H2O2 처리된 LLC-PK₁세포에서 Redox Status 및 NF-κB Signaling에 대한 夏枯草의 효과
Effects of Prunellae Herba on the H2O2-Treated LLC-PK₁ Cell's Redox Status and NF-κB Signaling
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This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O2-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE2) were measured in the H2O2-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O2-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H2O2-treated cell death dose-dependently. It reduced the generation of ·O2-, NO, ONOO- and PGE₂ in the H2O2-treated renal epitheial cells(LLC-PK1 cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.
Abstract
This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O2-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE2) were measured in the H2O2-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O2-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H2O2-treated cell death dose-dependently. It reduced the generation of ·O2-, NO, ONOO- and PGE₂ in the H2O2-treated renal epitheial cells(LLC-PK1 cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.
참고문헌 (34)
- Halliwell, B., "Free radicals in Biology and Medicine", pp.246~350 (1999)
- Ryu, S. N., "Antioxidative activity and varietal difference of cyanidin 3-glucoside and peonidin 3-glucoside contents in pigmented rice", Kor J. Crop Sci, 45(4), pp.257~260 (2000)
- Alfadda, A. A., "Reactive oxygen species in health and disease", J Biomed Biotechnol, 2012, pp.936486~ (2012)
- Valko, M., "Free radicals and antioxidants in normal physiological functions and human disease", Int J Biochem Cell Biol, 39, pp.44~84 (2007)
- 채성원, "NF - ΚB 활성화와 면역학적 기능", 대한이비인후과학회지 두경부외과학, 48(3), pp.284~288 (2005), 대한이비인후과학회
- Golan, D. E., "Principles of Pharmacology: The Pathophysiologic Basis of Drug Theraphy", pp.743~750 (2008), EPUBLIC
- 이명식, "내분비-대사질환에서 NF-κB의 최신 지견", Endocrinology and Metabolism, 21(5), pp.352~363 (2006), 대한내분비학회
- University of Korean Medicine Joint Publication Committee., "Bonchohak", pp.209~210 (2007), Yeongnimsa
- Seo, B. I., "The effects of Prunellae Spica on immune response and anti-allergic reaction", J. of Herbology, 12(1), pp.7~24 (1997)
- 장현주, "하고초물추출물이 LPS로 활성화된 Raw 264.7 cell에서의 염증매개물질 억제효과", 동의생리병리학회지, 23(3), pp.599~607 (2009), 한의병리학회
- 허자경, "免疫活性에 의한 夏枯草의 癌轉移 抑制 效果", 대한한방부인과학회지, 22(1), pp.41~52 (2009), 대한한방부인과학회
- Im, S. B. N., "Effects of Labiatae Herba Prunellae Aqua-acupuncture in Hyperthyroidism Rats", The Journal of Korean Oriental Medical Society, 18(1), pp.399~410 (1997)
- 허자경, "夏枯草에서 추출한 Ursolic acid의 파골세포 분화 억제 효과", 대한한방부인과학회지, 27(2), pp.59~70 (2014), 대한한방부인과학회[DOI]
- 차지혜, "하고초 열수추출물이 대식세포 면역만응에 미치는 영향", 대한한방부인과학회지, 23(3), pp.91~100 (2010), 대한한방부인과학회
- 김난슬, "하고초(Spica prunellae) 추출물의 항비만 및 산화적 스트레스 억제활성", 산업식품공학, 18(4), pp.413~418 (2014), 한국산업식품공학회[DOI]
- 홍은숙, "하고초 추출물의 항노화 효과에 관한 연구", 대한화장품학회지, 34(2), pp.129~135 (2008), 대한화장품학회
- Tada, H., "An improved colorimetric assay for interleukin 2", J Immunol Methods, 93(2), pp.157~165 (1986)
- Cathcart, R., "Detection of picomole levels of hydroperoxides using a fluorescent dichlorofluorescein fluorescent assay", Anal Biochem, 134, pp.111~116 (1983)
- Nagata, N., "Inhibitory effects of catecholamines and anti-oxidants on the fluorescence reaction of 4,5-diaminofluorescein, DAF-2, a novel indicator of nitric oxide", J Biochem Tokyo, 125, pp.658~661 (1999)
- Kooy, N. W., "Peroxynitrite-mediated oxidation of dihydrorhodamine 123", Free Radic Res Commun, 16, pp.149~156 (1994)
- Lowry, O. H., "Protein measurement with the Folin phenol reagent", J Biol Chem, 193(1), pp.265~275 (1951)
- Rubin, E., "Essentials of Rubin’s Pathology", pp.7~8 (2013), KOONJA
- Carr, A. C., "Oxidation of LDL by myeloperoxidase and reactive nitrogen species: reaction pathways and antioxidant protection", Arterioscler Thromb Vasc Biol, 20, pp.1716~1723 (2000)
- Chung, H. Y., "The inflammation hypothesis of aging: Molecular modulation by calorie restriction", Ann N Y Acad Sci, 928, pp.327~335 (2001)
- Kim, H. J., "The effect of age on cyclooxygenase-2 gene expression: NF-κB activation and IκBα degradation", Free Radic Biol Med, 28(5), pp.683~692 (2000)
- Crow, J. P., "Sensitivity of the essential zinc-thiolate moiety of yeast alchol dehydrogenase to hypochlorite and peroxynitrite", Biochemistry, 34(11), pp.3533~3552 (1995)
- DiDonato, J. A., "A cytokine-responsive IkB kinase that activates the transcriptional factor NF-kB", Nature, 388, pp.548~554 (1997)
- Lin, A., "NF-kB in cancer: a marked target", Semin Cancer Biol, 13, pp.107~114 (2003)
- Li, Q., "Severe liver degeneration in mice lacking the IkappaB kinase 2 gene", Science, 284, pp.321~325 (1999)
- Greten, F. R., "IKKbeta links inflammation and tumorigenesis in a mouse model of colitis-associated cancer", Cell, 118, pp.285~296 (2004)
- Duyao, M. P., "Transactivation of the c-myc promoter by human T cell leukemia virus type 1 tax is mediated by NF-κB", J Biol Chem, 267, pp.16288~16291 (1992)
- Chung, H. Y., "Molecular inflammation hypothesis of aging based on the anti-aging mechanism of calorie restriction", Micro Res Techinoq, 59, pp.264~272 (2002)
- Baeuerle, P. A., "Function and activation of NF-κB in the immune system", Annu Rev Immunol, 12, pp.141~179 (1994)
- Osborn, L., "Direct Expression Cloning of Vascular Cell Adhesion Molecule 1, a Cytokine-Induced Endothelial Protein That Binds to Lymphocytes", Cell, 59, pp.1203~1211 (1989)
