• Journal of Microbiology and Biotechnology
• /
• v.25 no.1
• /
• pp.119-126
• /
• 2015

Among the various human growth factors, epidermal growth factor (hEGF, consisting of 53 amino acids) has various effects on cell regeneration, stimulation of proliferation, migration of keratinocytes, formation of granulation tissues, and stimulation of fibroblast motility, which are important for wound healing. Owing to their multiple activities, EGFs are used as pharmaceutical and cosmetic agents. However, their low productivity, limited target specificity, and short half-life inhibit their application as therapeutic agents. To overcome these obstacles, we fused the collagen-binding domain (CBD) of Vibrio mimicus metalloprotease to EGF protein. About 18 or 12 amino acids (aa) (of the 33 total amino acids), which were essential for collagen-binding activity, were combined with the N- and C-termini of EGF. We constructed, expressed, and purified EGF (53 aa)-CBD (18 aa), EGF (53 aa)-CBD (12 aa), CBD (18 aa)-EGF (53 aa), and CBD (12 aa)-EGF (53 aa). These purified recombinant proteins increased the numbers of cells in treated specimens compared with non-treated specimens and control hEGF samples. The collagen-binding activities were also evaluated. Furthermore, CBD-hybridized hEGF induced phosphorylation of the EGF receptor. These results suggested that these fusion proteins could be applicable as small therapeutic agents in wound tissue healing.

• Journal of Periodontal and Implant Science
• /
• v.34 no.1
• /
• pp.205-221
• /
• 2004

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.20 no.1 s.32
• /
• pp.27-37
• /
• 2007

Background and Objective : Psoriasis is a chronic skin disease characterized by angiogenesis. It has been reported that growth factor as vascular endothelial growth factor (VEGF) and insulin-like growth factor (IGF)-II are overexpressed in psoriatic epidermis. This stydy was carried out for whether SB extracts have an anti-angiogenic effect for angiogenic factor. Method : To investigate the inhibitory effect of VEGF expression by the SB extracts, we performed MTS assay, western blots using HaCaT cells. HaCaT cells were pretreated with SB extracts for 1 hour followed by treatment with IGF-II. Result : SB extracts significantly reduced IGF-II induced HIF-1 ${\alpha}$ protein level via p53 and MAPK pathway in HaCaT cells. Also, SB extracts inhibited IGF-II induced VEGF mRNA and protein expression levels in the HaCaT keratinocytes. Conclusion : These results suggest that inhibition of HIF-1 ${\alpha}$ and VEGF expressions by SB extracts contributes to the anti-angiogenic effects.

• Korean Journal of Acupuncture
• /
• v.26 no.2
• /
• pp.27-38
• /
• 2009

Objectives: Rheumatoid arthritis (RA) is a chronic autoimmune disease, principally characterized by synovial inflammation of the joints. We previously reported the effect of acupuncture for RA, but the mechanism is still unclear. Various factors such as oxidative stress and angiogenesis were involved in the pathogenesis of RA, and recently, it has also been reported that cytokines also play a major role in RA. Thus, we investigated whether acupuncture could induce any changes in the levels of cytokines including vascular endothelial growth factor (VEGF), angiogenin and epidermal growth factor (EGF) as well as erythrocyte sedimentation rate (ESR), c-reactive protein (CRP), and rheumatoid factor (RF) in the sera of RA patients. Methods: The forty three patients who met the American College of Rheumatology (ACR) criteria for RA recruited. The acupuncture group (n=21) underwent 14 sessions of partially individualized acupuncture treatment for 6 weeks, and the control group had no treatment (n=13) over the same periods. We evaluated ESR, CRP and RF. In addition, to find out the mechanism of acupuncture, we assessed the changes of the cytokine activities using protein cytokine array in the sera of the patients. Results: Acupuncture significantly decreased the levels of ESR and CRP, but RF were not changed after 6-week acupuncture treatments. Moreover, acupuncture reduced the levels of VEGF, angiogenin and EGF in the sera of the patients. Interestingly, they were related with angiogenesis, which is an important process in the pathogenesis of RA. The levels of oncostatin, interleukin(IL)-$1{\alpha}$, IL-8, leptin, monocyte chemotactic protein-1, macrophage-derived chemokine, macrophage inflammatory proteins-1, platelet-derived growth factor BB and RANTES were not changed significantly. Conclusions: The effect of acupuncture for reliving RA symptoms can be partially explained by inhibition of angiogenesis factors in the sera of the RA patients.

• Biomolecules & Therapeutics
• /
• v.7 no.4
• /
• pp.329-334
• /
• 1999

Recently, cellular signal transduction mechanisms are greatly understood. However, bone cell signaling is not completely characterized. Interestingly, bone cells synthesize a number of growth factors such as IGF-I PDGF, IGF-II etc., suggesting these growth factors play important roles in bone cell signaling. In the present study, potential roles of nitric oxide (NO) and protein kinases in osteoblast signal transduction are proposed.

• Journal of Aquaculture
• /
• v.21 no.4
• /
• pp.239-243
• /
• 2008

A $3{\times}2$ factorial experiment was conducted to investigate the proper dietary protein and lipid levels for the growth of sub-adult flounder Paralichthys olivaceus reared during the summer season. Six experimental diets were formulated to contain three levels of protein (45%, 50% and 55%) and two levels of lipid (9% and 14%). Duplicate groups of fish (initial body weight of 298 g) were hand-fed to apparent satiation during the summer season ($21.8{\pm}1.7^{\circ}C$) for 60 days. Survival of each group was over 83% and there was not significant difference among all groups. Weight gain of fish fed the 45% protein diet with 14% lipid was not significantly different from that of fish fed the 50% and 55% protein diets with 9% and 14% lipids, but weight gain of fish fed the 45% protein diet with 9% lipid was significantly lower than that of fish fed the 55% protein diets with 14% lipid. Feed efficiency tended to increase with increasing dietary lipid level at each protein levels, although no significant differences were observed at 50% and 55% protein levels. Protein efficiency ratio, daily feed intake, condition factor and hepatosomatic index were not significantly affected by dietary protein and lipid levels. Crude lipid content of the liver tended to increase with increasing dietary lipid level at the same protein levels, but the opposite appearance was found for moisture content. The contents of moisture, crude protein and crude lipid of the dorsal muscle were not significantly affected by dietary protein and lipid levels. Based on data obtained form this study, inclusion of dietary protein at level of 45% appears sufficient to support optimal growth, and an increase of dietary lipid level from 9% to 14% has beneficial effects on feed utilization of sub-adult flounder during the summer season.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.1
• /
• pp.3-7
• /
• 2005

The insulin-like growth factors (IGFs), their receptors, and their binding proteins play key roles in regulating cell proliferation and apoptosis. Insulin-like growth factor binding protein-3 (IGFBP3, OMIM #146732) is one of the proteins that bind to the IGFs. IGFBP3 is a modulator of IGF bioactivity, and direct growth inhibitor in the extravascular tissue compartment. We identified twenty-two novel single nucleotide polymorphisms (SNPs) in IGFBP3 gene in Korean cattle (Hanwoo, Bos taurus coreanae) by direct sequencing of full gene including -1,500 bp promoter region. Among the identified SNPs, five common SNPs were screened in 650 Korean cattle; one SNP in promoter (IGFBP3 G-854C), one in 5'UTR region (IGFBP3 G-100A), two in intron 1 (IGFBP3 G+421T, IGFBP3 T+1636A), and one in intron 2 (IGFBP3 C+3863A). The frequencies of each SNP were 0.357 (IGFBP3 G-854C), 0.472 (IGFBP3 G-100A), 0.418 (IGFBP3 G+421T), 0.363 (IGFBP3 T+1636A) and 0.226 (IGFBP3 C+3863A), respectively. Haplotypes and their frequencies were estimated by EM algorithm. Six haplotypes were constructed with five SNPs and linkage disequilibrium coefficients (|D'|) between SNP pairs were also calculated. The information on SNPs and haplotypes in IGFBP3 gene could be useful for genetic studies of this gene.

• The Korean Journal of Physiology and Pharmacology
• /
• v.26 no.6
• /
• pp.469-478
• /
• 2022

WNT signaling plays an important role in cardiac development, but abnormal activity is often associated with cardiac hypertrophy, myocardial infarction, remodeling, and heart failure. The effect of WNT signaling on regulation of atrial natriuretic peptide (ANP) secretion is unclear. Therefore, the purpose of this study was to investigate the effect of Wnt agonist 1 (Wnta1) on ANP secretion and mechanical dynamics in beating rat atria. Wnta1 treatment significantly increased atrial ANP secretion and pulse pressure; these effects were blocked by U73122, an antagonist of phospholipase C. U73122 also abolished the effects of Wnta1-mediated upregulation of protein kinase C (PKC) β and γ expression, and the PKC antagonist Go 6983 eliminated Wnta1-induced secretion of ANP. In addition, Wnta1 upregulated levels of phospho-transforming growth factor-β activated kinase 1 (p-TAK1), TAK1 banding 1 (TAB1) and phospho-activating transcription factor 2 (p-ATF2); these effects were blocked by both U73122 and Go 6983. Wnta1-induced ATF2 was abrogated by inhibition of TAK1. Furthermore, Wnta1 upregulated the expression of T cell factor (TCF) 3, TCF4, and lymphoid enhancer factor 1 (LEF1), and these effects were blocked by U73122 and Go 6983. Tak1 inhibition abolished the Wnta1-induced expression of TCF3, TCF4, and LEF1 and Wnta1-mediated ANP secretion and changes in mechanical dynamics. These results suggest that Wnta1 increased the secretion of ANP and mechanical dynamics in beating rat atria by activation of PKC-TAK1-ATF2-TCF3/LEF1 and TCF4/LEF1 signaling mainly via the WNT/Ca²⁺ pathway. It is also suggested that WNT-ANP signaling is implicated in cardiac physiology and pathophysiology.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.28 no.1
• /
• pp.178-190
• /
• 1999

This study was conducted to develop a simulation model for the growth dynamics of pigs and to describe quantitatively protein deposition depending on the amino acid composition of feed protein. In the model it is assumed that the essential processes that determine the utilization of feed protein in the whole body are protein synthesis, breakdown of protein, and oxidation of amino acid. Besides, it is also assumed that occurrence of protein deposition depends on genetic potential and amino acid composition of feed protein. The genetic potential for the protein deposition is the maximum capacity of protein synthesis, being dependent on the protein mass of the whole body. To describe the effect of amino acid composition of feed on the protein deposition, a factor, which consist of ten amino acid functions and lie between 0 and 1, is introduced. Accordingly a model was developed, which is described with 15 flux equations and 11 differential equations and is composed of two compartments. The model describes non linear structure of the protein utilization system of an organism, which is in non steady state. The objective function for the simulation was protein deposition(g/day) cal culated according to the empirical model, PAF(product of amino acid functions) of Menke. The mean of relative difference between the simulated protein deposition and PAF calculated values, lied in a range of 11.8%. The simulated protein synthesis and breakdown rates(g/day) in the whole body showed a parallel behavior in the course of growth.

• Proceedings of the Korean Nutrition Society Conference
• /
• 1999.05b
• /
• pp.39-40
• /
• 1999