Soon-Sun Bak

Attenuation of pro-inflammatory mediators in LPS-stimulated BV2 microglia by chitooligosaccharides via the MAPK signaling pathway

Chitooligosaccharides (COS), depolymerized products of chitosan, has received considerable attention as bioactive material due to their biocompatible, biodegradable, non-toxic and non-allergenic natures. In this study, COS of four different molecular weight ranges (<1, 1-3, 3-5 and 5-10 kDa) were investigated for their abilities to modulate inflammatory mediators in lipopolysaccharides (LPS)-stimulated BV2 microglia. At the concentration of 500 μg/ml, COS attenuate the productions of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) by inhibiting inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expressions. Furthermore, the release and expression levels of inflammatory cytokines; including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were also attenuated by COS. Notably, the inhibitory activity of COS depends significantly on its molecular weight, with lower molecular weight showed higher activity. In addition, the suppressive effects on the phosphorylation of JNK and p38 mitogen-activated protein kinase (MAPK) by COS were confirmed. These results indicate that COS could be used as an inhibitor in regulating microglial inflammatory responses. Moreover, COS may assist therapeutic treatment of neurodegenerative diseases which accompanied with microglial activation.

Anti-obesity effect of carboxymethyl chitin by AMPK and aquaporin-7 pathways in 3T3-L1 adipocytes

The aim of this study was to investigate the anti-obesity effect of carboxymethyl-chitin (CM-chitin), a water-soluble derivative of chitin, by measuring lipid accumulation and adipogenesis related factors in 3T3-L1 adipocytes. CM-chitin was synthesized by means of carboxymethylation reaction. Its inhibitory effect on lipid accumulation was investigated by measuring triglyceride content and glycerol release level. The gene and protein levels associated with adipogenesis were determined using reverse transcriptase-polymerase chain reaction and Western blot analysis. Treatment with CM-chitin reduced triglyceride content and enhanced glycerol secretion in a dose-dependent manner. CM-chitin induced the down-regulation of adipogenesis related transcriptional factors and adipocyte specific gene promoters. Moreover, the specific mechanism by CM-chitin was confirmed by transcriptional activations of the phosphorylated adenosine monophosphate-activated protein kinase (AMPK) and aquaporin-7. These results suggest that CM-chitin exerts anti-adipogenic effect on lipid accumulation through modulations of AMPK and aquaporin-7 signal pathways.

Ecklonia cava promotes hair growth

Previous studies have reported the protective effects on skin elasticity of the edible marine seaweed Ecklonia cava, which acts through regulation of both antioxidative and anti‐inflammatory responses.

[P10-206] Fermentation Properties of Young Radish Kimchi Prepared Using Young Radish Cultivated in the Soil Containing Sulfur and It's Inhibitory Effect on the Growth of AGS Human Gastric Adenocarcinoma Cells

Young radishes (YR, yeolmu in Korean) were cultivated in the soil with and without sulfur. YR-Control (without sulfur) was grown in the normal soil. YR were grown in the soil with sulfur (YR-A) and sulfur added lime mortar (YR-B) on it, respectively. Also, we prepared YR kimchis using YR-Control, YR-A and YR-B. The kimchis were fermented at for 8 weeks. The growth inhibitory effects of AGS human gastric adenocarcinoma cells of the YR samples and kimchis were investigated. YR kimchis after weeks at showed higher acidity of with pH and the YR kimchis kept approximately pH 4.0 until 8 weeks. The kimchi A and B using YR-A and YR-B showed faster fermentation time, higher level of Leuconostoc sp. and lower level of Lactobacillus sp. during the fermentation, comparing to the control kimchi using YR-Control. Juices from YR-A and YR-B showed higher growth inhibitory effects of AGS human gastric adenocarcinoma cells than the juice from YR-Control at the same concentration. The growth inhibitory effect of YR-A was similar to that of the YR-B. The kimchi A and B juices also exhibited higher inhibitory effects on the growth of AGS human gastric adenocarcinoma cells than that of the control kimchi at the higher concentration of . Methanol extracts from the YR-kimchis also led to the similar results to the results of the juices. These results suggested that preparing of kimchi using differently cultivated YR especially in the soil with sulfur, which can help to synthesize sulfur-containing compounds, could increase the growth inhibitory effects of AGS human gastric adenocarcinoma cells.

Baicalin, a flavonoid, affects the activity of human dermal papilla cells and promotes anagen induction in mice

Baicalin, a flavonoid isolated from Scutellaria baicalensis, is known to have multiple biological functions. Recent studies have demonstrated that baicalin treatment increases alkaline phosphatase activity (ALP) and osteoprotegerin secretion by osteoblasts. Furthermore, baicalin induces the differentiation of cultured osteoblasts via the activation of the Wnt/β-catenin signaling pathway. In this study, we evaluated the hair growth-promoting effects of baicalin in human follicular dermal papilla (DP) cells. A reporter assay and Western blotting were used to assess the effect of baicalin on β-catenin signaling in DP cells. ALP activity and messenger RNA (mRNA) expression were examined by ALP staining and real-time polymerase chain reaction (PCR), respectively. Growth factor expression levels were also evaluated using real-time PCR. Finally, the effect of baicalin on hair growth in vivo was examined by topical application of baicalin on the shaved dorsal skin of C57BL/6 mice. Our results indicate that baicalin activates Wnt/β-catenin signaling in a dose-dependent manner in human DP cells. ALP mRNA expression and activity were significantly induced in the presence of baicalin. In addition, treatment with baicalin induced the mRNA expression of growth factors, such as insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF). Moreover, compared to vehicle treatment, baicalin treatment induced an earlier conversion from telogen to anagen. Our results strongly suggest that baicalin promotes hair growth by regulating the activity of DP cells.

7-Phloroeckol promotes hair growth on human follicles in vitro

Abstract7-Phloroeckol, phloroglucinol derivative isolated from marine brown algae, has anti-oxidative, anti-inflammatory responses and MMP inhibitory activities. In this study, we evaluated the hair growth-promoting effects of 7-phloroeckol in human hair follicles. To investigate cell viability of human dermal papilla cells (DPCs) and outer root sheath (ORS) cells in the presence or absence of 7-phloroeckol treatment, MTT assay was employed. Moreover, gene expression and protein concentration of insulin-like growth factor (IGF)-1 was measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. 7-Phloroeckol induced an increase in proliferation of DPCs and ORS cells. In addition, hair shaft growth was measured using the hair-follicle organ culture system. 7-Phloroeckol resulted in elongation of the hair shaft in cultured human hair follicles. 7-Phloroeckol induced an IGF-1 mRNA expression and protein concentration in DPCs and conditioned media, respectively. These results suggest that 7-phloroeckol promotes hair growth through stimulation of DPCs and ORS cells.

Follistatin and secreted frizzled-related protein 1, OVO homolog-like 1-regulated genes, are important for hair follicle neogenesis

Background Recent studies showed that Wnt signalling through the b-catenin pathway maintains the hair-inducing capacity (trichogenicity) of the dermal papilla cells and that inactivation of the b-catenin gene within the dermal papilla of fully developed hair follicles prevented the normal regeneration of hair follicles (1,2). In addition, Tsai et al. (3) very recently reported that Wnt/b-catenin signalling in dermal condensates is required for hair follicle formation. In line with this, we have shown recently that significant impairment in hair follicle induction is present in b-catenin siRNA-transfected neonatal dermal cells (4). We also identified OVO homolog-like-1 (OVOL1) is one of the genes downregulated by b-catenin knockdown and found that OVOL1 siRNA-transfected neonatal dermal cells showed significantly impaired hair follicle induction (4). Furthermore, we found that hair follicle induction was significantly increased when dermal cells were transduced with lentivirus carrying OVOL1 (Lt-OVOL1), demonstrating that OVOL1 has a critical role in hair follicle neogenesis (4). Questions addressed As OVOL1 itself is a transcription factor, genes regulated by OVOL1 are thought to be involved in hair induction. In this study, as a next step, we have investigated which OVOL1-regulated genes affect the potency of hair induction in dermal cells. Experimental design To identify genes regulated by OVOL1 in neonatal mouse dermal cells, cultured neonatal dermal cells were transduced with either control lentivirus (Lt-control) or lentivirus carrying OVOL1 (LtOVOL1) for 48 h and microarray analysis were performed using the Affymetrix mouse gene ST array (Fig. 1a). To investigate whether the trichogenicity of neonatal dermal cells is affected by follistatin (Fst)or secreted frizzled-related protein 1 (SFRP1) expression, we adopted a small interfering RNA (siRNA)-mediated gene knockdown approach and a ‘patch assay’ as described previously (5), which employed three major steps: (i) isolation and cultivation of dermal and epidermal cells from mouse skin, (ii) siRNA transfection of cultured dermal cells and (iii) implantation of transfected dermal cells in combination with cultured epidermal cells. These steps are detailed in Data S1 and Fig. 2a. All animal experiments were conducted in accordance with the guidelines and following the approval of the Institutional Animal Care and Use Committee at Kyungpook National University. Results RT-PCR analysis showed that OVOL1 mRNA expression was significantly increased in Lt-OVOL1-transduced dermal cells compared with control (Fig. 1b). A number of genes were differentially regulated in Lt-OVOL1-transduced dermal cells (Tables S1 and S2). Indeed, we found OVOL1 level was 37.8-fold in Lt-OVOL1transduced dermal cells in microarray data. Some of the most upregulated and downregulated genes were validated by real-time PCR analysis (Fig. 1c). Among the OVOL1-regulated genes, we first focused on Fst because the Fst promoter is proved to contain a TCF (T-cell factor) binding site and is activated directly through the Wnt/b-catenin signalling pathway (5). We also focused on SFRP1 because SFRP1 is reported to enhance rather than antagonize Wnt signalling in some cellular contexts (6). An overall knockdown efficiency of 63% was observed in Fst siRNA-transfected dermal cells compared with control siRNAtransfected dermal cells (Fig. 2b). Co-implants of Fst siRNA-transfected dermal cells and epidermal cells showed significantly impaired hair follicle induction (Fig. 2c–e). Fst siRNA-transfected Lt-control OVOL-1