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GH_3 세포(rat somatomammotropic tumor cell)에서 TRH가 Phospholipase D 활성에 미치는 효과
김동선,김태화,이창범,안유헌,윤미섭,한중수 대한내분비학회 2002 Endocrinology and metabolism Vol.17 No.4
연구배경: GH_3 세포는 TRH에 반응하여 세포막의 수용체와 G 단백, PI-PLC, PKC를 활성시켜 성장호르몬 및 프로락틴을 분비한다고 알려져 있다. PLD는 phosphatidylcholine을 phosphatidic acid(PA)와 choline으로 가수분해하는 효소로서 세포의 증식과 호르몬 분비에 관여한다. 본 연구는 GH_3 세포에서 아직 알려지지 않은 TRH의 PLD 활성에 대한 영향을 규명하고자 하였다. 방법: GH_3 세포를 1.5×10^6씩 분주하고 [^3H] myristate로 표지한 다음에 0.3% 알코올로 전 처치하였다. TRH 등의 시험제를 처치한 후에 메탄올로 반응을 종결하고 세포에서 총 지질을 추출하였다. PLD 활성은 박층크로마토그래피를 이용하여 총 [^3H] phospholipid에서 [^3H] phosphatidylethanol의 비율로 구하였다. 결과: TRH (1μM)의 처치 시에 PLC 활성은 44배 증가하였다. PLD 활성은 TRH (1μM), mastoparan (5μM), PMA(500nM)를 30분간 처치 시에 각각 1.9, 1.5, 2.2배 증가하였다. TRH(1μM)의 시간에 따른 PLD 활성 변동은 15, 30, 60, 120, 240분에 각각 142%, 170%, 172%, 160%, 115%의 증가를 보였다. 결론: GH_3 세포에서 TRH의 호르몬 분비와 세포증식의 신호전달 기전으로서는 PLC 활성뿐 아니라 PLD의 활성도 관여함을 시사한다. Backgroud: GH_3 cells are a well characterized and widely used model used for the in vitro study of growth hormone (GH) secretion. Thyrotropin releasing hormone (TRH) binds to receptors belonging to the family of G protein-coupled receptors, and secrets both GH & prolactin. Phospholipase D (PLD) is and enzyme that hydrolyses phosphatidylcholine to yield phosphatidic acid and choline, and plays important roles in cellular proliferation and hormonal secretion. To elucidate the pathway of the action of TRH in GH_3 cells, we investigated the activities of PLC and PLD in GH_3 cells treated with TRH or phorbor 12-myristate 13-acetate (PMA). Methos : GH_3 cells were labeled with [^3H] myristate, followed by incubation of with 0.3% ethanol, prior to before the addition of the agonists. The total lipids were extracted from the harvested cells following treatment with the agonists. The PLD activity was assessed by measuring [^3H] phosphatidylethanol from the [^3H] phospholipid using thin layer chromatography. Results : TRH (1μM) stimulated the PLD activity by 44-fold over that of the control values. TRH (1μM), mastoparan (5μM), and PMA (500μM) for 30 minutes increased PLD activity by 1.9, 1.5 and 2.2 fold, respectively, in comparison to the controls, The PLD activities after 15, 30, 60, 120 and 240 min treatments of TRH (1μM) were 142%, 170%, 172%, 160% and 115%, respectively. Conclusion : These results suggest that TRH stimulates not only PLC activity, but also the PLD activity in GH_3 cells (J Kor Soc Endocrinol 17:465∼472, 2002).
Propylthiouracil 투여에 의한 갑상선 기능저하 흰쥐 조직에서 알코올 탈수소효소 활성의 변화
백상현,김동선,이창범,박용수,안유헌,김태화,기춘석,강주섭 한양대학교 의과대학 2001 한양의대 학술지 Vol.21 No.2
Background: Thyroid hormone has been known to affect hepatic alcohol dehydrogenase (ADH) activity. Although the liver is the principal site of ethanol metabolism, stomach is also responsible in part for ethanol oxidation. The effects of thyroid hormone on ADH activity in gastric mucosa and other tissues of rats had not been previously examined. Method: The effects of thyroid hormone on liver, stomach, lung, and kidney ADH activities (nM of NADH/min/mg of cytosolic protein) have been investigated in male Sprague Dawley rats treated with propylthiouracil (50 mg/kg) for 14 days. Results: Whereas hepatic ADH activities were not changed by treatment with PTU(42.9(8.6 vs 45.2 (10.1), gastric ADH activities in PTU-treated rats increased by 258.8% of control rat (6.3 ( 0.6 vs 2.2 ( 1.2, p〈0.001). In the activities of other tissues, PTU treatment decreased lung ADH activity by 59.7% of control, and increased kidney ADH activities by 247.1% of control rats. Conclusion: These data suggest that hypothyroidism causes an increase of gastric alcohol metabolism, and that the increase of gastric ADH activity can partly restore the first pass metabolism of ethanol in hypothyroid rats.
지능박약 환자에서 Carbamazepine에 의해 발생한 골연화증 1예
김성훈,김동선,안유헌,최낙원,이창범,박용수,최웅환,김태화,김희수,백상현 대한내분비학회 2000 Endocrinology and metabolism Vol.15 No.2
Anticonvusant therapy with any of several agents, especially phenytoin, phenobarbital, and primidone causes disturbances in bone mineral metabolism. Anticonvulsants stimulate the hepatic microsomal mixed-oxidase enzymes and hence increase the rate of clearance of vitamin D and its metabolism. The severity of clinical manifestations in any given individual appears to be a function of the combined effects of variety of factors including drug type and total drug dose, dietary vitamin D intake, sunlight exposure, and physical activity level. We report a case of osteomalacia associated with long term carbamazepine therapy in a 21-year-old male with less exposure to sunlight(J Kor Soc Endocrinol 15:286?290, 2000).