• 약학회지
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• 제43권1호
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• pp.85-90
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• 1999

FTY720, a novel immunosuppressant, elevated the level of endogenous sphingoid bases in a dose-dependent manner within 3 hr in $LLC-PK_1$ cells. The relative molar ratio of sphingoid bases expressed as sphingosine/sphinganine (SPN/SPA), a biomarker of altered sphingolipid biosynthesis, in $10{\;}{\mu}M$ of FTY720 showed tow-fold increase as compared with the one in control culture. FTY720 under the serum-free medium condition increased only cytosolic free sphingosine concentration, not sphinganine concentration in a time-dependent manner over the 8 hr incubation under the same condition as in serum free cultures, the SPN/SPA ratio began to fluctuate and the number of floating cells as an indicator of cytotoxicity was increased 8 hr after the addition of FTY720 to cultured cells. These results suggest that the process of FTY720-induced cell death in $LLC-PK_1$ cells.

• 한국식품영양과학회지
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• 제31권2호
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• pp.177-183
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• 2002

대두 및 대두발효식품인 청국장과 된장의 cerebroside 구성성분인 지방산, 당 및 장쇄 sphingoid base들을 GC-MS로 분석하였다. Cerebroside 산-가수분해물의 지방산들을 O-TMS methylester 유도체로 만들어 분석하였으며 대두 및 청국장의 경우 2-hydroxyhexadecanoic acid(16 : 0h)가 각각 52.2% 및 62.0%로 가장 많았고 2-hydroxydocosanoic acid(22 : 0h) 및 2-hydroxytetracosanoic acid(24 : 0h)도 15.0~17.6%이었다. 그러나 된장의 경우 16 : 0h는 10.2%로 많은 감소를 보인 반면 24 : 0h는 40.9%, 22 : 0h는 23.4% 었으며 23 : 0h, 25 : 0h 및 26 : 0h도 대두 및 청국장에 비하여 많았다. Cerebroside 산-가수분해물의 장쇄 sphingoid base들은 N-acetyl-O-TMS 유도체로 분석하였으며 대두 및 청국장에서 4- trans, 8-trans-sphingadienine (d18 : 2 $\Delta$$^{4trans, 8trans}$) 는 59.9% 및 44.5%이었으며 4-hydroxy-8-trans, cis-sphingening (t18 : 1$\Delta$$^{8trans or cis}$)는 20.9% 및 35.9% 이었고 된장 cerebroside 역시 dl8 : 2$\Delta$$^{4trans, 8trans}$가 가장 많았다. 그러나 phytosphingosine(t18 : 0) 및 sphingosine 이 성질체(d18 : 1)들은 거의 보이지 않았다. 또한 된장의 경우 오랜 기간 미생물들에 의하여 발효되었지만 Aspergillus 속들의 cerebroside가 가지는 것으로 알려진 2-hydroxyoxtadec-3-enoic acid(18 : 1h) 및 9-methyl-4,8-sphingadienine 등은 발견되지 않았다. 대두, 청국장 및 된장 cerebroside의 당유도체 mass spectrum 분석 결과 99%이상이 glucose 1번 탄소가 ceramide backbone에 결합된 monoglucocerebroside 인 것으로 나타났다.

• Nutrition Research and Practice
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• 제5권5호
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• pp.396-403
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• 2011

Ceramides (Cer) comprise the major constituent of sphingolipids in the epidermis and are known to play diverse roles in the outermost layers of the skin including water retention and provision of a physical barrier. In addition, they can be hydrolyzed into free sphingoid bases such as $C_{18}$ sphingosine (SO) and $C_{18}$ sphinganine (SA) or can be further metabolized to $C_{18}$ So-1-phosphate (S1P) and $C_{18}$ Sa-1-phosphate (Sa1P) in keratinocytes. The significance of ceramide metabolites emerged from studies reporting altered levels of SO and SA in skin disorders and the role of S1P and Sa1P as signaling lipids. However, the overall metabolism of sphingoid bases and their phosphates during keratinocyte differentiation remains not fully understood. Therefore, in this study, we analyzed these Cer metabolites in the process of keratinocyte differentiation. Three distinct keratinocyte differentiation stages were prepared using 0.07 mM calcium (Ca$^{2+}$) (proliferation stage), 1.2 mM Ca$^{2+}$ (early differentiation stage) in serum-free medium, or serum-containing medium with vitamin C (50 ${\mu}L$/mL) (late differentiation stage). Serum-containing medium was also used to determine whether vitamin C increases the concentrations of sphingoid bases and their phosphates. The production of sphingoid bases and their phosphates after hydrolysis by alkaline phosphatase was determined using high-performance liquid chromatography. Compared to cells treated with 0.07 mM Ca$^{2+}$, levels of SO, SA, S1P, and SA1P were not altered after treatment with 1.2 mM Ca$^{2+}$. However, in keratinocytes cultured in serum-containing medium with vitamin C, levels of SO, SA, S1P, and SA1P were dramatically higher than those in 0.07- and l.2-mM Ca$^{2+}$-treated cells; however, compared to serum-containing medium alone, vitamin C did not significantly enhance their production. Taken together, we demonstrate that late differentiation induced by vitamin C and serum was accompanied by dramatic increases in the concentration of sphingoid bases and their phosphates, although vitamin C alone had no effect on their production.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of International Convention of the Pharmaceutical Society of Korea
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• pp.287.1-287.1
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• 2001

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.162.1-162.1
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• 2003

The purpose of this study was to determine the possibility of sphingolipid as a diagnostic marker for Myocardiac Infarction(MI), atherosclerosis-related cardiovascular disease. Sphingolipids are known to playa role in the occurrence of atherosclerosis in human blood vessels. Platelet-poor plasma(PPP) and washed platelets were prepared from healthy volunteers and MI patients, and sphingolipids analyzed. (omitted)

• 한국식품영양과학회지
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• 제30권6호
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• pp.1266-1271
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• 2001

대두식품들로부터 추출한 지질분획들이 사람의 K562 백혈구암세포, mouse의 Yak1 백혈구암세포 및 S180 고형암세포에 미치는 세포독성 효과를 MTT assay법으로 조사하였다. 대두, 비지 및 된장의 총지질을 chloroform/methanol(2 : 1) 및 물포화 butanol로 추출하고 20배 용량의 acetone을 가하여 아세톤상층액(AS 획분) 및 아세톤침전물(AP 획분)을 얻었다 된장지질의 AS 획분 및 AP 획분이 실험에 사용한 K562, Yac1 및 S180암세포에 가장 강한 세포독성을 일으켰으며 비지 의 AS 및 AP 획분 역시 된장보다는 약하지만 세포독성을 나타냈으나 대두의 지질획분들은 세포독성효과를 나타내지 않았다. 된장지질의 AS 및 AP분획의 지질조성을 보면 대두 및 비지에 비하여 많은 양의 유리지방산이 함유되어 있었다. 또한 지질획 분들을 0.4 N KOH/methanol로 약알카리 분해하고 silica gel TLC로 분석한 결과, 된장의 지질획분에서는 대두 및 비지에서 보이지 않았던 알카리에 안정하고 ninhydrin에 양성인 물질들이 여러개 나타났으며 이들은 유리 sphingoid base 및 psycusin으로 추정되었다.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.144-149
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• 2001

N-acetylsphingosine (C2-ceramide) is a synthetic water-soluble ceramide mimicking the activity of natural ceramides. By fixing chiral conformation on carbon numbers 2 and 3 in the ceramide structure, four chiral C2-ceramides naming d-erythro-, I-erythro-, d-threo-and 1-three C2-ceramide were synthesized. We have investigated the chiral effects of these C2-ceramides on the sphingolipid metabolism, particularly on both the sphingolipid bio- synthetic pathway and on the degradation pathway. In both HL-60 and U937 cells, the chiral C2-ceramide ($10{\mu}\textrm{m}$) showed sphingosine accumulation monitored fluoromatrically by a high performance liquid chromatographic separation of the sphingoid bases. Most importantly, in HL-60 cells, l-erythro C2-ceramide induced a 50 fold increase in sphingosine as compared to the control, while l-threo C2-ceramide exhibited a minimal 7-fold in-crease. In contrast, sphinganine, another sphingoid base, showed less accumulation by any chiral C2-ceramide tested under the same conditions. These results suggested that chiral C2-ceramide primarilyacts on the sphingolipid degradation pathway rather than on the sphingolipid biosynthetic route. The strong $C_0/G_1$ phase arrest in the cell cycle by treatment of I-erythro C2-ceramide indicates that the blockade of the sphingolipid degradation pathway might be concomitantly involved in the dysfunction of the cell cycle. On the other hand, the fact that all chiral C2-ceramides tested failed to inhibit the activity of sphingosine kinase acting on the removal of sphingosine by producing sphingosine-1 -phosphate demonstrates that chiral C2- ceramides may increase sphingosine by activating various ceramidases by which natural ceramides are divided into sphingosine and free fatty acids. However, the precise steps involved in this interaction are still unknown.

• 분석과학
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• 제7권4호
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• pp.547-554
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• 1994

국내 자생 담자균, Phellinus ribis부터 ceramide 성분들이 존재함을 확인하고 실리카 컬럼크로마토그래피를 이용하여 분리하고 prep.-TLC로 재차 정제한 다음, 그 구조 및 조성을 검토하였다. FAB-MS 및 GC-MS를 이용하여 그 구성성분을 분석한 결과 지방산 주성분은 hydroxy fatty acid($C_{22:0}$, $C_{25:0}$, $C_{24:0}$, $C_{23:0}$)이고 sphingoid 염기는 (tri-hydroxy-sphinganine 및 동족계열의 아민으로 확인되었다.