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 diseas...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.展开更多
基金Science and Technology Development Fund(FDCT)of Macao SAR(069/2015/A2134/2014/A3+4 种基金062-2017-AIR)Research Committee,University of Macao(MYRG2015-00182-ICMS-QRCMMYRG2015-00214-ICMSQRCMMYRG139(Y1-L4)-ICMS12-LMYand MYRG2016-00129-ICMS-QRCM)
文摘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.
