LLC-PK1 세포에서의 퓨모너신 B1에 의해 유도된 스핑고리피드 대사

유환수(Hwan Soo Yoo),윤여표(Yeo Pyo Yun) 대한약학회 1997 약학회지 Vol.41 No.6

The purpose of this study was to determine the effect of sulfinpyrazone on fumonisin B1-induced elevation of free sphingoid bases in LLC-PK1 cells. Fumonisins are a family of mycotoxins produced by the fungi Fusarium moniliforme which is common contaminant in corn. Fumonisins are also potent inhibiors of sphingosine and sphinganine N-acyltransferases (ceramide synthases), key enzymes in sphingolipid metabolism resulting in the elevation of free sphinganine. The cytosolic concentration of fumonisin B1 was known to be closely proportional to the elevation of free sphinganine in LLC-PK1 cells [Yoo, H.-S., Norred, W.P., Wang, E., Merrill, A.H., Jr., and Riley, R.T. (1992) Toxicol. Appl.Pharmacol. 114. 9-15]. Sulfinpyrazone, an anion transport inhibitor, reduced the elevated level of free sphinganine resulting from fumonisin B1 inhibition of de novo sphingolipid biosynthesis. Fumonisin B1 at a concentration of 20mcM showed approximately 120pmol/106 cells relative to 3-10pmol/106 cells in control cultures, and sulfinpyrazone at a concentration of 200mcM partially reversed the increased level of free sphinganine induced by fumonisin B1 down to normal level after exposure to fumonisin B1 for 8 to 24hr. However, the reduced effect of sulfinpyrazone on the fumonisin B1-induced elevation of intracellular sphinganine was not shown after 24hr. Fumonisin B1 exposure to LLC-PK1 cells for 36 and 48hr showed approximately 74 and 80pmol per 106 cells relative to 82 and 76pmol,respectively, in fumonisin B<1SUB>1 plus sulfinpyrazone-treated cultures. Sulfinpyrazone-induced less elevation of free sphinganine in confluent cells after exposure to fumonisin B1 suggested that either sulfinpyrazone may block the availability of fumonisin B1 to cells or act on the fumonisin B1 interaction with ceramide synthase.

실크 단백질의 식이 공급이 아토피 피부염 동물 모델 NC/Nga Mice 표피의 스핑고이드 베이스 및인산화물 함량 변화에 미치는 영향

김영애(Kim Youngae),송은화(Song Eun-hwa),신경오(Shin Kyoungoh),이용문(Lee Yongmoon),조윤희(Cho Yunhi) 韓國營養學會 2012 Journal of Nutrition and Health Vol.45 No.2

In our previous studies, dietary supplements of silk protein, sericin, and fibroin, were beneficial for improving epidermal levels of ceramides, which are the major lipids for maintaining the epidermal barrier. In this study, we investigated the dietary effects of silk protein on epidermal levels of free sphingoid bases and their phosphates such as C18 sphingosine (So), C18 sphinganine (Sa), C18 sphingosine-1-phosphate (S1P), and C18 sphinganine-1-phosphate (Sa1P), which are either synthetic substrate or degradative metabolites of ceramides. Forty-five male NC/Nga mice, an animal model of atopic dermatitis (AD), were divided into three groups: group CA was an atopic control and fed a control diet, group S was fed a 1% sericin diet, and group F was fed a 1% fibroin diet. Fifteen male BALB/c mice served as group C (control group) and were fed the control diet. All mice were fed with diets and water ad libitum for 10 weeks. Sa in group CA was lower than that in group C, but So in group CA was similar to that in group C. So and Sa were higher in groups S and F than those in group CA; So level was even higher than that in group C, and Sa level was similar to that of group C. The So/Sa ratio in group CA, which is reported to increase in AD, was significantly higher than that of group C. The So/Sa ratio was lower in groups S and F than that in group CA, and decreased further in group F. However, S1P and Sa1P in groups S and F were similar to those in group CA. Taken together, we demonstrated that silk protein, sericin and fibroin dietary supplements, increased So and Sa levels, and decreased the So/Sa ratio. (Korean J Nutr 2012; 45(2): 113 ~ 120)

Fumonisin B1-Induced Toxicity Was Not Exacerbated in Glutathione Peroxidase-1/Catalase Double Knock Out Mice

( Taddesse Yayeh ),( Ha Ram Jeong ),( Yoon Soo Park ),( Sohyeon Moon ),( Bongjun Sur ),( Hwan-soo Yoo ),( Seikwan Oh ) 한국응용약물학회 2021 Biomolecules & Therapeutics(구 응용약물학회지) Vol.29 No.1

Fumonisin B1 (FB1) structurally resembles sphingolipids and interferes with their metabolism leading to sphingolipid dysregulation. We questioned if FB1 could exacerbate liver or kidney toxicities in glutathione peroxidase 1 (Gpx1) and catalase (Cat) knockout mice. While higher serum levels of thiobarbituric acid reactive substances (TBARS) and sphinganine (Sa) were measured in Gpx1/Cat knockout mice (Gpx1/Cat KO) than wild type mice after 5 days of FB1 treatment, serum levels of alanine aminotransferase (ALT), sphingosine-1 phosphate (So-1-P), and sphinganine-1 phosphate (Sa-1-P) were found to be relatively low. Although Sa was highly elevated in Gpx1/Cat KO mice and wild mice, lower levels of So and Sa were found in both the kidney and liver tissues of Gpx/Cat KO mice than wild type mice after FB1 treatment. Paradoxically, FB1-induced cellular apoptosis and necrosis were hastened under oxidative stress in Gpx1/Cat KO mice.

Differential Effects of Fumonisin $B_1$ on Cell Death in Cultured Cells: the Significance of the Elevated Sphinganine

Yu, Chang-Hun,Lee, Yong-Moon,Yun, Yeo-Pyo,Yoo, Hwan-Soo The Pharmaceutical Society of Korea 2001 Archives of Pharmacal Research Vol.24 No.2

Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.

Altered Levels of Sphingosine and Sphinganine in Psoriatic Epidermis

문성혁,김낙인,김주영,송은화,조윤희,신민경 대한피부과학회 2013 Annals of Dermatology Vol.25 No.3

Background: Ceramides are the main lipid component of the stratum corneum and are a structurally heterogeneous and complex group of sphingolipids of which sphingoid bases are the basic structural constituents. Altered levels of sphingoid bases have been reported in skin conditions that involve dryness and barrier disruption, including atopic dermatitis. Objective: The purpose of this study was to investigate the altered levels of sphingoid bases in psoriatic epidermis and their relationship with the clinical severity of the psoriasis. Methods: Samples from the lesional and non-lesional epidermis were obtained from eight psoriasis patients. Levels of sphingosine and sphinganine were analyzed by high-performance liquid chromatography. The expression of ceramide synthase and ceramidase proteins, which are related to sphingosine and sphinganine metabolism,were measured using Western blot analysis. Results: Levels of sphingosine and sphinganine in the lesional epidermis were significantly higher than those in the non-lesional epidermis. Although there was no altered ceramide synthase and ceramidase, there was a highly significant positive correlation between the % change of ceramidase, the degradative enzyme of ceramide into sphingosine, and the Psoriasis Area Severity Index (PASI) score. Conclusion: The levels of sphingosine and sphinganine were significantly increased in psoriatic epidermis and the % change of ceramidase was positively correlated with the clinical severity of psoriasis.

Disruption of Sphingolipid Metabolism as a Potential Mechanism of Fumonisin Inhibition of Cell Growth in $LLC-PK_1$ Cells

Yoo, Hwan-Soo,Yun, Yeo-Pyo Korean Society of ToxicologyKorea Environmental Mu 1995 Toxicological Research Vol.11 No.1

Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately $333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.

Chirospecific Synthesis of D-erythro- and L-threo-Sphinganines from Sugars

Jeong, Ill-Yun,Lee, Jin-Hwan,Lee, Byong-Won,Kim, Jin-Hyo,Park, Ki-Hun Korean Chemical Society 2003 Bulletin of the Korean Chemical Society Vol.24 No.5

D-erythro-sphinganine 1 and L-threo-sphinganine 2 have been prepared in the enantiomerically pure form by the chirospecific manner. Key intermediates, 2-amino-3-hydroxy-4-pentenoates 8 and 12, were obtained from L-glucono-1,5-lactone and L-gulonic acid g-lactone via a simultaneous dealkoxyhalogenation.

Geranyllinalool에 의한 LLC-PK1 세포내 스핑고지질 생합성 억제

조양혁,이용문 대한약학회 1999 약학회지 Vol.43 No.1

Geranyllinalool, a polyisoprenoid compound, was found to block the early biosynthetic pathway of sphingolipids in LLC-PKl cells. Sphinganine, an intermediate in sphingolipid biosynthetic pathway, was abruptly accumulated in LLC-PKl cells at $2{\;}{\mu}M$ of fumonisin B1(FB1), a specific inhibitor of sphinganine N-acyltransferase, for 24 hr. Geranyllinalool lowered the $B_1(FB_1)$, a specific inhibitor of sphinganine N-acyltransferase, for 24 hr. Geranyllinalool lowered th FB1 and $50{\;}\mu$M geranyllinalool. l-Cy-closerine, an inhibitor of serine-palmitoyl transferase, was used as a positive control to evaluate the inhibitory effect of geranyllinalool. These results suggest that geranyllinalool may inhibit the serine-palmitoyl transferase, the first enzyme in de novo sphingolipid biosynthesis, resulting in the altered regulation of sphingolipid metabolism.

Stereoselective Synthesis of L-threo-Sphingosine, L-threo-Sphinganine, D-threo-Sphingosine, and D-threo-Sphinganine via Oxazoline Formation and Olefin Cross-Metathesis; Potent Protein Kinase C Inhibitor Analogues

Tian, Yong-Shou,Joo, Jae-Eun,Pham, Van-Thoai,Lee, Kee-Young,Ham, Won-Hun 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.2

In this study, we explored a convenient and concise route for synthesis of L-threo-sphingosine, D-threo-sphingosino, L-threo-sphinganine and D-threo-sphinganine from commercially available L- or D-serine. The key steps are the simple preparation of trans-oxazoline and intermolecular olefin cross metathesis.

Stereoselective Synthesis of L-threo-Sphingosine, L-threo-Sphinganine, D-threo-Sphingosine,,,,

Yong-Shou Tian,Jae-Eun Joo,Van-Thoai Pham,Kee-Young Lee,Won-Hun Ham 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.2

In this study, we explored a convenient and concise route for synthesis of L-threo-sphingosine, D-threo-sphingosine, L-threo-sphinganine and D-threo-sphinganine from commercially available L- or D-serine. The key steps are the simple preparation of trans-oxazoline and intermolecular olefin cross metathesis.