摘要
OBJECTIVE To investigate the effects of T-006(tetramethylpyrazine derivative)in promotingα-Synuclein(α-Syn)degradation and evaluated the neuroprotective effects in cellular and animalα-Syn model of Parkinson disease(PD).METHODS The inducible PC12 cells overexpressingα-syn and the homozygous transgenic(Tg)mice expressing A53T humanα-syn were used to evaluate the neuroprotective effects of T-006.For cellular study,MTT,Western blotting,proteasomal activity assay and qRT-PCR were applied to analyze the pharmacological effects and underlying mecha⁃nisms.The gene knock-down and overexpression approaches were used to dissect the molecular signaling pathways.For animal study,ten-month-old homozygousα-Syn Tg mice were treated with T-006(3 mg·kg-1)daily by gavage for four weeks.The Western blotting,immunohistochemistry and behavioral tests were applied to determine the neuropatho⁃logical changes.RESULTS T-006 promoted the degradation of WT and mutantα-Syn in PC12α-Syn inducible cells via an ubiquitin-proteasome system(UPS)dependent and autophagy-lysosome pathway independent manner.The mecha⁃nism of action involved the upregulation of 20S proteasome subunit LMP7 expression,which leads to activation of the chymotrypsin-like proteasomal activity for protein degradation.Mechanistically,we demonstrated that T-006 activated PKA/Akt/mTOR pathway upstream for LMP 7 up-regulation and UPS activation.Finally,we illustrated that T-006 promoted both Triton-soluble and-insoluble forms ofα-syn and protected againstα-Syn-induced neurotoxicity in A53Tα-Syn Tg mice.CONCLUSION T-006 is a potent UPS activator which promotes the degradation of pathogenic proteinα-Syn in cellular and animal PD models.Our study thus high-lights the therapeutic potential of small molecular UPS activator like T-006 in the treatment of PD and related conditions.
OBJECTIVE To investigate the effects of T-006（tetramethylpyrazine derivative） in promoting α-Synuclein（α-Syn） degradation and evaluated the neuroprotective effects in cellular and animal α-Syn model of Parkinson disease（PD）. METHODS The inducible PC12 cells overexpressing α-syn and the homozygous transgenic（Tg） mice expressing A53 T human α-syn were used to evaluate the neuroprotective effects of T-006. For cellular study, MTT, Western blotting,proteasomal activity assay and q RT-PCR were applied to analyze the pharmacological effects and underlying mechanisms. The gene knock-down and overexpression approaches were used to dissect the molecular signaling pathways.For animal study, ten-month-old homozygous α-Syn Tg mice were treated with T-006(3 mg·kg-1) daily by gavage for four weeks. The Western blotting, immunohistochemistry and behavioral tests were applied to determine the neuropathological changes. RESULTS T-006 promoted the degradation of WT and mutant α-Syn in PC12 α-Syn inducible cells via an ubiquitin-proteasome system（UPS） dependent and autophagy-lysosome pathway independent manner. The mechanism of action involved the upregulation of 20 S proteasome subunit LMP7 expression, which leads to activation of the chymotrypsin-like proteasomal activity for protein degradation. Mechanistically, we demonstrated that T-006 activated PKA/Akt/mT OR pathway upstream for LMP 7 up-regulation and UPS activation. Finally, we illustrated that T-006 promoted both Triton-soluble and-insoluble forms of α-syn and protected against α-Syn-induced neurotoxicity in A53 T α-Syn Tg mice. CONCLUSION T-006 is a potent UPS activator which promotes the degradation of pathogenic protein α-Syn in cellular and animal PD models. Our study thus high-lights the therapeutic potential of small molecular UPS activator like T-006 in the treatment of PD and related conditions.
出处
《中国药理学与毒理学杂志》
CAS
北大核心
2019年第9期658-659,共2页
Chinese Journal of Pharmacology and Toxicology
基金
Science and Technology Development Fund(FDCT)of Macao SAR(069/2015/A2
134/2014/A3
062-2017-AIR)
Research Committee,University of Macao(MYRG2015-00182-ICMS-QRCM
MYRG2015-00214-ICMSQRCM
MYRG139(Y1-L4)-ICMS12-LMY
and MYRG2016-00129-ICMS-QRCM)
