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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).
Phospholipase D1 as a Key Enzyme for Decidualization in Human Endometrial Stromal Cells1
Yoon, Mee-Sup,Koo, Jun Bon,Jeong, Yong Geon,Kim, Yong Seok,Lee, Jung Han,Yun, Hyae Jin,Lee, Ki Sung,Han, Joong-Soo Oxford University Press 2007 BIOLOGY OF REPRODUCTION Vol.76 No.2
<P>Using primary cell cultures of human endometrial stromal cells (ES cells), we investigated the role of phospholipase D (PLD) in 8-Br-cAMP-induced decidualization, which involves morphological and biological differentiation processes. When treated with 0.5 mM 8-Br-cAMP for 12 days, ES cells were transformed into a decidualized morphology and produced significant amounts of prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Simultaneously, the activity and expression levels of PLD1 increased. In addition, removal of 8-Br-cAMP from decidualized ES cells restored the undifferentiated state, and this was accompanied by decreases in PLD1 promoter activity and PLD1 expression. Overexpression of dominant negative (DN)-PLD1 inhibited the morphological changes induced by 0.5 mM 8-Br-cAMP, whereas PLD1 overexpression induced morphological changes in the absence of 0.5 mM 8-Br-cAMP treatment. Moreover, knockdown of PLD1 by siRNA and blockage of PLD by treatment with 0.3% 1-butanol decreased PRL/IGFBP1 mRNA expression, whereas PLD1 overexpression increased PRL/IGFBP1 mRNA expression. Treatment of ES cells with phosphatidic acid (PA) for 3 days induced PRL mRNA expression and morphological changes, which implies that PA is an end-product of PLD activation-induced decidualization. In addition, pretreatment of ES cells with mepacrine decreased PRL/IGFBP1 expression and inhibited morphological change, whereas pretreatment with propranolol caused no changes, as compared to cAMP-treated cells, which suggests that PA induces decidualization through phospholipase A2 (PLA2G1B). Taken together, these results suggest that PLD1 regulates 8-Br-cAMP-induced decidualization through PLA2G1B, and that PLD1 upregulation is essential for the decidualization of ES cells.</P>
자동차 경량 대체소재로서의 Polycarbonate Grazing의 특성에 관한 연구
오미혜(Mee-Hye Oh),윤여성(Yeo-Seong Yoon),강민경(Min-Kyung Kang),이춘범(Chun-beom Lee),신재섭(Jae-Sup Shin),지영존(Young-Jon Ji) 한국자동차공학회 2005 한국자동차공학회 춘 추계 학술대회 논문집 Vol.2005 No.5_2
Polycarbonate is a highly versatile material that offers unique properties for to styling, design-driven challenges and reduces the weight compared to glass in today's highly competitive automotive market. In this study, to use polycarbonate for automobile window, we studied hard coating on polycarbonate and checked over application possibility in automobile. We formed coating layer using sol-gel process then we found the compound condition which have the superior property. We examined property of coating layer as various ratio, selectivity of solvent, aging time, etc. As a result of this sol-gel process, we have good coating layer. In this work, also we preferentially tried to manufacture automobile window.
Glutaraldehyde로 처리된 이종 및 자가 미세혈관 이식에 대한 실험적 연구
육홍미,윤진호,우동훈,김한중,김대섭 대한성형외과학회 1992 Archives of Plastic Surgery Vol.19 No.3
During microvascular procedures there frequently arises a need for a vascular graft. In this study the efficacy of preserving microvascular heterografts with glutaraldehyde tanning was investigated. These were compared with nonprepared autografts. Autogenous non-prepared rat femoral arterial grafts and glutaraldehyde-tanned rabbit femoral arterial grafts were interposed in fat femoral arteries. Ninty-one per cent and 74per cent patency rates were achieved respectively at 4weeks. This study has shown that glutaraldehyde tanning reduces the antigenicity of rabbit femoral arteries used as heterografts in the rat. This reduction in antigenicity prevented necrosis of the rabbit femoral the tendency to thrombosis, but as unacceptable occlusion rate (26% occluded) persisted. We assumed that thrombosis occurred because of incomplete reduction of the antigenicity of the heterografts. The exact mechanism resulting in occlusion of glutaraldegyde-tanned grafts is not known. Further work is therefore required to establish a tanning technique which can reduce antigenicity without leading to occlusion and without and without rendering the graft the difficult to use.