• 한국응용과학기술학회지
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• 제40권6호
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• pp.1278-1288
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• 2023

헤스페리딘(Hesperidin, HD)은 다양한 식물체에 존재하는, 강한 항산화 기능을 가진 대표적인 flavonoid의 일종이다. 본 연구에서는 수용성 HD인 Hesperidin glucoside(HDG)가 가지는 세포손상 회복, 항염증 인자억제 및 melanin 생성억제 활성을 세포수준에서 비교하였다. HDG는 HD에 당전이 효소반응으로 제조되었으며, HD에 비해 20,000배 이상 수용해도가 증가되었다. HaCaT 세포주에 대한 세포독성은 HDG가 HD에 비해 월등히 낮았다. HD와 HDG는 모두 자외선 조사된 HaCaT 세포에서 세포생존율 회복효과를 나타내었다. 또한 HD와 HDG는 세포내 산화질소(NO), 종양괴사인자-α(TNF-α) 및 인터루킨-6(IL-6)과 같은 염증 매개체 및 cytokine을 감소시켰으며, HD 보다는 HDG의 효과가 다소 우수하였다. Melanoma B16F10 세포주를 이용한 melanin 형성능과 tyrosinase 저해활성을 측정한 결과, HD와 HDG 모두 효과를 나타내었으며 HDG가 약간 우수한 결과를 보였다. 결론적으로, HD의 당전이체인 HDG는 HD에 비해 동등이상의 세포손상 회복, 염증성 매개체 및 cytokine 억제능과 melanin 형성억제능을 나타내었으며, HDG의 높은 수용성과 낮은 세포독성 등의 특성은 다양한 분야에서의 용도를 확대시킬 수 있을 것으로 보인다.

• 동의생리병리학회지
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• 제26권6호
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• pp.908-914
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• 2012

To investigate the difference and change of ingredient and efficacy of Galgeun-tang (GGT) according to extraction solvent, water and 70% EtOH, the quantities of index components and the several in vitro activities of two kinds of GGT extract were compared. The contents of extracts were analyzed with HPLC. The biological activities such as anti-inflammatory and anti-obesity effects were measured through cell line-based in vitro assay. The cytotoxicity was measured by CCK-8 assay in RAW 264.7 cell, BEAS-2B cell, HaCaT cell and 3T3-L1 cell. We compared effects of two kinds of extract by measuring NO, PGE2, IL-6 and TNF-${\alpha}$ in RAW264.7 cell, RANTES in BEAS-2B cell, MDC and RANTES in HaCaT cell and GPDH activity and leptin level in differentiated 3T3-L1. 70% EtOH extract of GGT contained more compositions than water extract. The inhibitory effect of water extract of GGT on NO in RAW 264.7 cell and GPDH activity in 3T3-L1 was stronger than that of 70% EtOH. 70% EtOH has a stronger inhibitory effect on PGE2 in RAW264.7 cell, RANTES in BEAS-2B cell, MDC, RANTES in HaCaT cell, leptin in 3T3-L1 cell than water extraction. These results suggest that the ingredient and efficacy from 70% EtOH extract of GGT are more effective than water extract.

• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.13-24
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• 2001

Objective: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent $Ca^{2+}$-channel (1) P/Q-type $Ca^{2+}$-channel (2) N-type $Ca^{2+}$-channel (3) L-type $Ca^{2+}$-channel (4) T-type $Ca^{2+}$-channel (5) R-type $Ca^{2+}$-channel. The present study was done in order to investigate whether there is any difference in $Ca^{2+}$-channel distribution between activated and normally fertilized embryos. Methods: The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2+}$-channels in parthenogenetically activated 2-cell embryos by ethanol and $SrCl_2$ treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM $SrCl_2$ for 2h. Results: P/Q-type $Ca^{2+}$-channels and L-type $Ca^{2+}$-channels have been identified. Whereas, three type of $Ca^{2+}$-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. Conclusion: Activation by ethanol was faster than those by $SrCl_2$. However, there was difference in DAB staining of the embryos between ethanol and $SrCl_2$ treatment (87.7% and 54.1 %). Intensity of staining was also different between ethanol- and $SrCl_2$-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.

• The Korean Journal of Physiology and Pharmacology
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• 제12권4호
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• pp.177-183
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• 2008

The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.

• 대한본초학회지
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• 제34권1호
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• pp.99-107
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• 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.

• 생명과학회지
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• 제25권8호
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• pp.880-888
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• 2015

회향은 미나리과에 속하는 다년생식물인 Foeniculum vulgare Mill.의 성숙한 과실로서 항염, 진통, 피부노화방지 등의 효과를 가지고 있어 다양한 질환에 우수한 치료효과를 나타내는 것으로 알려져 왔다. 따라서 본 연구에서는 기능성이 우수한 회향 열매 추출물을 이용하여 피부 질환 치료제 및 피부장벽 기능 개선 소재로서의 적용가능성을 확인하였다. 이를 위해 각질형성세포주인 HaCaT에 회향 열매 메탄올 추출물과 그 분획물(hexane, methyl chloride, ethyl acetate, butanol)을 처리한 후, involucrin, loricrin, filaggrin의 발현과 hyaluronic acid의 생성 및 β-defensin -1, -2, -3, LL-37의 mRNA 발현을 측정하였다. 그 결과butanol 분획물 50 μg/ml을 24시간 동안 처리시 involucrin과 filaggrin단백질 발현을 각각122.8%, 105%로 유의하게 증가시킴을 확인하였다. Elisa assay를 통해 분석한 결과, ethyl acetate와 butanol 분획물은 대조군에 비해 hyaluronic acid의 생성을 각각17%, 11% 증가시켰으며, 이는 hyaluronic acid synthesis-1의 mRNA 발현 증가에 의한 것임을 확인하였다. 그러나 회향 열매 메탄올 추출물과 분획물 모두에서 β-defensin -1, -2, -3, LL-37의 mRNA 발현은 증가되지 않았다. 이러한 결과는 회향 열매 butanol 분획물이 각질형성세포에서 물리적 장벽을 형성하고 보습인자 조절을 통해 피부장벽 기능을 강화하는데 효과적임을 의미한다.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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• pp.36-36
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• 2002

In the present study, we investigated effects of extracellular zinc (Zn$\^$2＋/) on T-type Ca$\^$2＋/ channel isoforms (${\alpha}$lG, ${\alpha}$lH, and ${\alpha}$lI) stably expressed in HEK 293 cells. Ca$\^$2＋/ currents were measured using 10 mM Ca$\^$2＋/ as a charge carrier under whole cell-ruptured patch configuration. Zn$\^$2＋/ blocked the ${\alpha}$lH currents with a 100- and 200-fold higher potency (IC$\sub$50/ = 2.5 ${\mu}$M) when compared with those for blockade of the ${\alpha}$1G and ${\alpha}$1I currents, respectively.(omitted)

• Nutritional Sciences
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• 제8권4호
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• pp.242-249
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• 2005

It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

• 생명과학회지
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• 제28권3호
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• pp.345-350
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• 2018

이번 연구에서 우리는 6종의 미세조류 ; Phaeodacylum tricrnutum (PT), Chaetoceros gracilis (CG), Nanochloris oculata (NO), Pavlova lutheri (PL), Chlorella ellipsoidae (CE), and Scendedemus obliquus (SO)를 이용하여 탈모 예방 및 두피 개선효과를 살펴보았다. DPPH 라디칼 소거능 실험에서는 SO, CG, ST9 순으로 활성이 높은 것으로 나타났다. OH 소거능 실험 결과, CG (6.6~42.1%), ST9 (26.0~44.0%), SO (7.8~44.3%)으로 유의적인 효과를 보였다. 그외에도 세포증식실험에서는, SO가 인간표피세포주인 HaCaT 세포 증식을 촉진시켰으며, CG와 SO가 모유두세포주인 HFDPC 세포 증식을 촉진시키는 것으로 나타났다. 마지막으로 5-alpha reductase 저해능 실험결과, PT, CG, SO가 유의적으로 효소활성을 저해하였다. 따라서, 이번 연구에 사용한 6종의 미세조류들 중, 항산화활성, 세포증식 효과, 5-alpha reductase 저해능을 보인 몇가지 조류들이 탈모예방 및 두피개선을 위한 제품개발에 좋은 원료가 될 가능성이 있음을 제시한다.

• 한국치위생학회지
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• 제16권3호
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• pp.471-477
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• 2016

Objectives: Fluoride is widely used in the prevention and control of dental caries. The purpose of this study is to examine the biological effects of Sodium fluoride on the proliferation of oral normal cell in vitro(MDPC-23, HaCaT, HGF-1 cells). Methods: The proliferation of normal cells and the cyto-skeletal change of normal cells were assessed by WST-1 assay and F-actin stain assay. The statistical significances of the resulting data were analyzed using SPSS(Window 12.0). Results: The sodium fluoride(0-12 mM) treatment decreased the cell viability in a dose and time dependent manner: HaCaT(6 h): $100{\pm}0$, $98{\pm}0.39$, $82{\pm}2.68$, $75{\pm}0.83$, $69{\pm}1$, $67{\pm}1.42%$(p<0.005); HaCaT(24 h): $100{\pm}0$, $98{\pm}1.85$, $54{\pm}0.64$, $43{\pm}0.4$, $38{\pm}0.32$, $36{\pm}0.13%$(p<0.006), MDPC-23(6 h): $100{\pm}0$, $93{\pm}1.48$, $85{\pm}0.28$, $82{\pm}1.58$, $79{\pm}1.48$, $76{\pm}1.93%$(p<0.009); MDPC-23(24 h): $100{\pm}0$, $91{\pm}1.26$, $58{\pm}0.65$, $49{\pm}1$, $44{\pm}0.74$, $2{\pm}0.05%$(p<0.005), HGF-1(6 h): $100{\pm}0$, $97{\pm}2.93$, $89{\pm}5$, $71{\pm}5.42$, $58{\pm}4.82$, $43{\pm}3.47%$(p<0.009); HGF-1(24 h): $100{\pm}0$, $97{\pm}2.05$, $73{\pm}1.73$, $22{\pm}1.61$, $14{\pm}1.73$, $7{\pm}0.85%$(p<0.005). Thus, changes in cell morphology and disruption of filamentous(F)-actin organization were observed in higher concentration. Conclusions: These results suggest that higher concentrations of fluoride lead to a reduce the number of cells and morphology change of normal cell.