injection of LV-sponge.CONCLUSION The selective regulation of miR-873 can protect the dopaminergic neurons from the LPS-induced damage.The inhibition of miR-873 can attenuate the relocation of cholesterol in lysosome and the accumulation ofα-syn in neurons induced by LPS via the regulation of HMGCR,ABCA1 and A20.

Protective role of brain CYP2J in diverse Parkinson disease animal models

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.