참당귀, 중국당귀, 일당귀 열수 추출물의 RAW 264.7 대식세포에서 IL-1β, TNF-α, iNOS 유전자 차등 발현 연구

한효상,홍성균,Han, Hyo-Sang,Hong, Seong-Gyun 한국디지털정책학회 2017 디지털융복합연구 Vol.15 No.11

We tried to analyze the inflammation reactions by treatment of AG, AS and AA in murine RAW 264.7 cells. To investigate the effect of AG, AS and AA on cell viability of RAW 264.7 cells, AG, AS and AA were treated for 24 h and MTS assay was performed. Cell viabilities were increased in $1,600{\mu}g/ml$ concentration by AS, AA and AG treatments, respectively. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by AG and AA treatment at a concentration of $200{\mu}g/ml$ in RAW 264.7 cells without Lipopolysaccharide (LPS) treatment. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by AG and AA 6 h treatment at a concentration of $200{\mu}g/ml$ with LPS treatment. In this study, we observed that AG, AS and AA show various activities on inflammation reaction depend on their treatment time. In the future, studies should be conducted to investigate the effects of AG, AS and AA on the various inflammatory responses of macrophages. Murine macrophage cell line인 RAW 264.7 대식세포에 참당귀(AG), 중국당귀(AS), 일당귀(AA)를 6시간과 24시간을 반응시킨 뒤 시간별로 나타나는 inflammation 반응을 확인하고자 하였다. LPS-induced RAW 264.7 대식세포에 AG, AS, AA를 6시간 반응 시킨 경우 $IL-1{\beta}$, $TNF-{\alpha}$, iNOS의 mRNA 발현량을 증가시켰고, AA에서 더 높은 Anti-inflammatory effect를 확인하였다. AG, AS, AA가 RAW 264.7 대식세포의 cell viability에 미치는 영향을 확인하기 위해 MTS Assay(24시간)를 수행한 결과 $1,600{\mu}g/ml$ 농도에서 모두 증가시켰다. AG, AS, AA가 LPS를 처리하지 않은 RAW 264.7 대식세포에 6시간동안 면역반응에 미치는 영향을 확인한 결과 AG와 AA를 $200{\mu}g/ml$ 농도로 처리한 RAW 264.7 대식세포에서 $IL-1{\beta}$, $TNF-{\alpha}$, iNOS의 발현이 증가되었다. 이 연구를 통해 AG, AS, AA는 RAW 264.7 대식세포에 LPS를 처리하였을 때 inflammation 반응을 촉진하며, 24시간 뒤 inflammation의 억제를 유도한 결과를 얻을 수 있었다. 향후 참당귀, 중국당귀, 일당귀 열수 추출물이 대식세포의 다양한 염증 반응 메커니즘에 미치는 작용을 확인하는 연구가 필요하다.

Lipopolysaccharide로 유발된 마우스 대식세포의 염증매개성 Cytokine 생성증가에 대한 참당귀 물추출물의 효능 연구

한효상 ( Hyo Sang Han ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.5

Objectives: The purpose of this study was to investigate the effects of Angelicae Gigantis Radix Water Extract(AG) on the production of proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide(LPS). Method: RAW 264.7 cells were cotreated with AG(50 and 100 ug/mL) and lipopolysaccharide(LPS; 1 ug/mL) for 24 hours. After 24 hour treatment, using Bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, IL-1β, IL-10, tumor necrosis factor-alpha(TNF-α), granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), interferon inducible protein-10(IP-10), leukemia inhibitory factor(LIF), lipopolysaccharide-induced chemokine(LIX), monocyte chemoattractant protein-1(MCP-1), macrophage colony-stimulating factor(M-CSF), macrophage inflammatory protein(MIP)-1α, MIP-1β, MIP-2, Regulated on Activation, Normal T cell Expressed and Secreted(RANTES) and vascular endothelial growth factor(VEGF) were measured. Result : AG significantly inhibited LPS-induced production of TNF-α, MIP-1α, G-CSF, RANTES, IL-10, and M-CSF from LPS-stimulated RAW 264.7 cells at the concentrations of 50 and 100 ug/mL. AG significantly inhibited LPS-induced production of MIP-1β, MIP-2, GM-CSF, and IL-6 from LPS-stimulated RAW 264.7 cells at the concentrations of 50 ug/mL. AG significantly inhibited LPS-induced production of VEGF from LPS-stimulated RAW 264.7 cells at the concentrations of 100 ug/mL. But AG did not show any significant effect on the production of MCP-1, LIF, LIX, IP-10 and IL-1β from LPS-induced RAW 264.7 cells. Conclusion : These results suggest that AG has anti-inflammatory effect related with its inhibition of proinflammatory mediators such as TNF-α, MIP-1α, G-CSF, RANTES, IL-10, MIP-1β, MIP-2, GM-CSF, IL-6, VEGF and M-CSF in LPS-induced macrophages.

현삼(玄參) 물추출물이 LPS로 유발된 대식세포의 염증인자에 미치는 영향

한효상 ( Hyo-sang Han ) 대한본초학회 2017 大韓本草學會誌 Vol.32 No.3

Objective : The purpose of this study was to investigate the effects of Scrophulariae Radix Water Extract (SR) on the production of inflammatory mediators in RAW 264.7 mouse macrophages cells induced by lipopolysaccharide (LPS). Method : We examined effect of Scrophulariae Radix Extract on the cell viability of mouse macrophages cells. Futhermore, After 24 hours treatment we investigated anti-inflammatory effect of Scrophulariae Radix Extract by the production of Bio-Plex cytokine assay, concentrations of various cytokines such NO, interleukin(IL)-1α, IL-3 and interferon inducible protein-10(IP-10). Result : No significant changes have been found in the mouse macrophge cell viability by the Scrophulariae Radix Extract at the concentration of 25, 50, 100 and 200 ㎍/㎖. The water extract of Scrophulariae Radix significantly inhibited the production of NO in the LPS-induced macrophage at the concentration of 25, 50, 100 and 200 ㎍/㎖. The water extract of Scrophulariae Radix significantly inhibited the production of IL-1α, IL-3 and IP-10 in the LPS-induced macrophage at the concentration of 50, 100 and 200 ㎍/㎖. Conclusion : The water extract of Scrophulariae Radix significantly inhibited the production of NO, IL-1α, IL-3 and IP-10 at the concentration of 50 ㎍/㎖ or higher in the LPS-induced macrophages with no changes in the cell viability of them. These results suggest that water extract of Scrophulariae Radix has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as IL-1α, IL-3 and IP-10 in the LPS-induced macrophages.

발효 애엽(艾葉) 추출물이 인간 간암세포주 HepG2 활성에 미치는 영향

한효상 ( Hyo Sang Han ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.3

Objective : The purpose of this study was to investigate the effect of fermented Artemisiae Argyi Folium(AAF) on some activities of human hepatoma cell, HepG2. Method : To investigate the effect of fermented Artemisiae Argyi Folium(AAF) activity on the human hepatoma cells, AAF extracts was fermented by Lactobacillus pentosus K34(AFL) and Sacchromyces cerevisiae STV89(AFS). And the effects of AFL or AFS on the activities of HepG2 cell, such as cell viability, nitric oxide(NO) production and reactive oxygen species(ROS) production, were tested. Result : Human Hepatoma Cells were incubated each for 3 hours and 24 hours. Human Hepatoma Cells treated with the extract was measured with MTT assay. Then AFL was found to be non-toxic at concentrations of 10 ug/mL(3h), 100 ug/mL(24h) or more. AFS was the same result at concentrations of more than 10 ug/mL. The extract increased ROS generation in Human Hepatoma Cells. AFL increased at concentrations of 100 ug/mL more (3h, also 10 ug/mL more) and 50 ug/mL(24h) and AFS increased both 50 ug/mL. In point of NO generation, AFL inhibited at concentrations of 10 ug/mL(3h) and 100 ug/mL(24h) more (3h, also 10 ug/mL more) and AFS also inhibited 50 ug/mL or more. Conclusion : AFL and AFS, obtained from Artemisiae Argyi Folium extracts by fermentation, reduced the NO production and increased ROS production in HepG2 cell, without cytotoxicity on HepG2 cell. The results suggested that AFL and AFS increased the immunological effects of Artemisiae Argyi Folium extracts.

꽃송이버섯 열수추출물이 HaCaT의 세포 연접 관련 유전자의 발현에 대한 영향

한효상 ( Hyo-sang Han ) 대한통합의학회 2021 대한통합의학회지 Vol.9 No.2

Purpose : Keratinocytes are the main cellular components involved in wound healing during re-epithelization and inflammation. Dysfunction of tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. The purpose of this study was to identify the various effects of a Sparassis crispa water extract (SC) on HaCaT cells and to investigate whether these effects might be applicable to human skin. Methods : We investigated the effectiveness of SC on cell HaCaT viability using MTS. The antioxidant effect of SC was analyzed by comparing the effectiveness of ABTS to that of the well-known antioxidant resveratrol. Reverse-transcription quantitative polymerase chain reaction (qRT-PCR) is the most widely applied method Quantitative RT-PCR analysis has shown that SC in HaCaT cells affects mRNA expression of tight-junction genes associated with skin moisturization. In addition, Wound healing is one of the most complex processes in the human body. It involves the spatial and temporal synchronization of a variety of cell types with distinct roles in the phases of hemostasis, inflammation, growth, re-epithelialization, and remodeling. wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells. Results : MTS analysis in HaCaT cells was found to be more cytotoxic in SC at a concentration of 0.5 ㎎/㎖. Compared to 100 μM resveratrol, 4 ㎎/㎖ SC exhibited similar or superior antioxidant effects. SC treatment in HaCaT cells reduced levels of claudin 1, claudin 3, claudin 4, claudin 6, claudin 7, claudin 8, ZO-1, ZO-2, JAM-A, occludin, and Tricellulin mRNA expression by about 1.13 times. Wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells and HaCaT cell migration was also reduced to 73.2 % by SC treatment. Conclusion : SC, which acts as an antioxidant, reduces oxidative stress and prevents aging of the skin. Further research is needed to address the effects of SC on human skin given the observed alteration of mRNA expression of tight-junction genes and the decreased the cell migration of HaCaT cells.

꽃송이버섯 추출물이 RAW 264.7 세포에서 TNF-α, iNOS, IL-1β 유전자 발현에 미치는 영향

한효상 ( Hyo-sang Han ) 대한통합의학회 2021 대한통합의학회지 Vol.9 No.1

Purpose : The purpose of this study was to examine the anti-inflammatory effects of Sparassis crispa (SC). SC is a well-known traditional herbal remedy and its mushroom is used for treatment of inflammation. Many diseases that are increasing recently have characteristics of inflammatory diseases. Researchers are finding bioactive substances from natural products that can promote treatment and prevention of inflammation. We investigated the effect of water extracted from SC on the expression of effector genes involved in the function of RAW 264.7 cells. Methods : Effects of RAW 264.7 cells on cell viability, antioxidation, and mRNA expression were examined using water extracts from SC. A 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of water extracts from SC on cell viability in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysaccharide (LPS) treatment and expression levels of inflammatory cytokine TNF-α, iNOS and IL-1β gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Results : The MTS assay was performed on RAW 264.7 cells after treatment with various concentrations of water extracts of SC. Treatment of RAW 264.7 cells with water extracts from SC and LPS at a concentration of 0.125, 0.5 ㎎/㎖ for twenty four hours promoted mRNA expression of TNF-α, iNOS and IL-1β. Conclusion : MTS assay was applied to RAW 264.7 cells after various concentrations of water extracts of SC. Through experimental demonstration of anti-oxidant and anti-inflammatory effects of water extracts from SC, we suggest that SC is a valuable material for the prevention and treatment of various inflammatory diseases.

애엽(艾葉) 발효 추출물의 면역활성에 관한 연구

한효상 ( Hyo Sang Han ),박완수 ( Wan Su Park ),이영종 ( Young Jong Lee ) 대한본초학회 2008 大韓本草學會誌 Vol.23 No.3

마우스 대식세포를 이용한 일당귀 물추출물의 항염효능 연구

한효상 ( Hyo Sang Han ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.6

Objectives: The purpose of this study was to investigate the effects of Angelicae acutilobae Radix Water Extract (AA) on the production of cytokines in RAW 264.7 cell stimulated with lipopolysaccharide (LPS). Method : RAW 264.7 cells were cotreated with AA (50 and 100 ㎍/mL) and lipopolysaccharide (LPS; 1 ㎍/mL) for 24 hours. After 24 hours treatment, using bead-based multiplex cytokine assay, concentrations of various cytokines such as interleukin(IL)-6, tumor necrosis factor-alpha(TNF-α) granulocyte colony-stimulating factor(G-CSF), granulocyte macrophage colony-stimulating factor(GM-CSF), and macrophage inflammatory protein(MIP)-1α were measured. Result : AA significantly inhibited LPS-induced production of IL-6 and MIP-1α from LPS-stimulated RAW 264.7 cells at the concentration of 50 ㎍/mL. AA significantly inhibited LPS-induced production of TNF-α from LPS-stimulated RAW 264.7 cells at the concentration of 100 ㎍/mL. AA significantly inhibited LPS-induced production of G-CSF and GM-CSF in RAW 264.7 cells at the concentrations of 50 and 100 ㎍/mL. Conclusion : These results suggest that AA has anti-inflammatory effect related with its inhibition of proinflammatory cytokines such as IL-6, TNF-α, G-CSF, GM-CSF, and MIP-1α in LPS-induced macrophages.

임신 말기 유도 분만에서 미소프로스톨 경질 투여와 옥시토신 정맥 투여의 효과와 안정성 비교

한효상 ( Hyo Sang Han ),이정재 ( Jeong Jae Lee ),안준모 ( Ahn Joon Mo ),정집광 ( Jib Kwang Chung ),이임순 ( Im Soon Lee ),이권해 ( Kwon Hae Lee ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.1

목적 : 임신말기의 유도분만에서 prostaglandin E1의 유도체인 미소프로스톨 경질 투여와 옥시토신 정맥 투여의 유도분만 효과 및 안정성을 비교하고자 본 연구를 시행하였다. 연구 방법 : 2000년 12월 1일부터 2002년 3월 31일까지 순천향 대학병원 분만실에 입원한 37주 이상의 임신부 중 유도분만의 금기증이 없는 63명의 임신부에게 25 킽의 미소프로스톨을 42명에게 3시간 간격으로 경질 투여하였고, 21명에게는 정맥으로 옥시토신을 분당 Objective : To compare the safety and efficacy of intravaginally administered misoprostol (PGE1 analogue) versus intravenously administered oxytocin for labor induction at term. Methods : Among 63 patients over 37 weeks, randomly selected 42 patients were

차가버섯 물 추출물의 추출온도에 따른 효능 비교 연구(II) -항산화 효능, 소염 및 항암 효과 연구-

박규천,한효상,이영종,Park, Kyu-Cheon,Han, Hyo-Sang,Lee, Young-Jong 대한본초학회 2007 大韓本草學會誌 Vol.22 No.4

Objectives : The present study purposed to compare the antioxidant effect, anti inflammatory effect and cancer cell multiplication inhibition effect of Fructificatio Inonoti Obliqui aqueous extract according to extraction temperature. Methods : We medicated animal models, which had experimental oxidation, with Fructificatio Inonoti Obliqui total extract and $50^{\circ}C$ low temperature leachate, and performed hematological analysis and blood chemical analysis with measuring SOD, GSH, catalase, NO and MDA content in the liver. In addition, we made comparative observation of anti inflammatory effect and anti-cancer effect. Results : Compared to the control group, both the group medicated with Fructificatio Inonoti Obliqui total extract and with $50^{\circ}C$ low-temperature leachate were found to decrease the number of thrombocytes in blood plasma and NO content while to increase SOD activity and catalase activity significantly. Both groups also showed anti-inflammatory effect against THP-1 cells and a multiplication inhibition effect against liver cancer cells and stomach cancer cells significantly. Conclusions : Both Fructificatio Inonoti Obliqui total extract and Fructificatio Inonoti Obliqui $50^{\circ}C$ low-temperature leachate have significant antioxidant effect, anti inflammatory effect and anti cancer effect.