• 한국산학기술학회논문지
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• 제12권10호
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• pp.4378-4384
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• 2011

본 연구는 백지 에탄올추출물의 항산화 활성을 알아보기 위하여 총 포리페놀 함량, 총 플라보노이드 함량, 전자공여능을 독립적으로 3회 이상 반복 실시하여 측정 하였고, brown guinea pig(450~500g)의 등 부위에 1500mJ/$cm^2$ 광량의 자외선 조사로 유발시킨 피부(6areas per group)에 1일 2회, 주 5일, 매회 $30{\mu}{\ell}$씩 총 5주간 시료를 도포한 후 염산 케타민으로 마취, 시료를 도포한 피부 부위를 직경12mm의 biopsy punch로 절취하여 10%의 중성 포르말린 용액에 12시간 실온에서 고정한 후 Hematoxylin and eosin(H&E) 염색으로 표피 및 진 조직을 관찰하였다. 백지 에탄올추출물의 총 폴리페놀 함량과 총 플라보노이드 함량을 측정한 결과, 각각 20.7mg/g, 19.5mg/g으로 확인되어 항산화물질 함량이 양호하게 나타났다. 전자공여능을 측정한 결과, 백지 에탄올추출물은 농도가 증가함에 따라 전자공여능 또한 유의하게 증가하였고, $500{\mu}g/m{\ell}$과 $1000{\mu}g/m{\ell}$농도에서 각각 14.8%, 19.8%의 전자공여능을 보여 양성대조군으로 사용한 Dibutylated hydroxytoluene(BHT)보다 공여능이 낮았으나 의미 있는 항산화 효과를 보였다. H&E 염색 결과, 대조군의 표피에서 경미한 비후도가 나타났으나, 백지 에탄올추출물 도포군은 피부조직 배열이 규칙적이며 염증관련 세포 침윤 등의 별다른 이상 징후는 발견되지 않았다. 이상의 결과를 종합하면 백지 에탄올추출물은 천연 항산화제로서 이용가치가 있을 것으로 판단된다.

• The Journal of Korean Physical Therapy
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• 제20권1호
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• pp.57-65
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• 2008

Purpose: Heat shock proteins (HSPs) are a group of proteins that are activated when cells are exposed to a variety of environmental stresses, such as infection, inflammation, exposure to toxins, starvation, hypoxia, brain injury, or water deprivation. The activation of HSPs by environmental stress plays a key role in signal transduction, including cytoprotection, molecular chaperone, anti-apoptotic effect, and anti-aging effects. However, the precise mechanism for the action of small HSPs, such as HSP27 and mitogen-activated protein kinases (MAPKs: extracellular-regulated protein kinase 1/2 (ERK1/2), p38MAPK, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), is not completely understood, particularly in application of cell stimulators including platelet-derived growth factor (PDGF), angiotensin II (AngII), tumor necrosis factor $\alpha$ (TNF$\alpha$), and $H_2O_2$. This study examined the relationship between stimulators-induced enzymatic activity of HSP27 and MAPKs from rat smooth and skeletal muscles. Methods: 2-dimensional electrophoresis (2DE) and matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-TOF/TOF) analysis were used to identify HSP27 from the intact vascular smooth and skeletal muscles. Three isoforms of HSP27 were detected on silver-stained gels of the whole protein extracts from the rat aortic smooth and skeletal muscle strips. Results: The expression of PDGF, AngII, TNF$\alpha$, and $H_2O_2$-induced activation of HSP27, p38MAPK, ERK1/2, and SAPK/JNK was higher in the smooth muscle cells than the control. SB203580 (30${\mu}$M), a p38MAPK inhibitor, increased the level of HSP27 phosphorylation induced by stimulators in smooth muscle cells. Furthermore, the age-related and starvation-induced activation of HSP27 was higher in skeletal muscle cells (L6 myoblast cell lines) and muscle strips than the control. Conclusion: These results suggest, in part, that the activity of HSP27 and MAPKs affect stressors, such as PDGF, AngII, TNF$\alpha$, $H_2O_2$, and starvation in rat smooth and skeletal muscles. However, more systemic research will be needed into physical therapy, including thermotherapy, electrotherapy, radiotherapy and others.

• Journal of Applied Biological Chemistry
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• 제59권4호
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• pp.365-372
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• 2016

본 연구에서는 150 mM HCl/60 % ethanol로 급성 위염을 유발한 마우스에서 증숙 시간에 따른 홍삼의 위염 보효 효과에 대해 살펴보고자 하였다. 백삼과 홍삼의 증숙 시간에 따른 성분을 분석한 결과 사포닌, total polyphenol과 total flavonoid의 총 함량이 증숙 시간에 따라 증가하였고 6시간 증숙한 홍삼에서 가장 높은 함량을 보였다. 또한 1,1-diphenyl-2-picrylhydrazyl와 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid radical 소거능 실험을 통해 항산화 활성을 측정한 결과 RG 6에서 가장 높은 활성을 나타냈다. In vitro 실험 결과를 바탕으로 시료를 선택하였고 in vivo 실험을 진행하였다. 급성 위염 마우스 모델에 백삼과 6시간 증숙한 홍삼을 투여하였을 때, RG 6에서 위 점막 손상의 개선을 육안적으로 확인할 수 있었으며, 혈액에서 측정한 ROS 수치도 대조군에 비해 유의적인 감소를 보였다. 또한 염증성 사이토카인을 확인한 결과 대조군에 비해 RG6에서 감소하는 경향을 보였다. 이러한 결과들을 종합해 볼 때 증숙 시간에 따른 홍삼은 급성 위염 유발 마우스 모델에서 위염 보호 효과가 있는 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권1호
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• pp.43-50
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• 2015

It has been shown that the extracts including eupatilin and quercetin-3-${\beta}$-D-glucuronopyranoside had mucoprotective effects on the esophagus and stomach through their antioxidant activities. This study was designed to investigate the anti-inflammatory effect of these flavonoid compounds in an animal model of inflammatory bowel disease induced by 2,4,6-trinitrobenzene sulfonic acid. Experimental colitis was induced by intracolonic administration of 2,4,6-trinitrobenzene sulfonic acid. Extracts including eupatilin or quercetin-3-${\beta}$-D-glucuronopyranoside were orally administered to animals 48, 24, and 1 h prior to the induction of colitis and then again 24 h later. The animals were sacrificed 48 h after by 2,4,6-trinitrobenzene sulfonic acid treatment and the macroscopic appearance of the colonic lesions was scored in a blinded manner on a scale of 1 to 10. The inflammatory response to colitis induction was assessed by measuring myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, total glutathione levels, and malondialdehyde concentrations in the colon. The results indicated that extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside dose-dependently improved the morphology of the lesions induced by 2,4,6-trinitrobenzene sulfonic acid and reduced the ulcer index accordingly. In addition, rats receiving extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside showed significantly decreased levels of mucosal myeloperoxidase activity, nitric oxide production, tumor necrosis factor-${\alpha}$ expression, and malondialdehyde levels, and increased total glutathione levels. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside ameliorated the inflammatory response and colonic injury in acute colitis by decreasing oxidative stress and neutrophil activation. Extracts including eupatilin and extracts including quercetin-3-${\beta}$-D-glucuronopyranoside may inhibit acute colitis.

• Nutrition Research and Practice
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• 제15권6호
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• pp.673-685
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• 2021

BACKGROUND/OBJECTIVES: Obesity is associated with the impaired regulation of T cells characterized by increased numbers of Th1 and Th17 cells and the dysregulation of vitamin D metabolism. Both obesity and vitamin D have been reported to affect autophagy; however, a limited number of studies have investigated the effects of vitamin D on T cell autophagy in obese mice. Therefore, we aimed to determine whether in vitro treatment with vitamin D affects the proliferation, function, and autophagy of T cells from obese and control mice. MATERIALS/METHODS: Five-week-old male C57BL/6 mice were fed control or high-fat diets (10% or 45% kcal fat: CON or HFDs, respectively) for 12 weeks. Purified T cells were stimulated with anti-CD3 and anti-CD28 monoclonal antibodies and cultured with either 10 nM 1,25(OH)₂D₃ or 0.1% ethanol (vehicle control). The proliferative response; expression of CD25, Foxp3, RORγt, and autophagy-related proteins (LC3A/B, SQSTM1/P62, BECLIN-1, ATG12); and the production of interferon (IFN)-γ, interleukin (IL)-4, IL-17A, and IL-10 by T cells were measured. RESULTS: Compared with the CON group, T cell proliferation tended to be lower, and the production of IFN-γ was higher in the HFD group. IL-17A production was reduced by 1,25(OH)2D₃ treatment in both groups. The LC3 II/I ratio was higher in the HFD group than the CON group, but P62 did not differ. We observed no effect of vitamin D treatment on T cell autophagy. CONCLUSIONS: Our findings suggest that diet-induced obesity may impair the function and inhibit autophagy of T cells, possibly leading to the dysregulation of T cell homeostasis, which may be behind the aggravation of inflammation commonly observed in obesity.

• 대한임상검사과학회지
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• 제51권2호
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• pp.214-220
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• 2019

황련해독탕은 한방산업에서 염증관련 질환에 다빈도로 사용되는 처방이다. 본 연구의 목적은 전통적 한약인 황련해독탕의 항균활성을 확인하고 지표성분을 선정하는 것이다. 황련해독탕의 항균활성을 측정하기 위하여 디스크 확산법과 최소억제농도법을 사용하였다. 사용균주는 Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 23724였으며 대조군으로 oxacillin과 ciprofloxacin을 이용하여 황련해독탕의 항균활성을 확인하였다. 결과적으로 디스크 확산법에서는 S.aureus ATCC 25923에서 황련해독탕 20 mg과 10 mg은 $11.7{\pm}1.3mm$, $8.7{\pm}0.7mm$의 억제환을 형성하였으며 E.coli ATCC 23724에서는 황련해독탕 20 mg이 $8.0{\pm}0.0mm$의 억제환을 형성하였다. 최소억제농도는 S.aureus ATCC 25923가 31.25 mg/mL, E.coli ATCC 23724 에서는 125.0 mg/mL였다. 표준화를 위한 HPLC 정량분석 결과, 황련해독탕은 berberine을 16.55 mg/g 함유하고 있었으며 geniposide를 81.85 mg/g 함유하고 있었다. 천연물을 이용하여 항생제를 개발할 경우에는 내성세균을 억제하고 부작용을 줄일 수 있다는 장점이 있다. 이러한 점을 착안하여 지속적으로 연구를 진행한다면 한방산업과 더불어 여러 산업에서의 발전을 기대할 수 있을 것이다.

• The Korean Journal of Physiology and Pharmacology
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• 제25권1호
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• pp.59-68
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• 2021

Arterial thrombosis and its associated diseases are considered to constitute a major healthcare problem. Arterial thrombosis, defined as blood clot formation in an artery that interrupts blood circulation, is associated with many cardiovascular diseases. Oxidative stress is one of many important factors that aggravates the pathophysiological process of arterial thrombosis. Apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ref-1) has a multifunctional role in cells that includes the regulation of oxidative stress and anti-inflammatory function. The aim of this study was to investigate the therapeutic effect of adenovirus-mediated Ref-1 overexpression on arterial thrombosis induced by 60% FeCl3 solution in rats. Blood flow was measured to detect the time to occlusion, thrombus formation was detected by hematoxylin and eosin staining, reactive oxygen species (ROS) levels were detected by high-performance liquid chromatography, and the expression of tissue factor and other proteins was detected by Western blot. FeCl3 aggravated thrombus formation in carotid arteries and reduced the time to artery occlusion. Ref-1 significantly delayed arterial obstruction via the inhibition of thrombus formation, especially by downregulating tissue factor expression through the Akt-GSK3β-NF-κB signaling pathway. Ref1 also reduced the expression of vascular inflammation markers ICAM-1 and VCAM-1, and reduced the level of ROS that contributed to thrombus formation. The results showed that adenovirus-mediated Ref-1 overexpression reduced thrombus formation in the rat carotid artery. In summary, Ref-1 overexpression had anti-thrombotic effects in a carotid artery thrombosis model and could be a target for the treatment of arterial thrombosis.

• 한방안이비인후피부과학회지
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• 제35권4호
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• pp.75-94
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• 2022

Objectives : To identify the active ingredient of Poncirus Trifoliata Immaturus and to explore the mechanism expected to potentially act on dermatitis accompanied by pruritus. Methods : We conducted the network pharmacological analysis. We selected effective ingredients among the active compounds of Poinciri Fructus Immaturus. We found the target protein of the selected active ingredient, disease(dermatitis accompanied by pruritus) and fexofenadine. Then we established the network between the proteins which Poinciri Fructus Immaturus and fexofenadine intersected with disease respectively, and the coregene was also extracted. After that, the active pathways in the human body involving the groups and coregenes were searched. Results : Total of 7 active ingredients were selected, and 202 target proteins were collected. There were 756 proteins related to inflammatory skin disease accompanied by pruritus, and 75 proteins were related to fexofenadine. 42 proteins crossed by Poinciri Fructus Immaturus with a disease, and 31 proteins crossed by fexofenadine with a disease. 12 proteins were found as a coregene from the proteins that cross Poinciri Fructus Immaturus and disease. Coregenes are involved in 'Nitric-oxide synthase regulator activity', 'Epidermal growth factor receptor signaling pathway'. 2 groups that extracted are invloved in 'Fc receptor signaling pathway', 'Central carbon metabolism in cancer', 'Phosphatidylinositol 3-kinase complex, class IB', and 'omega-hydroxylase P450 pathway'. Conclusion : It is expected that Poinciri Fructus Immaturus will be able to show direct or indirect anti-pruritus and anti-inflammatory effects on skin inflammation accompanied by pruritus in the future. And it is also expected to have a synergy effect with fexofenadine on skin disease.

• IMMUNE NETWORK
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• 제20권4호
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• pp.32.1-32.15
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• 2020

Influenza virus is the major cause of seasonal and pandemic flu. Currently, oseltamivir, a potent and selective inhibitor of neuraminidase of influenza A and B viruses, is the drug of choice for treating patients with influenza virus infection. However, recent emergence of oseltamivir-resistant influenza viruses has limited its efficacy. Morin hydrate (3,5,7,2',4'-pentahydroxyflavone) is a flavonoid isolated from Morus alba L. It has antioxidant, anti-inflammatory, neuroprotective, and anticancer effects partly by the inhibition of the NF-κB signaling pathway. However, its effects on influenza virus have not been studied. We evaluated the antiviral activity of morin hydrate against influenza A/Puerto Rico/8/1934 (A/PR/8; H1N1) and oseltamivir-resistant A/PR/8 influenza viruses in vitro. To determine its mode of action, we carried out time course experiments, and time of addition, hemolysis inhibition, and hemagglutination assays. The effects of the co-administration of morin hydrate and oseltamivir were assessed using the murine model of A/PR/8 infection. We found that morin hydrate reduced hemagglutination by A/PR/8 in vitro. It alleviated the symptoms of A/PR/8-infection, and reduced the levels of pro-inflammatory cytokines and chemokines, such as TNF-α and CCL2, in infected mice. Co-administration of morin hydrate and oseltamivir phosphate reduced the virus titers and attenuated pulmonary inflammation. Our results suggest that morin hydrate exhibits antiviral activity by inhibiting the entry of the virus.

• Journal of Applied Biological Chemistry
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• 제61권3호
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• pp.267-273
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• 2018

기능성 천연물 소재로서의 가능성을 검토하고자 캐모마일 추출물의 미백 효과를 조사하고 melanin 생성 반응에 관여하는 물질 억제에 대한 기전을 규명하고자 하였다. 캐모마일을 water와 60% ethanol을 이용하여 추출하였고, 얻어진 추출물을 phenolic 농도별로 설정하여 tyrosinase 저해 활성을 알아보았을 때, water 추출물의 경우 효과가 미비하였고, 60% ethanol 추출물에서는 농도 의존적으로 저해 활성이 나타내어 melanin 생성 저해 효과가 있을 것으로 판단되었다. 캐모마일 60% ethanol 추출물을 동결건조하여 얻어진 분말을 이용하여 B16F10 melanoma cell에 대한 세포 독성을 측정 결과, $75{\mu}g/mL$의 농도에서부터 독성이 관찰되어 농도 구간을 10, 25, $50{\mu}g/mL$으로 선정하였다. ${\alpha}-MSH$로 자극한 B16F10 melanoma cell에서 melanin 생성량을 측정하여 캐모마일의 melanin 성성 억제 효능과 melanin 생성에 영향을 미치는 단백질인 tyrosinase, MITF, TRP-1, TRP-2의 단백질 발현 억제 효과를 알아본 결과, 캐모마일의 농도가 높아짐에 따라 농도 의존적으로 melanin 생성 함량이 감소하였고, tyrosinase, TRP-1, TRP-2, 등의 단백질 발현량 또한 감소하는 것을 확인할 수 있었다. 따라서 캐모마일 추출물은 B16F10 melanoma cell에서 melanin의 생성을 억제하고, melanin 생성 관련 단백질발현을 억제하는 효과가 있음을 확인하였다. 위의 결과들로 인하여 캐모마일은 미백 기능성 식품 산업화를 위한 유용한 자원으로 활용 될 것으로 예상되며, 추후 산업적 응용을 위한 지속적인 연구가 진행되어야 할 것으로 판단되었다.