• Toxicological Research
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• v.22 no.3
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• pp.283-291
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• 2006

The survival and growth of epithelial cells depends on adhesion to the extracellular matrix. An adhesion signal may regulate the initiation of differentiation, since epidermal keratinocytes differentiate as they leave the basement membrane. A metabolically dead cornified cell envelope is the end point of epidermal differentiation so that this process may be viewed as a specialized form of programmed cell death. In order to investigate the precise cellular signaling events loading to terminal differentiation of keratinocytes, we have utilized HaCaT cells to monitor the biological consequences of $Ca^{2+}$ stimulation and numerous downstream signaling pathways, including activation of the extracellular signal-regulated protein kinase(ERK) pathway and activation of phosphatidylinositol 3-kinase(PI3K). The results presented in this study show that $Ca^{2+}$ function as potent agents for the differentiation of HaCaT keratinocytes, and this differentiation depends or the activation of ERK, Protein kinase B(PKB) and p70 ribosomal protein S6 kinase(p70S6K). Finally, the results show that the expression of Activator protein 1(AP-1; c-Jun and c-Fos) increased following $Ca^{2+}$-mediated differentiation of HaCaT cells, suggesting that ERK-mediated AP-1 expression is critical for initiating the terminal differentiation of keratinocytes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.4
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• pp.805-817
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• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.

• Biomolecules & Therapeutics
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• v.24 no.6
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• pp.595-603
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• 2016

(E)-3-Phenyl-1-(2-pyrrolyl)-2-propenone (PPP) is a pyrrole derivative of chalcone, in which the B-ring of chalcone linked to ${\beta}$-carbon is replaced by pyrrole group. While pyrrole has been studied for possible Src inhibition activity, chalcone, especially the substituents on the B-ring, has shown pharmaceutical, anti-inflammatory, and anti-oxidant properties via inhibition of NF-${\kappa}B$ activity. Our study is aimed to investigate whether this novel synthetic compound retains or enhances the pharmaceutically beneficial activities from the both structures. For this purpose, inflammatory responses of lipopolysaccharide (LPS)-treated RAW264.7 cells were analyzed. Nitric oxide (NO) production, inducible NO synthase (iNOS) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) mRNA expression, and the intracellular inflammatory signaling cascade were measured. Interestingly, PPP strongly inhibited NO release in a dose-dependent manner. To further investigate this anti-inflammatory activity, we identified molecular pathways by immunoblot analyses of nuclear fractions and whole cell lysates prepared from LPS-stimulated RAW264.7 cells with or without PPP pretreatment. The nuclear levels of p50, c-Jun, and c-Fos were significantly inhibited when cells were exposed to PPP. Moreover, according to the luciferase reporter gene assay after cotransfection with either TRIF or MyD88 in HEK293 cells, NF-${\kappa}B$-mediated luciferase activity dose-dependently diminished. Additionally, it was confirmed that PPP dampens the upstream signaling cascade of NF-${\kappa}B$ and AP-1 activation. Thus, PPP inhibited Syk, Src, and TAK1 activities induced by LPS or induced by overexpression of these genes. Therefore, our results suggest that PPP displays anti-inflammatory activity via inhibition of Syk, Src, and TAK1 activity, which may be developed as a novel anti-inflammatory drug.

• Journal of Life Science
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• v.28 no.1
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• pp.68-77
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• 2018

The purpose of this study was to isolate the probiotic lactic acid bacteria, and verify the possibility of the final selection strain as probiotic material. For screening of biogenic amines non-producing microorganisms, 42 lactic acid bacteria were isolated from various berries, extract and vinegar grown in Sunchang. Isolates were investigated for various physiological activities such as extracellular enzyme, antimicrobial and antioxidant activities, and 5 isolates were firstly screened. SBB07 was finally selected by analyzing the biogenic amine, and named Lactobacillus brevis SBB07 by 16S rRNA sequencing analysis. Next, SBB07 was assayed for their survival ability when exposed to acidic and bile conditions as well as heat and antibiotic resistance. As a result, SBB07 showed more than 86% and 54% higher survival rate in acidic condition at pH 2.0 and bile resistance with 0.5% oxgall. In addition, SBB07 showed a survival rate of more than 113% in $60^{\circ}C$, and also confirmed that it has resistant to various antibiotics. As a result of confirming the possibility of prebiotics, SBB07 showed the best utilization of GOS as a prebiotic substrate, and utilization of FOS and inulin were also high. These results suggest that SBB07 have good potential for application as probiotic lactic acid bacteria.

• BMB Reports
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• v.53 no.4
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• pp.218-222
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• 2020

Excessive and hyperactive osteoclast activity causes bone diseases such as osteoporosis and periodontitis. Thus, the regulation of osteoclast differentiation has clinical implications. We recently reported that dehydrocostus lactone (DL) inhibits osteoclast differentiation by regulating a nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), but the underlying mechanism remains to be elucidated. Here we demonstrated that DL inhibits NFATc1 by regulating nuclear factor-κB (NF-κB), activator protein-1 (AP-1), and nuclear factor-erythroid 2-related factor 2 (Nrf2). DL attenuated IκBα phosphorylation and p65 nuclear translocation as well as decreased the expression of NF-κB target genes and c-Fos. It also inhibited c-Jun N-terminal kinase (JNK) but not p38 or extracellular signal-regulated kinase. The reporter assay revealed that DL inhibits NF-κB and AP-1 activation. In addition, DL reduced reactive oxygen species either by scavenging them or by activating Nrf2. The DL inhibition of NFATc1 expression and osteoclast differentiation was less effective in Nrf2-deficient cells. Collectively, these results suggest that DL regulates NFATc1 by inhibiting NF-κB and AP-1 via down-regulation of IκB kinase and JNK as well as by activating Nrf2, and thereby attenuates osteoclast differentiation.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.2
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• pp.23-39
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• 2014

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

• International Journal of Oral Biology
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• v.34 no.1
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• pp.1-6
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• 2009

The purpose of the present study was to examine the role of peripheral nitric oxide (NO) pathways in the onset of interleukin (IL)-1$\beta$-induced mechanical allodynia in the orofacial area. Experiments were carried out on male Sprague-Dawley rats weighing 230-280 gm and surgical procedures were performed under pentobarbital sodium (40 mg/kg, i.p.). Under anesthesia, a polyethylene tube (PE10) was implanted into the subcutaneous area of one vibrissa pad, which enabled the injection of IL-1$\beta$ or other chemicals. We subcutaneously injected 50 ${\mu}L$ of IL-1$\beta$ into a vibrissa pad through the implanted polyethylene tube with a 100 ${\mu}L$ Hamilton syringe. After the administration of 0.01, 0.1, 1, or 10 pg of IL-1$\beta$, withdrawal behavioral responses were examined. The subcutaneous injection of saline had no effects on the air-puff thresholds. Following the subcutaneous injection of 0.01, 0.1, 1, or 10 pg of IL-1$\beta$, the threshold of air puffs decreased significantly to 12 $\pm$ 3, 7 $\pm$ 2, 5 $\pm$ 1, or 5 $\pm$ 1 psi, respectively, in a dose dependent manner. Pretreatment with L-NAME, a nitric oxide synthase (NOS) inhibitor, blocked IL-1$\beta$-induced mechanical allodynia. However, neither D-NAME, an inactive isomer of L-NAME, nor vehicle affected the IL-1$\beta$-induced mechanical allodynia. Subcutaneous injection of IL-1$\beta$ increased the number of c-fos-like immunoreactive neurons, whereas pretreatment with L-NAME decreased this number, in the trigeminal caudal nucleus. These results suggest that pro-inflammatory cytokines and NO are important contributors to the pathogenesis of persistent and exaggerated IL-1$\beta$-induced pain states. Based on these observations, peripheral application of NOS inhibitors may be of therapeutic value in treating pain disorders in the clinic.

• Animal Bioscience
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• v.35 no.7
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• pp.964-974
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry and economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for studies on HPAIV resistance. Therefore, in this study, we investigated gene expression related to the mitogen-activated protein kinase (MAPK) signaling pathway by comparing non-infected, HPAI-infected resistant, and susceptible Ri chicken lines. Methods: Resistant (Mx/A; BF2/B21) and susceptible Ri chickens (Mx/G; BF2/B13) were selected by genotyping the Mx and BF2 genes. Then, the tracheal tissues of non-infected and HPAIV H5N1 infected chickens were collected for RNA sequencing. Results: A gene set overlapping test between the analyzed differentially expressed genes (DEGs) and functionally categorized genes was performed, including biological processes of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. A total of 1,794 DEGs were observed between control and H5N1-infected resistant Ri chickens, 432 DEGs between control and infected susceptible Ri chickens, and 1,202 DEGs between infected susceptible and infected resistant Ri chickens. The expression levels of MAPK signaling pathway-related genes (including MyD88, NF-κB, AP-1, c-fos, Jun, JunD, MAX, c-Myc), cytokines (IL-1β, IL-6, IL-8), type I interferons (IFN-α, IFN-β), and IFN-stimulated genes (Mx1, CCL19, OASL, and PRK) were higher in H5N1-infected than in non-infected resistant Ri chickens. MyD88, Jun, JunD, MAX, cytokines, chemokines, IFNs, and IFN-stimulated expressed genes were higher in resistant-infected than in susceptible-infected Ri chickens. Conclusion: Resistant Ri chickens showed higher antiviral activity compared to susceptible Ri chickens, and H5N1-infected resistant Ri chickens had immune responses and antiviral activity (cytokines, chemokines, interferons, and IFN-stimulated genes), which may have been induced through the MAPK signaling pathway in response to H5N1 infection.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

• Journal of Acupuncture Research
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• v.31 no.2
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• pp.119-134
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• 2014

Objectives : The purpose of this study was to investigate the analgesic effects of filiform acupuncture and laser acupuncture at the points of $TE_3{\cdot}GB_{41}$ in neuropathic pain induced rats. Methods : Neuropathic pain in rats was induced by partial resection of the tibial and sural nerves. Three weeks after the neuropathic surgery, each of the experimental groups(AT, LAT, AT + LAT and LAT + AT) was injected at the $TE_3{\cdot}GB_{41}$ twice per week for three weeks. Results : 1. All of the experimental groups(AT, LAT, AT + LAT and LAT + AT) showed a significant decrease in the plantar withdrawal response of allodynia and the thermal allodynia as compared with the control group. During the early phase, the AT and AT + LAT groups have been marked as more significant than the LAT + AT and LAT groups. 2. The expression of c-Fos significantly decreased in the LAT and LAT + AT groups as compared with the control group. 3. The LAT + AT group showed a significant decrease in Bax as compared with the control group. In each experimental groups(AT, LAT, AT + LAT and LAT + AT), Bcl-2 increased and Bax/Bcl-2 ratio decreased as compared with the control group. 4. The LAT, AT + LAT and LAT + AT groups showed a significant increase in mGluR5 as compared with the control group. Conclusions : These results represented that the filiform acupuncture and laser acupuncture at the $TE_3{\cdot}GB_{41}$ exerted anti-apoptotic and neuroprotective effects on the model of neuropathic pain, thereby suggesting that they should be available for decreasing mechanical allodynia.