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SH-SY5Y 신경세포주에서 에탄올 처치 및 금단이 신호전달계 단백질 인산화에 미치는 영향

The Regulation of ERK, GSK3β and AKT after Acute Ethanol Exposure and Withdrawal in SH-SY5Y Human Neuroblastoma Cell-Line

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주제분류정신과학

초록

Objectives: This study aimed to demonstrate the activities and phosphorylation changes induced by acute ethanol treatment and withdrawal conditions in the intracellular signal transduction molecules [such as extracellular signal-regulated kinase (ERK), glycogen synthase kinase 3β(GSK3β), and Akt] of the SH-SY5Y neuroblastoma cell line. Methods: The acute treatment exposed SH-SY5Y cells to 100 mM ethanol, and we took samples 30 minutes, 60 minutes, and 24 hours after initiating this treatment. After 24 hours’ continuous ethanol treatment, we initiated ethanol withdrawal, taking samples at 30 minutes and 60 minutes. We assayed the kinase phosphorylations via an immunoblot analysis using phosphorspecific antibodies, quantified by optical densitometry. Results: Ethanol treatment induced a transient increase in phosphorylation of GSK3β and Akt at 30 minutes but failed to change the phosphorylation level of ERK. Ethanol withdrawal induced a transient ERK phosphorylation increase at 30 minutes, but it had no effect on the phosphorylation of GSK3β or Akt. Conclusion: The results indicate that the ethanol-induced cellular response includes the ERK,GSK3β, and Akt systems. In particular, the ERK pathway may play a role in the acute withdrawal response. This also suggests that a relatively short exposure to ethanol, such as the 24-hour exposure in this study, can induce functional adaptation within a cell.

Abstract

Objectives: This study aimed to demonstrate the activities and phosphorylation changes induced by acute ethanol treatment and withdrawal conditions in the intracellular signal transduction molecules [such as extracellular signal-regulated kinase (ERK), glycogen synthase kinase 3β(GSK3β), and Akt] of the SH-SY5Y neuroblastoma cell line. Methods: The acute treatment exposed SH-SY5Y cells to 100 mM ethanol, and we took samples 30 minutes, 60 minutes, and 24 hours after initiating this treatment. After 24 hours’ continuous ethanol treatment, we initiated ethanol withdrawal, taking samples at 30 minutes and 60 minutes. We assayed the kinase phosphorylations via an immunoblot analysis using phosphorspecific antibodies, quantified by optical densitometry. Results: Ethanol treatment induced a transient increase in phosphorylation of GSK3β and Akt at 30 minutes but failed to change the phosphorylation level of ERK. Ethanol withdrawal induced a transient ERK phosphorylation increase at 30 minutes, but it had no effect on the phosphorylation of GSK3β or Akt. Conclusion: The results indicate that the ethanol-induced cellular response includes the ERK,GSK3β, and Akt systems. In particular, the ERK pathway may play a role in the acute withdrawal response. This also suggests that a relatively short exposure to ethanol, such as the 24-hour exposure in this study, can induce functional adaptation within a cell.

참고문헌 (39)

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