RBL-2H3 세포에서 Fluoxetine에 의한 Tryptophan Hydroxylase 활성도 변화 = Fluoxetine-induced Changes on Activity of Tryptophan Hydroxylase at RBL-2H3 Cells

다국어 초록 (Multilingual Abstract)

Objectives : The aim of the present study is to explore the effect of fluoxetine on transcription, translation and activity of tryptophan hydroxylase (TPH), and intracellular level of serotonin.
Methods : The expression level of the TPH mRNA and the protein, the TPH enzyme activity, and the intracellular level of serotonin were explored at the fluoxetine-treated RBL-2H3 cells. Real-time RT-PCR and immunoblotting analysis confirmed changes in the expression of TPH mRNA and protein. The activity of TPH was measured using [3H]tryptophan. The intracellular level of serotonin was measured by HPLC.
Results : The TPH activity was gradually increased on time from 24hr to 72hr. The real-time RT-PCR also revealed that the TPH mRNA was increased at 12, 24 and 72hr in the fluoxetine-treated RBL-2H3 cells. The immunoblotting analysis also revealed that the TPH protein was decreased at 72hr in the fluoxetine-treated RBL-2H3 cells. The intracellular level of serotonin was increased at 48hr after treatment of fluoxetine.
Conclusions : Fluoxetine induced the increases of the TPH mRNA, the TPH enzyme activity and intracellular level of serotonin, and the decrease of the TPH protein expression at the RBL-2H3 cells.

국문 초록 (Abstract)

목 적：본 연구는 fluoxetine이 TPH 전사, 효소 활성, 및 단백질 생성, 그리고 세포 내 세로토닌 농도 변화에 어떠한 영향을 미치는지 알아보고자 실시하였다. 방 법：Fluoxetine이 처리 된 RBL-2H3 세포에서 Fluoxetine에 의한 Tryptophan Hydroxylase 활성도 변화 Korean J Psychopharmacol 2004;15(4):449-456 456 TPH 효소 활성, mRNA 및 단백질 수준, 그리고 세포 내 세로토닌 농도 수준에 대해 알아보았다. TPH 효소 활성은 [3H]tryptophan을 이용하여 효소 반응에 의해 생산되어지는 [3H]H2O로 측정되었다. TPH mRNA 및 단백질 변화는 real-time RT-PCR 및 면역 탁본법(imu -nobloting)에 의해 분석되었고, 세포 내 세로토닌 농도는 HPLC를 이용하여 UV 280 ㎚에서 측정되었다. 결 과：TPH 효소 활성은 fluoxetine 처리 24시간부터 72시간까지 시간에 따라 점차 증가하여 72시간에 최대가 되었다. Real-time RT-PCR에 의해 확인 된 TPH mRNA는 12, 24 및 72시간에서 증가되었고 12시간에 최대가 되었다. 48시간에는 변화가 없었다. 면역 탁본법에 의해 72시간에 TPH 단백질의 감소가 확인되었고, 세포 내 세로토닌 농도는 fluoxetine 처리 48시간에 증가되었다.
결 론：Fluoxetine은 RBL-2H3 세포에서 TPH 효소 활성, mRNA 발현 및 세로토닌 농도 증가를 유도하였고, TPH 단백질 발현은 감소시켰다.

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