• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.1
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• pp.11-24
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• 2022

It has been reported that the ethanolic extract of the root of Piper methysticum (P. methysticum) inhibits melanogenesis in melanocyte stimulating hormone (MSH)-activated B16 melanoma cells. Flavokawain B (FKB) and Flavokawain C (FKC) isolated from this extract have been found to inhibit melanin production based on anti-melanogenesis activity. This study was designed to find out the inhibition and its process of FKB and FKC on melanin synthesis in melan-a melanocytes. FKB and FKC inhibited melanogenesis at 10 μM, 5 μM respectively in melan-a melanocytes. However, they did not inhibit extracellular tyrosinase activity from melan-a melanocytes. FKB reduced the protein level of tyrosinase (Tyr), tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF) and the mRNA level of Tyr and TRP-1. FKC reduced the protein level of TRP-2 and MITF and the mRNA level of TRP-1 and Tyr. The reduced expression of Tyr and TRP-1 might be resulted from the decreased MITF which regulates major melanogenic proteins. However, since the mRNA expression of MITF did not change by FKB and FKC treatment, the effects of FKB and FKC on extracellular signal regulating kinase (ERK)/AKT phosphorylation, known to regulate the degradation of MITF, were confirmed. FKB and FKC significantly increased the phosphorylation of ERK1/2, not in AKT. These results suggest that FKB and FKC may be helpful as a potential depigmenting agent for various hyper-pigmentary disorders.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.3
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• pp.237-245
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• 2011

In order to investigate the possibility of Inonotus obliquus extract as an active ingredient for wrinkle-care cosmetics, we prepared 70 % ethanolic extract of Inonotus obliquus and measured its DPPH radical scavenging activity and elastase inhibitory activity. We also evaluated the effect of Inonotus obliquus extract on expression of MMPs and HAS-2 in fibroblast and HaCaT cell, respectively. Inonotus Obliquus extract showed DPPH radical scavenging activity and elastase inhibitory activity in a dose-dependant manner ($SC_{50}$ = 91 ${\mu}g$/mL, $IC_{50}$=124 ${\mu}g$/mL). Inonotus obliquus extract inhibited the expression of MMP-1 mRNA 50 ~ 79 % at concentrations of 5 ~ 25 ${\mu}g$/mL, and reduced the expression of MMP-2 around 20 % at a concentration of 10 ${\mu}g$/mL. And it also reduced the expression of MMP-9 54 ~ 70 % in tconcentrations of 5 to 25 ${\mu}g$/mL. Furthermore, Inonotus obliquus extract increased HAS-2 mRNA expression in a dose-dependent manner in concentrations of 5 to 25 ${\mu}g$/mL without cytotoxicity. These results suggested that Inonotus Obliquus extract could be useful as an active ingredient for wrinkle-care cosmetics.

• The Journal of the Korea Contents Association
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• v.19 no.11
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• pp.375-382
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• 2019

Abnormalities of trophoblast due to early inflammation in pregnancy increase the expression of CRP and affect maternal-fetal interactions, leading to preterm birth and preeclampsia. However, biomarkers related to the regulation of CRP expression have not been found. In this study, miRNA associated with increased expression of CRP was identified and their expression was analyzed to reveal biomarkers involved in the regulation mechanism of trophoblast inflammation through miRNAs. miRNAs that were predicted to regulate CRP gene expression in miRNA databases (mirna, TargetScan, MicroCosm) were screened and HTR-8/SVneo cell lines were treated with LPS (20 ng/mL) to induce inflammatory responses in vitro, with selected miR-7, miR-150, miR-186 and miR-424. The expression was analyzed by qRT-PCR. As a result, expression of CRP was significantly increased in LPS-treated trophoblast (p<0.001) and miR-150 and miR-424 expression were significantly decreased (p<0.001). Thus, miR-150 and miR-424 are involved in the regulation of CRP expression in inflammatory-induced trophoblast and may be useful for the prenatal diagnosis of inflammatory obstetric diseases.

• Development and Reproduction
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• v.15 no.4
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• pp.349-357
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• 2011

Since nesfatin-1/NUCB2 involved in the control of appetite and energy metabolism was discovered for the first time in hypothalamus, many reports have shown its expression in various tissues. We also recently demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs of mouse. However, no data exist on nesfatin-1/NUCB2 expression, regulation, and secretion in the uterus. Therefore, we examined the expression of nesfatin-1/NUCB2 in mouse uterus and the effects of PMSG and estrogen on its expression. NUCB2 mRNA expression in the uterus was determined by conventional and real-time PCR and nesfatin-1 protein expression was detected by western blotting. In immunohistochemistry staining, nesfatin-1 protein was localized at the epithelial cells of the uterine glands and endometrium. Nesfatin-1 protein binding sites were displayed at the epithelial cells of uterine glands and specific granulocytes including neutrophils. Additionally, to examine if the nesfatin-1/NUCB2 expression in the uterus is regulated by gonadotropin or estrogen, ovariectomized mice were treated with PMSG or $17{\beta}$-estradiol. The expression levels of NUCB2 mRNA in the uterus was significantly increased in the control mice after PMSG treatment, but not in the ovariectomized mice. In contrast, NUCB2 mRNA expression was dramatically increased in the ovariectomized mice after treatment with $17{\beta}$-estradiol. We report here for the first time that nesfatin-1/NUCB2 mRNA and protein express in the mouse uterus and its expression is regulated by estrogen secreted from the ovary, but not gonadotropin from the pituitary.

• Journal of Life Science
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• v.25 no.2
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• pp.223-230
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• 2015

Regulator of calcineurin 1 (RCAN1) is an endogenous calcineurin inhibitor that plays an important role in the pathogenesis of diseases related to the calcineurin-NFATc1 signaling pathway. The RCAN1-4 isoform is subject to NFATc1-dependent regulation. During receptor activator of nuclear factor kappa-B ligand (RANKL)-stimulated osteoclastogenesis, the calcineurin-NFATc1 pathway is critical. Because there is little information available on the role of RCAN1 in osteoclast differentiation, this study investigated whether changes in RCAN1 expression are related to the calcineurin-NFATc1 pathway and osteoclast differentiation. Mouse bone marrow monocytes (BMMs) were treated with 50 ng/ml of RANKL and M-CSF. Expression levels of NFATc1, calcineurin, and RCAN1 isoforms were determined using RT-PCR and Western blotting. Osteoclast differentiation was examined using tartrate-resistent acid phosphatase (TRAP) staining. To evaluate the effect of RCAN1 overexpression on osteoclastogenesis, cells were transfected with a mouse RCAN1-4 cDNA plasmid. After RANKL stimulation of BMMs, expression of NFATc1 and RCAN1 was increased at the mRNA and protein level, while calcineurin expression was unchanged. When the RCAN1-4 gene construct was transfected, the expression of RCAN1 protein was not increased despite several-fold increases in RCAN1-4 mRNA expression. Regardless of RANKL stimulation, over-expression of RCAN1-4 tended to reduce NFATc1 expression and knock-down of RCAN1 increase it. While BMMs transfected with the RCAN1-4 vector were differentiated into distinct osteoclasts, their phenotypes did not vary from those of mock controls. These results suggest that RCAN1 has a limited effect on the calcineurin-NFATc1 pathway during RANKL-stimulated osteoclast differentiation.

• Applied Microscopy
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• v.37 no.2
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• pp.93-102
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• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

• Journal of Nutrition and Health
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• v.42 no.5
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• pp.434-441
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• 2009

Ferroportin-1 (FPN) is a transporter protein that is known to mediate iron export from macrophages. The purpose of this study was to investigate the effect of copper on the regulation of FPN gene expression in J774 mouse macrophage cells. J774 cells were treated with various concentrations of $CuSO_4$ and RT-PCR analyses were performed to measure the steady-state levels of mRNAs for FPN and divalent metal transporter 1 (DMT1, an iron importer). Copper treatment significantly increased FPN mRNAs in a dose-dependent manner, but didn't change the levels of DMT1 mRNA. Experiments with transcriptional inhibitor actinomycin D (0.5 ${\mu}g$/mL) revealed that copper treatment did not affect the half-life of FPN mRNAs in J774 cells. On the other hand, results from luciferase reporter assays showed that copper directly stimulated the promoter activity of FPN. In summary, our data showed copper induced FPN mRNA of macrophages via a transcriptional rather than post-transcriptional mechanisms.

• Korean Journal of Veterinary Research
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• v.41 no.1
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• pp.1-6
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• 2001

Pituitary adenyl ate cyclase activating polypeptide (PACAP) was originally isolated from the ovine hypothalamus and stimulated cAMP production in anterior pituitary cells. It is known that PACAP stimulates cAMP accumulation and contributes to the spermatogenesis and steroidogenesis in rat testis. The principal aim of this study is to determinate the distribution of PACAP mRNA and protein in adult rat testis. For this study, we used in situ hybridization and immunohistochemistry techniques in adult rat testis. PACAP mRNA was stage specifically expressed in seminiferous tubules. Positive signals of PACAP mRNA were detected in the developing germ cells at stages HI-VII of the epithelial cycle. The strongest signals of PACAP mRNA and protein were detected in round spermatids at stages V to early VII of the cycle. These results demonstrate that PACAP which is synthesised in the developing germ cells contributes to the spermatogenesis in rat testis. Thus, we suggest that PACAP plays a critical role in the function of testis.

• The Korean Journal of Zoology
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• v.39 no.1
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• pp.47-53
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• 1996

We examined the expression of the heat shock proteins (HSPs) and glucose-regulated proteins (GRPs) during phorbol 1 2-myristate 1 3-acetate (PMA)-induced megakaryocytic differentiation of human er"'throleukemia K562 cells. PMA-treated K562 cells showed a cell growth arrest and alteration in morphology and patterns of gpllIa and c-myc expression, characteristic of megakaryocytic differentiation. During the megakaryocytic differentiation, HSP9OA, HSP9OB, and HSP28 mRNA and protein levels markedly decreased, while GRP78/B and GRP94 mRNA levels were enhanced. On the other hand, HSP7OA and HSP7OB mRNA levels were reduced, but HSP7O protein levels were not changed by PMA treatment. These results suggest specific roles for the HSPs and GRPs in K562 cell proliferation and megakaryocic differentiation.tion.

• 한국체육학회지인문사회과학편
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• v.55 no.4
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• pp.507-516
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• 2016

The purpose of this study is to identify the effects of PPARβ/δ over-expression on PGC-1α mRNA and protein stability after single bout of swimming exercise in mice skeletal muscle. Empty vector (EV) or PPARβ/δ was over-expressed in tibialis anterior(TA) using electroporation(EPO) technique to compare with non-treatment muscle(control; Con). TA muscles were dissected at 0h, 24h or 54h after termination of exercise. PGC-1α mRNA in Con, EV and PPARβ/δ over-expressed muscles were increased 6.8 fold (p<.001), 6.2 fold(p<.001) and 7.1 fold(p<.001), respectively, than sedentary(Sed) group at 0h after exercise and then reverted to Sed group levels at 24h and 54h after termination of exercise. PGC-1α and PGC-1α ubiquitination in EV treated muscles were increased 2.2 fold and 1.74 fold, respectively, than Sed group at 24h after termination of exercise, and then reverted to Sed group levels at 54h after termination of exercise. PGC-1α in PPARβ/δ over-expressed muscles at 24h and 54h after termination of exercise were increased 2.5 fold and 2.2 fold, respectively, than Sed group, but PGC-1α ubiquitination was not increased at 24h and 54h after termination of exercise. Our results indicate that PPARβ/δ over-expression does not increase PGC-1α mRNA stability, but increase PGC-1α protein stability through post-translation mechanism after termination of exercise.