• IMMUNE NETWORK
• /
• v.11 no.6
• /
• pp.399-405
• /
• 2011

Background: Endogenous uveitis is a chronic inflammatory eye disease of human, which frequently leads to blindness. Experimental autoimmune uveoretinitis (EAU) is an animal disease model of human endogenous uveitis and can be induced in susceptible animals by immunization with retinal antigens. EAU resembles the key immunological characteristics of human disease in that both are $CD4^+$ T-cell mediated diseases. Dendritic cells (DCs) are specialized antigen-presenting cells that are uniquely capable of activating naive T cells. Regulation of immune responses through modulation of DCs has thus been tried extensively. Recently our group reported that donor strain-derived immature DC pretreatment successfully controlled the adverse immune response during allogeneic transplantation. Methods: EAU was induced by immunization with human interphotoreceptor retinoid-binding protein (IRBP) $peptide_{1-20}$. Dendritic cells were differentiated from bone marrow in the presence of recombinant GM-CSF. Results: In this study, we used paraformaldehyde-fixed bone marrow-derived DCs to maintain them in an immature state. Pretreatment with fixed immature DCs, but not fixed mature DCs, ameliorated the disease progression of EAU by inhibiting uveitogenic $CD4^+$ T cell activation and differentiation. Conclusion: Application of iBMDC prepared according to the protocol of this study would provide an important treatment modality for the autoimmune diseases and transplantation rejection.

• Journal of Life Science
• /
• v.29 no.10
• /
• pp.1071-1079
• /
• 2019

In this study, we investigated the antioxidant and anti-aging activities of Schisandra chinensis seed fractions by adding them to UVB-irradiated HaCaT cells and analyzing the suppression of matrix metalloproteinases (MMPs). An electron-donating assay, ABTS radical scavenging assay, and hydrogen peroxide scavenging assay showed that the ethyl acetate fraction of S. chinensis seed (SCEAf) has scavenging activities. A collagenase inhibition activity assay showed that SCEAf had inhibitory effects of over 92.3% at $500{\mu}g/ml$ concentration. An MTT assay was performed to determine the cytotoxicity of SCEAf in HaCaT cells and the results showed that SCEAf increased cellular viability in a concentration-dependent manner. In addition, SCEAf was found to increase the viability of cells irradiatged by UVB $50mJ/cm^2$. To examine the anti-aging effects of SCEAf on HaCaT cells irradiated with UVB $50mJ/cm^2$, the expression of MMP-1 and -3 was analyzed by Western blotting and reverse-transcription polymerase chain reaction. The results showed that MMP-1 and -3 proteins and mRNA levels were downregulated in a concentration-dependent manner in response to SCEAf. These results suggest that SCEAf can prevent aging and alleviate aging symptoms by inhibiting collagenase activity in skin keratinocytes damaged by UVB. Therefore, S. chinensis seeds may have the potential for use as functional ingredients with anti-aging effects in the cosmetic and food industries.

• Restorative Dentistry and Endodontics
• /
• v.22 no.1
• /
• pp.374-387
• /
• 1997

The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.

• The Korea Journal of Herbology
• /
• v.34 no.1
• /
• pp.99-107
• /
• 2019

Objectives : Cudrania tricuspidata (C. tricuspidata) is well-known traditional herbal remedy and its root, leaf and fruit were used for treatment of inflammation, tumor and painkilling. However, effect of C. tricuspidata fruit on tight junction is still unknown. The aim of this research was to determine the effect of C. tricuspidata fruit extract on human keratinocyte HaCaT cells. Methods : The antioxidant effects of water extract of C. tricuspidata (WECT) and ethanol extract of C. tricuspidata (EECT) were analyzed by using an ABTS assay. To confirm the cytotoxicity of WECT and EECT, MTS assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Furthermore, dispase assay was used to investigate the alteration of cell-cell adhesion strength of EECT treated HaCaT cells. Results : WECT and EECT showed strong antioxidant activity. No obvious cytotoxicity was observed in both WECT and EECT until $2.0mg/m{\ell}$ concentration. The mRNA expression level of Claudin 6 were significantly increased by EECT treatment, whereas the WECT did not affect the expression of Claudin 6. Furthermore, EECT treatment enhances cell-cell adhesion strength. Conclusions : In this study, we investigated the physiological activities of the extracts of Cudrania tricuspidata fruit extracts on human keratinocytes by two different extraction methods. EECT might have an anti-aging activity on the skin by reducing oxidative stress. Moreover, it may be a useful ingredient in atopic dermatitis and skin-moisturizing, given its effects of altering Claudin 6 gene expression and enhancing cell-cell adhesion strength.

• Microbiology and Biotechnology Letters
• /
• v.41 no.3
• /
• pp.379-382
• /
• 2013

In order to investigate the anticancer activity of an anti-yeast substance (AYS), a proteoglycan produced by Rhanella aquatilis AY2000, the cytotoxicity of the AYS against cancer cells was determined in vitro. The AYS was not cytotoxic to the human Jurkat T cell or the mouse sarcoma 180 cell, but was cytotoxic to the human colon cancer TH20 cell. The AYS was increasingly cytotoxic against human colon cancer cells in a dose-dependent manner at range from 62.5 to 500 ${\mu}g/ml$. Anticancer activity by combination of the AYS and an anticancer agent was also determined. The anticancer agent combined with the AYS was shown to possess greater synergistic anticancer activity against human colon cancer cells, as compared with the anticancer agent alone.

• IMMUNE NETWORK
• /
• v.23 no.3
• /
• pp.29.1-29.23
• /
• 2023

Cholesterol (CL) is required for various biomolecular production processes, including those of cell membrane components. Therefore, to meet these needs, CL is converted into various derivatives. Among these derivatives is cholesterol sulfate (CS), a naturally produced CL derivative by the sulfotransferase family 2B1 (SULT2B1), which is widely present in human plasma. CS is involved in cell membrane stabilization, blood clotting, keratinocyte differentiation, and TCR nanocluster deformation. This study shows that treatment of T cells with CS resulted in the decreased surface expression of some surface T-cell proteins and reduced IL-2 release. Furthermore, T cells treated with CS significantly reduced lipid raft contents and membrane CLs. Surprisingly, using the electron microscope, we also observed that CS led to the disruption of T-cell microvilli, releasing small microvilli particles containing TCRs and other microvillar proteins. However, in vivo, T cells with CS showed aberrant migration to high endothelial venules and limited infiltrating splenic T-cell zones compared with the untreated T cells. Additionally, we observed significant alleviation of atopic dermatitis in mice injected with CS in the animal model. Based on these results, we conclude that CS is an immunosuppressive natural lipid that impairs TCR signaling by disrupting microvillar function in T cells, suggesting its usefulness as a therapeutic agent for alleviating T-cell-mediated hypersensitivity and a potential target for treating autoimmune diseases.

• The Korea Journal of Herbology
• /
• v.34 no.2
• /
• pp.25-32
• /
• 2019

Objectives : Gardeniae Fructus extract is used as a component of various cosmetics. However, the effect of the extract on the motility of keratinocytes has not been studied. The aim of this study is to investigate the inhibitory effect of ethanol extract of Gardeniae Fructus (GFET) or ethyl acetate extract of Gardeniae Fructus (GFEA) on oxidation and motility of human keratinocyte HaCaT cells. Methods : Antioxidant activity of Gardeniae Fructus extracts were determined by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. To investigate the cytotoxicity of Gardeniae Fructus extracts, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Cell migration assay was employed to determine the activity of Gardeniae Fructus extracts on motility of human keratinocyte HaCaT cells. Results : GFET and GFEA showed strong antioxidant activity. GFEA showed stronger cytotoxicity in HaCaT cells than GFET until $2.0mg/m{\ell}$ concentration. Cell migration assay demonstrated that GFET and GFEA decreased the motility of HaCaT cells. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by GFET or GFEA treatment. Conclusions : We investigated the physiological activities of the extracts of Gardeniae Fructus extracts on human keratinocytes by two different extraction methods. In addition, the mRNA expression level of claudin 8 among tight junction genes was significantly reduced by either GFET or GFEA treatment. This study provides basic information on the application of Gardeniae Fructus extract to cosmetics component.

• Korean Journal of Medicinal Crop Science
• /
• v.12 no.1
• /
• pp.36-42
• /
• 2004

The immune activation and anticancer activities of the water and ethanol extracts from Artemisia capillaris Thunb. were studied. The growth of human hepatocarcinoma and human gastric cancer cell was inhibited by the addition of $1.0\;mg/m{\ell}$ of the water extract, by about 77% and 95%, respectively. The growth of human breast cancer cells was also inhibited by addition of $0.5\;mg/m{\ell}$ of both water and ethanol extracts by 88%. The growth of human normal lung cell, HEL299 was inhibited by 15% indicating very low cytotoxicity of both extracts. Overall selectivity of the both extracts on several human cancer cell line was over 2.5. The growth of both human B and T cells was enhanced up to 1.6 to 2.1 times by adding the ethanol extracts. The secretion of cytokines, $TNF-{\alpha}$ and IL-6, from human B cells was also increased showing $68\;pg/m{\ell}$ and $67\;pg/m{\ell}$, respectively, compared to $35{\sim}40\;pg/m{\ell}$ of the control. In terms of the immune activity, there was not much difference between water and ethanol extracts of Artemisia capillaris Thunb. It implies that the extraction solvent could not differ the biological activities of the extracts. Based on these results, Artemisia capillaris Thunb. can be developed into a potentially useful cancer chemoprentive agent.

• Biomolecules & Therapeutics
• /
• v.24 no.6
• /
• pp.616-622
• /
• 2016

Baicalein (5,6,7-trihydroxy-2-phenyl-chromen-4-one) is a flavone, a type of flavonoid, originally isolated from the roots of Scutellaria baicalensis. This study evaluated the protective effects of baicalein against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) radiation in a human keratinocyte cell line (HaCaT). Baicalein absorbed light within the wavelength range of UVB. In addition, baicalein decreased the level of intracellular reactive oxygen species (ROS) in response to UVB radiation. Baicalein protected cells against UVB radiation-induced DNA breaks, 8-isoprostane generation and protein modification in HaCaT cells. Furthermore, baicalein suppressed the apoptotic cell death by UVB radiation. These findings suggest that baicalein protected HaCaT cells against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging ROS.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.20 no.3
• /
• pp.14-28
• /
• 2007

Objective : Hwagi-Jokyeong-Tang (化氣調經湯, HJT) was described in DongeuiBogam(東醫寶鑑). This remedy has been used to treat patients with Naryeok, which is similar as tuberculous cervical lymphadenitis in western medicine. Methods : In this study, the present author investigated the effects of HJT on on Human HaCaT keratinocyte and malignant melanoma cells such as SK-MEL-2 and B16F10 in terms of cell viabilities, proliferations, DPPH free radical scavenging activities, oxygen free radical productions and inhibitory action on elastase activities. Results : HJT acceleated proliferation of HaCaT keratinocytes dose-dependantly. HJT also prevented cell death of HaCaT induced by Hydrogen peroxide, which products oxygen free radicals. On the contrary, HJT did not affect proliferations of SK-MEL-2 or B16F10. In addition, HJT was shown to have DPPH free radical scavenging activities and also have inhibitory effects on elastase activities too. On the fluorescent examinations, the present author know that HJT did not affect production levels of oxygen free radicals in malignant melanoma cell, SK-MEL-2. Conclusions : These results suggest that HJT has possibilities of usage for functional cosmetics which have skin regeneration or prevention from skin tissue injury.