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Article ; Online: Konjac Ceramide (kCer)-Mediated Signal Transduction of the Sema3A Pathway Promotes HaCaT Keratinocyte Differentiation

Seigo Usuki / Noriko Tamura / Tomohiro Tamura / Kohei Yuyama / Daisuke Mikami / Katsuyuki Mukai / Yasuyuki Igarashi

Biology, Vol 11, Iss 121, p

2022 Volume 121

Abstract: Histamines suppress epidermal keratinocyte differentiation. Previously, we reported that konjac ceramide (kCer) suppresses histamine-stimulated cell migration of HaCaT keratinocytes. kCer specifically binds to Nrp1 and does not interact with histamine ... ...

Abstract	Histamines suppress epidermal keratinocyte differentiation. Previously, we reported that konjac ceramide (kCer) suppresses histamine-stimulated cell migration of HaCaT keratinocytes. kCer specifically binds to Nrp1 and does not interact with histamine receptors. The signaling mechanism of kCer in HaCaT cells is also controlled by an intracellular signaling cascade activated by the Sema3A-Nrp1 pathway. In the present study, we demonstrated that kCer treatment induced HaCaT keratinocyte differentiation after migration of immature cells. kCer-induced HaCaT cell differentiation was accompanied by some features of keratinocyte differentiation markers. kCer induced activating phosphorylation of p38MAPK and c-Fos, which increased the protein levels of involucrin that was the latter differentiation marker. In addition, we demonstrated that the effects of both kCer and histamines are regulated by an intracellular mechanism of Rac1 activation/RhoA inhibition downstream of the Sema3A/Nrp1 receptor and histamine/GPCR pathways. In summary, the effects of kCer on cell migration and cell differentiation are regulated by cascade crosstalk between downstream Nrp1 and histamine-GPCR pathways in HaCaT cells.
Keywords	ceramide ; konjac ; keratinocyte ; differentiation ; neuropilin1 ; Biology (General) ; QH301-705.5
Subject code	572
Language	English
Publishing date	2022-01-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: S -allyl Cysteine Enhances Testosterone Production in Mice and Mouse Testis-Derived I-10 Cells

Md Masud Rana / Kota Shiozawa / Katsuyuki Mukai / Katsuhiko Takayanagi / Koichi Eguchi / Halima Sultana / Yusuke Ohsaki / Michio Komai / Hitoshi Shirakawa

Molecules, Vol 26, Iss 1697, p

2021 Volume 1697

Abstract: Hypogonadism, associated with low levels of testosterone synthesis, has been implicated in several diseases. Recently, the quest for natural alternatives to prevent and treat hypogonadism has gained increasing research interest. To this end, the present ... ...

Abstract	Hypogonadism, associated with low levels of testosterone synthesis, has been implicated in several diseases. Recently, the quest for natural alternatives to prevent and treat hypogonadism has gained increasing research interest. To this end, the present study explored the effect of S -allyl cysteine (SAC), a characteristic organosulfur compound in aged-garlic extract, on testosterone production. SAC was administered at 50 mg/kg body weight intraperitoneally into 7-week-old BALB/c male mice in a single-dose experiment. Plasma levels of testosterone and luteinizing hormone (LH) and testis levels of proteins involved in steroidogenesis were measured by enzymatic immunoassay and Western blot, respectively. In addition, mouse testis-derived I-10 cells were also used to investigate the effect of SAC on steroidogenesis. In the animal experiment, SAC significantly elevated testosterone levels in both the plasma and the testis without changing the LH level in plasma and increased phosphorylated protein kinase A (p-PKA) levels. Similar results were also observed in I-10 cells. The findings demonstrating the increasing effect of SAC on p-PKA and mRNA levels of Cyp11a suggest that SAC increases the testosterone level by activating the PKA pathway and could be a potential target for hypogonadism therapeutics.
Keywords	S -allyl cysteine ; testosterone ; protein kinase A ; Organic chemistry ; QD241-441
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Neurite Outgrowth and Morphological Changes Induced by 8-trans Unsaturation of Sphingadienine in kCer Molecular Species

Seigo Usuki / Noriko Tamura / Tomohiro Tamura / Kunikazu Tanji / Daisuke Mikami / Katsuyuki Mukai / Yasuyuki Igarashi

International Journal of Molecular Sciences, Vol 20, Iss 9, p

2019 Volume 2116

Abstract: Konjac ceramide (kCer), which consists of plant-type molecular species of characteristic shingoid bases and fatty acids, is prepared from konjac glucosylceramide GlcCer by chemoenzymatical deglucosylation. kCer activates the semaphorin 3A (Sema3A) ... ...

Abstract	Konjac ceramide (kCer), which consists of plant-type molecular species of characteristic shingoid bases and fatty acids, is prepared from konjac glucosylceramide GlcCer by chemoenzymatical deglucosylation. kCer activates the semaphorin 3A (Sema3A) signaling pathway, inducing collapsin response mediator protein 2 (CRMP2) phosphorylation. This results in neurite outgrowth inhibition and morphological changes in remaining long neurites in PC12 cells. Whether a specific molecular species of kCer can bind to the Sema3A receptor (Neuropilin1, Nrp1) and activate the Sema3A signaling pathway remains unknown. Here, we prepared kCer molecular species using endoglycoceramidase I-mediated deglucosylation and examined neurite outgrowth and phosphorylation of collapsin response mediator protein 2 in nerve growth factor (NGF)-primed cells. The 8-trans unsaturation of sphingadienine of kCer was essential for Sema3A-like signaling pathway activation. Conversely, 8-cis unsaturation of kCer molecular species had no effect on Sema3A-like activation, and neurite outgrowth inhibition resulted in remaining short neurites. In addition, α-hydroxylation of fatty acids was not associated with the Sema3A-like activity of the kCer molecular species. These results suggest that 8-trans or 8-cis isomerization of sphingadienine determines the specific interactions at the ligand-binding site of Nrp1.
Keywords	ceramide ; konjac ; semaphorin3A ; neurite outgrowth ; neuropilin1 ; endoglycoceramidase ; sphingadienine ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Language	English
Publishing date	2019-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Blood-brain barrier permeability analysis of plant ceramides.

Koichi Eguchi / Daisuke Mikami / Hui Sun / Takuya Tsumita / Kaori Takahashi / Katsuyuki Mukai / Kohei Yuyama / Yasuyuki Igarashi

PLoS ONE, Vol 15, Iss 11, p e

2020 Volume 0241640

Abstract: Ceramides, a type of sphingolipid, are cell membrane components and lipid mediators that modulate a variety of cell functions. In plants, ceramides are mostly present in a glucosylated glucosylceramide (GlcCer) form. We previously showed that oral ... ...

Abstract	Ceramides, a type of sphingolipid, are cell membrane components and lipid mediators that modulate a variety of cell functions. In plants, ceramides are mostly present in a glucosylated glucosylceramide (GlcCer) form. We previously showed that oral administration of konjac-derived GlcCer to a mouse model of Alzheimer's disease reduced brain amyloid-β and amyloid plaques. Dietary plant GlcCer compounds are absorbed as ceramides, but it is unclear whether they can cross the blood-brain barrier (BBB). Herein, we evaluated the BBB permeability of synthetic plant-type ceramides (4, 8-sphingadienine, d18:2) using mouse and BBB cell culture models, and found that they could permeate the BBB both in vivo and in vitro. In addition, administrated ceramides were partially metabolized to other sphingolipid species, namely sphingomyelin (SM) and GlcCer, while crossing the BBB. Thus, plant ceramides can cross the BBB, suggesting that ceramides and their metabolites might affect brain functions.
Keywords	Medicine ; R ; Science ; Q
Subject code	580
Language	English
Publishing date	2020-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Suppression of Sebum Production and Accumulation by β -Cryptoxanthin Due to the Inhibition of the Expression of Diacylglycerol Acyltransferase-1 and Perilipin in Hamster Sebocytes

Akira Ito / Yoshiyuki Shirakura / Katsuyuki Mukai / Takashi Sato

Journal of Cosmetics, Dermatological Sciences and Applications, Vol 03, Iss 01, Pp 99-

2013 Volume 106

Abstract: Background : Acne vulgaris is characterized by the enhancement of sebaceous lipogenesis and sebum secretion, and apart from retinoids and some natural products there are few effective antiacne agents that directly suppress sebum production and ... ...

Abstract	Background : Acne vulgaris is characterized by the enhancement of sebaceous lipogenesis and sebum secretion, and apart from retinoids and some natural products there are few effective antiacne agents that directly suppress sebum production and accumulation in sebaceous glands. Objective : We examined the effects of β -cryptoxanthin ( β -CRX), which is a carotenoid pigment most abundant in Citrus unshiu Marcovich (Satsuma mandarin orange) and plays a role as a vitamin A precursor on sebum production and accumulation in hamster sebaceous gland cells (sebocytes). Materials and methods : The regulation of sebum production was examined by the measurement of triacylglycerols (TGs), the major sebum component, and oil red O staining in insulindifferentiated hamster sebocytes. The expression of diacylglycerol acyltransferase-1 (DGAT-1), a rate-limiting enzyme of TG biosynthesis, and perilipin 1 (PLIN1), a lipid storage droplet protein, was analyzed using real-time PCR and Western blotting. Results : Hamster sebocytes constitutively produced TGs during cultivation and the production of TGs was enhanced by insulin treatment. Both constitutive and insulin-enhanced TG productions were dose- and time-dependently inhibited by β -CRX as well as 13- cis retinoic acid. In addition, the gene expression of DGAT-1 was suppressed by β -CRX in the sebocytes. Furthermore, the insulin-en - hanced sebum accumulation as lipid droplets was reduced in the β -CRX-treated cells. Moreover, β -CRX was found to suppress the gene expression and production of PLIN 1 in insulin-differentiated hamster sebocytes. Conclusions : These results provide novel evidence that β -CRX is an effective candidate for acne therapy by its ability to exert dual inhibitory actions against DGAT-1-dependent TG production and PLIN1-mediated lipiddroplet formation in hamster sebocytes.
Keywords	β -Cryptoxanthin ; Sebocytes ; Triacylglycerol Biosynthesis ; Diacyglycerol Acyltransferase ; Perilipin ; Lipid-Droplet Formation ; Sebum ; Chemistry ; QD1-999 ; Science ; Q ; DOAJ:Chemistry (General) ; DOAJ:Chemistry
Subject code	620
Language	English
Publishing date	2013-03-01T00:00:00Z
Publisher	Scientific Research Publishing
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Plant sphingolipids promote extracellular vesicle release and alleviate amyloid-β pathologies in a mouse model of Alzheimer’s disease

Kohei Yuyama / Kaori Takahashi / Seigo Usuki / Daisuke Mikami / Hui Sun / Hisatoshi Hanamatsu / Junichi Furukawa / Katsuyuki Mukai / Yasuyuki Igarashi

Scientific Reports, Vol 9, Iss 1, Pp 1-

2019 Volume 11

Abstract: Abstract The accumulation of amyloid-β protein (Aβ) in brain is linked to the early pathogenesis of Alzheimer’s disease (AD). We previously reported that neuron-derived exosomes promote Aβ clearance in the brains of amyloid precursor protein transgenic ... ...

Abstract	Abstract The accumulation of amyloid-β protein (Aβ) in brain is linked to the early pathogenesis of Alzheimer’s disease (AD). We previously reported that neuron-derived exosomes promote Aβ clearance in the brains of amyloid precursor protein transgenic mice and that exosome production is modulated by ceramide metabolism. Here, we demonstrate that plant ceramides derived from Amorphophallus konjac, as well as animal-derived ceramides, enhanced production of extracellular vesicles (EVs) in neuronal cultures. Oral administration of plant glucosylceramide (GlcCer) to APP overexpressing mice markedly reduced Aβ levels and plaque burdens and improved cognition in a Y-maze learning task. Moreover, there were substantial increases in the neuronal marker NCAM-1, L1CAM, and Aβ in EVs isolated from serum and brain tissues of the GlcCer-treated AD model mice. Our data showing that plant ceramides prevent Aβ accumulation by promoting EVs-dependent Aβ clearance in vitro and in vivo provide evidence for a protective role of plant ceramides in AD. Plant ceramides might thus be used as functional food materials to ameliorate AD pathology.
Keywords	Medicine ; R ; Science ; Q
Subject code	580
Language	English
Publishing date	2019-11-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Konjac ceramide (kCer) regulates keratinocyte migration by Sema3A-like repulsion mechanism

Seigo Usuki / Noriko Tamura / Tomohiro Tamura / Shigeki Higashiyama / Kunikazu Tanji / Susumu Mitsutake / Asuka Inoue / Junken Aoki / Katsuyuki Mukai / Yasuyuki Igarashi

Biochemistry and Biophysics Reports, Vol 17, Iss , Pp 132-

2019 Volume 138

Abstract: Previously, we proposed the following mechanism for konjac ceramide (kCer)-mediated neurite outgrowth inhibition: kCer binds to Nrp as a Sema3A agonist, resulting in Nrp1/PlexA complex formation and activation of the Sema3A signaling pathway to induce ... ...

Abstract	Previously, we proposed the following mechanism for konjac ceramide (kCer)-mediated neurite outgrowth inhibition: kCer binds to Nrp as a Sema3A agonist, resulting in Nrp1/PlexA complex formation and activation of the Sema3A signaling pathway to induce phosphorylation of CRMP2 and microtubule depolymerization. The Sema3A/Nrp1 signaling pathway is known to be also expressed in normal human keratinocytes. To determine whether kCer can function in human keratinocytes as it does in neurites, that is, if it can bind to Nrp1 in place of Sema3A, we studied the effect of kCer on HaCaT cell migration activity. Using a trans-well chamber assay, we compared the effects of Sema3A and kCer on serum-derived cell migration activity. kCer showed Sema3A-like suppression of cell migration activity and induction of cellular Cofilin phosphorylation. In addition, kCer and Sema3A inhibited histamine (His)-enhanced migration of immature HaCaT cells. We have demonstrated that kCer does not interact with histaime receptors H1R or H4R directly, but we speculate that kCer may transduce a signal downstream of the His signaling pathway.
Keywords	Biology (General) ; QH301-705.5 ; Biochemistry ; QD415-436
Subject code	572
Language	English
Publishing date	2019-03-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Chemoenzymatically prepared konjac ceramide inhibits NGF-induced neurite outgrowth by a semaphorin 3A-like action

Seigo Usuki / Noriko Tamura / Shota Sakai / Tomohiro Tamura / Katsuyuki Mukai / Yasuyuki Igarashi

Biochemistry and Biophysics Reports, Vol 5, Iss C, Pp 160-

2016 Volume 167

Abstract: Dietary sphingolipids such as glucosylceramide (GlcCer) are potential nutritional factors associated with prevention of metabolic syndrome. Our current understanding is that dietary GlcCer is degraded to ceramide and further metabolized to sphingoid ... ...

Abstract	Dietary sphingolipids such as glucosylceramide (GlcCer) are potential nutritional factors associated with prevention of metabolic syndrome. Our current understanding is that dietary GlcCer is degraded to ceramide and further metabolized to sphingoid bases in the intestine. However, ceramide is only found in trace amounts in food plants and thus is frequently taken as GlcCer in a health supplement. In the present study, we successfully prepared konjac ceramide (kCer) using endoglycoceramidase I (EGCase I). Konjac, a plant tuber, is an enriched source of GlcCer (kGlcCer), and has been commercialized as a dietary supplement to improve dry skin and itching that are caused by a deficiency of epidermal ceramide. Nerve growth factor (NGF) produced by skin cells is one of the itch factors in the stratum corneum of the skin. Semaphorin 3A (Sema 3A) has been known to inhibit NGF-induced neurite outgrowth of epidermal nerve fibers. It is well known that the itch sensation is regulated by the balance between NGF and Sema 3A. In the present study, while kGlcCer did not show an in vitro inhibitory effect on NGF-induced neurite outgrowth of PC12 cells, kCer was demonstrated to inhibit a remarkable neurite outgrowth. In addition, the effect of kCer was similar to that of Sema 3A in cell morphological changes and neurite retractions, but different from C2-Ceramide. kCer showed a Sema 3A-like action, causing CRMP2 phosphorylation, which results in a collapse of neurite growth cones. Thus, it is expected that kCer is an advanced konjac ceramide material that may have neurite outgrowth-specific action to relieve uncontrolled and serious itching, in particular, from atopic eczema.
Keywords	Ceramide ; Konjac ; NGF ; Semaphorin 3A ; Neurite outgrowth ; CRMP2 ; Biology (General) ; QH301-705.5 ; Biochemistry ; QD415-436
Subject code	571
Language	English
Publishing date	2016-03-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Nrp1 is Activated by Konjac Ceramide Binding-Induced Structural Rigidification of the a1a2 Domain

Seigo Usuki / Yoshiaki Yasutake / Noriko Tamura / Tomohiro Tamura / Kunikazu Tanji / Takashi Saitoh / Yuta Murai / Daisuke Mikami / Kohei Yuyama / Kenji Monde / Katsuyuki Mukai / Yasuyuki Igarashi

Cells, Vol 9, Iss 2, p

2020 Volume 517

Abstract: Konjac ceramide (kCer) is a plant-type ceramide composed of various long-chain bases and α-hydroxyl fatty acids. The presence of d4t,8t-sphingadienine is essential for semaphorin 3A (Sema3A)-like activity. Herein, we examined the three neuropilin 1 (Nrp1) ...

Abstract	Konjac ceramide (kCer) is a plant-type ceramide composed of various long-chain bases and α-hydroxyl fatty acids. The presence of d4t,8t-sphingadienine is essential for semaphorin 3A (Sema3A)-like activity. Herein, we examined the three neuropilin 1 (Nrp1) domains (a1a2, b1b2, or c), and found that a1a2 binds to d4t,8t-kCer and possesses Sema3A-like activity. kCer binds to Nrp1 with a weak affinity of μM dissociation constant (Kd). We wondered whether bovine serum albumin could influence the ligand−receptor interaction that a1a2 has with a single high affinity binding site for kCer (Kd in nM range). In the present study we demonstrated the influence of bovine serum albumin. Thermal denaturation indicates that the a1a2 domain may include intrinsically disordered region (IDR)-like flexibility. A potential interaction site on the a1 module was explored by molecular docking, which revealed a possible Nrp1 activation mechanism, in which kCer binds to Site A close to the Sema3A-binding region of the a1a2 domain. The a1 module then accesses a2 as the IDR-like flexibility becomes ordered via kCer-induced protein rigidity of a1a2. This induces intramolecular interaction between a1 and a2 through a slight change in protein secondary structure.
Keywords	ceramide ; konjac ; semaphorin3a ; neurite outgrowth ; neuropilin 1 ; endoglycoceramidase ; sphingadienine ; Biology (General) ; QH301-705.5
Subject code	500
Language	English
Publishing date	2020-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Effects of β-Cryptoxanthin on Bone Metabolism in a Rat Model of Osteoporosis

Seiji Kusuhara / Katsuyuki Mukai / Toshiko Suzuki / Toshie Sugiyama / Noriko Ikeda

Journal of Animal and Veterinary Advances, Vol 11, Iss 1, Pp 30-

2012 Volume 35

Abstract: Osteoporosis is a bone disease in which Bone Mineral Density (BMD) is reduced with a consequent increase in the risk of bone fractures. β-cryptoxanthin (β-CRP) is present in large amounts in Satsuma mandarins and was recently reported to stimulate bone ... ...

Abstract	Osteoporosis is a bone disease in which Bone Mineral Density (BMD) is reduced with a consequent increase in the risk of bone fractures. β-cryptoxanthin (β-CRP) is present in large amounts in Satsuma mandarins and was recently reported to stimulate bone formation. In this study, researchers investigated the effects of β-CRP in Satsuma Mandarin Pulp (SMP; 2 mg g -1 β-CRP) on bone metabolism in an Ovariectomized (OVX) rat model of osteoporosis. Female rats (12 weeks of age) were ovariectomized and orally administered vehicle, 0.03 g day -1 SMP or 0.3 g day -1 SMP for 5 weeks. After that serum concentrations of osteocalcin (an osteoblastic bone formation marker) tended to be higher in the SMP groups than in the OVX vehicle group while those of collagen type I degradation products (an osteoclastic bone resorption marker) tended to be lower in the SMP groups. By bone histomorphometry, bone trabecular volume/tissue volume ratios and trabecular numbers were significantly higher in the SMP groups than in the OVX vehicle group while trabecular separation and osteoclast number/bone surface ratios were significantly lower in the SMP groups. By immunohistochemistry, percentage areas of osteocalcin immunoreactivity on trabecular surface were significantly greater in the SMP groups than in the OVX vehicle group. Dual-energy X-ray absorptiometry analyses revealed that BMDs of the lumbar vertebrae and femora, tibiae tended to increase as the dose of SMP increased in the OVX rats. In conclusion, oral SMP administration stimulated osteoblastic bone formation and inhibited osteoclastic bone resorption in OVX rats, thereby preventing the bone loss associated with osteoporosis.
Keywords	Bone metabolism ; B-cryptoxanthin ; osteoporosis ; rat ; Satsuma mandarin ; Animal culture ; SF1-1100 ; Agriculture ; S ; DOAJ:Animal Sciences ; DOAJ:Agriculture and Food Sciences
Subject code	616
Language	English
Publishing date	2012-01-01T00:00:00Z
Publisher	Medwell Online
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