title: P2X1 Receptor-Mediated Ca2+Influx Triggered by DA-9801 Potentiates Nerve Growth Factor-Induced Neurite Outgrowth.
abstract: Nerve growth factor (NGF)-induced neuronal regeneration has emerged as a strategy to treat neuronal degeneration-associated disorders. However, direct NGF administration is limited by the occurrence of adverse effects at high doses of NGF. Therefore, development of a therapeutic strategy to promote the NGF trophic effect is required. In view of the lack of understanding of the mechanism for potentiating the NGF effect, this study investigated molecular targets of DA-9801, a well-standardized Dioscorea rhizome extract, which has a promoting effect on NGF. An increase in intracellular calcium ion level was induced by DA-9801, and chelation of extracellular calcium ions with ethylene-bis(oxyethylenenitrilo)tetraacetic acid (EGTA) suppressed the potentiating effect of DA-9801 on NGF-induced neurite outgrowth. In addition, EGTA treatment reduced the DA-9801-induced phosphorylation of extracellular signal-regulated kinase1/2 (ERK1/2), the major mediators of neurite outgrowth. To find which calcium ion-permeable channel contributes to the calcium ion influx induced by DA-9801, we treated PC12 cells with various inhibitors of calcium ion-permeable channels. NF449, a P2X1 receptor selective antagonist, significantly abolished the potentiating effect of DA-9801 on NGF-induced neurite outgrowth and abrogated the DA-9801-induced ERK1/2 phosphorylation. In addition, transfection with siRNA of P2X1 receptor significantly reduced the DA-9801-enhanced neurite outgrowth. In conclusion, calcium ion influx through P2X1 receptor mediated the promoting effect of DA-9801 on NGF-induced neurite outgrowth via ERK1/2 phosphorylation.
authors.affiliation: Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University ; 84 Heukseok-ro, Dongjak-Ku, Seoul 06974, Republic of Korea.
authors.forename: Moon Jung
authors.initials: MJ
authors.lastname: Back
authors.affiliation: Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University ; 84 Heukseok-ro, Dongjak-Ku, Seoul 06974, Republic of Korea.
authors.forename: Hae Kyung
authors.initials: HK
authors.lastname: Lee
authors.affiliation: Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University ; 84 Heukseok-ro, Dongjak-Ku, Seoul 06974, Republic of Korea.
authors.forename: Joo Hyun
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University ; 84 Heukseok-ro, Dongjak-Ku, Seoul 06974, Republic of Korea.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Fu
authors.affiliation: Department of Research Planning & Management, Research Center of Dong-A ST Co., Ltd. ; 21 Geumhwa-ro, 105 beon-gil, Giheung-gu, Yongin-si, Gyeonggi-do 446-905, Republic of Korea.
authors.forename: Mi Won
authors.initials: MW
authors.lastname: Son
authors.affiliation: Department of Research Planning & Management, Research Center of Dong-A ST Co., Ltd. ; 21 Geumhwa-ro, 105 beon-gil, Giheung-gu, Yongin-si, Gyeonggi-do 446-905, Republic of Korea.
authors.forename: Sang Zin
authors.initials: SZ
authors.lastname: Choi
authors.affiliation: Department of Research Planning & Management, Research Center of Dong-A ST Co., Ltd. ; 21 Geumhwa-ro, 105 beon-gil, Giheung-gu, Yongin-si, Gyeonggi-do 446-905, Republic of Korea.
authors.forename: Hyo Sang
authors.initials: HS
authors.lastname: Go
authors.affiliation: Department of Biomedical Sciences, Korea University College of Medicine , Seoul 136-705, Republic of Korea.
authors.forename: Sungjae
authors.initials: S
authors.lastname: Yoo
authors.affiliation: Department of Biomedical Sciences, Korea University College of Medicine , Seoul 136-705, Republic of Korea.
authors.forename: Sun Wook
authors.initials: SW
authors.lastname: Hwang
authors.affiliation: Department of Environmental and Health Chemistry, College of Pharmacy, Chung-Ang University ; 84 Heukseok-ro, Dongjak-Ku, Seoul 06974, Republic of Korea.
authors.forename: Dae Kyong
authors.initials: DK
authors.lastname: Kim
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D065096:Calcium Chelating Agents / Q000494:pharmacology; D002413:Cations, Divalent / Q000378:metabolism; D004533:Egtazic Acid / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D020932:Nerve Growth Factor / Q000378:metabolism*; D016501:Neurites / Q000187:drug effects* / Q000502:physiology; D000071446:Neuronal Outgrowth / Q000187:drug effects* / Q000502:physiology; D018696:Neuroprotective Agents / Q000494:pharmacology; D016716:PC12 Cells; D010766:Phosphorylation / Q000187:drug effects; D028321:Plant Preparations / Q000494:pharmacology*; D058905:Purinergic Agents / Q000494:pharmacology*; D034741:RNA, Small Interfering; D051381:Rats; D058474:Receptors, Purinergic P2X1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D065096:Calcium Chelating Agents; D002413:Cations, Divalent; C562027:DA-9801; D018696:Neuroprotective Agents; D028321:Plant Preparations; D058905:Purinergic Agents; D034741:RNA, Small Interfering; D058474:Receptors, Purinergic P2X1; D004533:Egtazic Acid; D020932:Nerve Growth Factor; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002118:Calcium
keywords: DA-9801; P2X1; calcium ion influx; neurite outgrowth