고구마 소주 주박에 의한 RAW 264.7 세포주에서 lipopolysaccharide로 유도된 염증 반응의 억제 효과

국문 초록 (Abstract)

본 연구에서는 고구마 소주로부터 획득한 에탄올 추출물 및 순차적 유기용매 분획물을 제조하고, 이들 시료가 LPS로 활성화된 마우스 대식세포 RAW 264.7 세포주에서 nitric oxide (NO) 생산과 세포생존율에 미치는 영향을 연구하였다. 처리한 시료 중 ethyl acetate 분획물(KSD-E8-3) 처리군에서 LPS로 염증 유도된 RAW 264.7 세포주의 생존율에는 영향 없이 NO 생성을 현저하게 감소 시킴을 확인하였다. 또한 분획물 KSD-E8-3 처리에 의해서 염증매개 인자인 COX-2, iNOS 그리고 TNF-alpha의 유전자의 발현이 감소되었으며, iNOS 단백질의 발현이 현저히 감소됨을 Western blot으로 확인하였다. 또한, 분획물 KSD-E8-3 처리에 의해 세포내 대표적인 신호 전달 단백질인 3가지의 MAPK 단백질(p38, JNK, ERK1/2)의 인산화가 모두 억제됨을 확인하였다. 종합적으로, 이러한 연구결과는 고구마 소주 주박에 함유된 생리활성 물질에 의한 항염증 활성 및 작용 기전을 이해하는데 도움을 줄 것으로 기대된다.

다국어 초록 (Multilingual Abstract)

In the current study, the ethanol extracts and their subsequent organic solvent fractions from lees of sweet potato soju were prepared and the prepared samples were designated as from KSD-E8-1 to KSD-E8-5. Their effects on cell viability and nitric oxide (NO) production in mouse macrophage RAW 264.7 cells were investigated. The results showed that the ethyl acetate fraction (KSD-E8-3) of lees extracts from sweet potato soju significantly decreased nitric oxide (NO) production in LPS-activated RAW 264.7 cells, whereas they did not affect cell viabilities. The fraction KSD-E8-3 reduced the expression of pro-inflammatory genes such as COX-2, iNOS and TNF-alpha and also decreased protein expression of iNOS in a dose dependent manner, which were detected with RT-PCR and Western blot analysis, respectively. In addition, we detected the expression of mitogen-activated protein kinases (MAPKs) such as p38, JNK, and ERK1/2 and their phosphorylated forms. The results indicated that the treatment of the fraction KSD-E8-3 did inhibit phosphorylation of p38, JNK, and ERK1/2 MAPKs, indicating that the fraction KSD-E8-3 regulates LPS-induced inflammatory response via suppressing MAPK signaling pathway. Overall, these results may contribute to understand the molecular mechanism of anti-inflammatory effects by the ethyl acetate fraction of lees extracts from sweet potato soju.

목차 (Table of Contents)

• 서론
• 재료 및 방법
• 결과 및 고찰
• References
• 초록

참고문헌 (Reference)

1 김태영, "주박 추출물의 항균활성,항산화 및 SOD 유사활성 효과" 한국식품영양학회 23 (23): 299-305, 2010

2 박미정, "주박 추출물과 이들의 유기용매 분획물에 의한 항염증 활성" 한국생명과학회 24 (24): 843-850, 2014

3 김미선, "전통주 주박의 항혈전 활성 평가" 한국생명과학회 24 (24): 865-872, 2014

4 강형택, "대장암 세포주에서 주박 추출물의 유기용매 분획물의 항성장 활성" 한국생명과학회 24 (24): 967-972, 2014

5 Kim, H. G., "p38αMAPK is required for arsenic-induced cell transformation" 22331-, 2015

6 Akira, S., "Toll-like receptor signaling" 4 : 499-511, 2004

7 Eisinger, A. L., "The role of cyclooxygenase-2 and prostaglandins in colon cancer" 82 : 147-154, 2007

8 Vlkova, M., "Regulatory B cells in CVID patients fail to suppress multifunctional IFN-gamma+TNF-alpha+CD4+ T cells differentiation" 29 : 292-300, 2015

9 Li, G., "Regulatory B cell function is suprresed by smoking and obesity in H. pylori-infected subjects and is correlated with elevated risk of gastric cancer" 10 : e0134591-, 2015

10 Anderson, N., "Molecular mechanisms and therapeutic targets in steatosis and steatoheptitis" 60 : 311-357, 2008

11 Ju, A., "Methanol extracts of Xanthium sibiricum roots inhibit inflammatory response via the inhibition of nuclear factor-κB (NF-κB) and signal transducer and activator of transctiption 3 (STAT3) in murine macrophage" 2015

12 Monick, M. M., "Lipopolysaccharide activates Akt in human alveolar macrophages resulting in nuclear accumulation and transcriptional activity of beta-catenin" 166 : 4713-4720, 2001

13 Kojda, G., "Interactions between NO and reactive oxygen speices : pathophysiological importance in atherosclerosis, hypertension, diabetes and heart failure" 43 : 562-571, 1999

14 Kindt, T. J., "Innate immunity : Tenneys. Kuby Immunology" Freeman Press 52-73, 2007

15 Lee, E. S., "Inhibition of nitric oxide and tumor necrosis factor-alpha (TNF-alpha) production by prepenone compound through blockade of nuclear factor (NF)-κB activation in cultured murine macrophages" 27 : 617-620, 2004

16 Xu, C., "Hyaluronan ameliorates LPS-induced acute lung injury in mice via Toll-like receptor (TLR) 4-dependent signaling pathway" 28 : 1050-1058, 2015

17 Lo, J. Y., "Curcumenol isolated from Curuma zedoaria suppresses Akt-mediated NF-κB activation and p38MAPK signaling pathway in LPS-stimulated BV-2 microlial cells" 6 : 3550-3559, 2015

18 Sahu, B. D., "Baicalein, a bioflavonoid, prevents cisplatin-induced acute kidney injury by up-regulating antioxidant defense and down-regulating the MAPKs and NF-κB pathways" 10 : e0134139-, 2015

19 Jin, J. H., "Anti-inflammatory and anti-arthritic activity of the roots of sophora flavescens" 127 : 589-595, 2010

20 Chelsky, Z. L., "A resveratrol analogue promotes ERK MAPK-dependent STAT3 serine and tyrosine phosphorylation alterations and antitumor effects in vitro against human tumor cells" 88 : 524-533, 2015

21 Jacobsen, M. C., "A critical role for ATF2 transcription factor in the regulation of E-selectin expression in response to non-endotoxin components of Neisseria meningitides" 2015

22 Syed Hussein, S. S., "(+)-Catechin attenuates NF-κB activation through regulation of Akt, MAPK, and AMPK signaling pathways in LPS-induced BV-2 microglial cells" 43 : 927-952, 2015