LPS로 염증 유도된 RAW 264.7세포에 대한 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물의 항염증 효과

국문 초록 (Abstract)

본 연구에서는 참콩풍뎅이(Popillia flavosellata) 에탄올 추출물(PFE)의 항염증 효능을 분석하기 위해 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 대식세포인 RAW 264.7에 처리 시 최고 처리농도인 2,000 ㎍/ml까지 통계적인 유의성 있는 독성이 없음을 확인하였다. LPS (100 ng/ml)로 염증 유도된 RAW 264.7 세포에 PFE를 농도별(500, 1,000, 2,000 ㎍/ml)로 동시 처리 시 농도 의존적으로 염증성사이토카인인 TNF-α와 IL-6의 단백질 생성을 통계적인 유의성(p＜0.001)있게 억제함을 확인하였다. 또한 염증 유도된 RAW 264.7 세포에 PFE 동시 처리 시 NF-κB p65의 핵으로 이동이 차단됨과 iNOS와 COX-2의 단백질 발현을 감소시키는 것을 확인하였다. 이상의 연구결과를 통해 참콩풍뎅이는 염증에 의해 활성화된 TLR-4 신호전달과정을 조절하는 NF-κB p65의 활성과 염증성사이토카인 TNF-α와 IL-6의 생성 및 염증성효소 iNOS와 COX-2의 생성을 억제하는 항염증 효능이 있음을 확인하였다.

다국어 초록 (Multilingual Abstract)

The beetle Popillia flavosellata has been no reported its functional effects. In this study, we investigated the anti-inflammatory effect of P. flavosellata ethanol extract (PFE) on RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for the induction of inflammation. First, we examined the cytotoxicity of PFE in the RAW 264.7 cells at a concentration of 2,000 μg/ml or less. To evaluate the anti-inflammatory effects of PFE, we investigated the expression levels of proinflammatory cytokines, such as tumor necrosis factor (TNF)-α and interleukin (IL)-6, and proinflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether PFE inhibited the translocation of nuclear factor kappa B (NF-κB) p65 into the nucleus in the LPS-induced RAW 264.7 cells. We found that the protein levels of TNF-α and IL-6 were decreased in the LPS-induced RAW 264.7 cells after the treatment with PFE in a dose-dependent manner. In addition, we confirmed that PFE inhibited the translocation of NF-κB p65 into the nucleus, as well as the protein expression levels of iNOS and COX-2. Accordingly, we propose that PFE exerts an anti-inflammatory effect through the down-regulation of NF-κB p65, TNF-α, IL-6, iNOS, and COX-2 via the toll like receptor (TLR)-4 inflammatory signaling pathway.

목차 (Table of Contents)

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

참고문헌 (Reference)

1 Mi Yeon Chung, "흰점박이꽃무지(Protaetia brevitarsis)의 일반성분 및 유해물질 분석" 한국생명과학회 23 (23): 664-668, 2013

2 이상한, "메뚜기류 추출물의 염증 조절작용 및 세포사멸보호 효과" 한국식품저장유통학회 13 (13): 796-802, 2006

3 유정미, "국산 및 중국산 갈색거저리(Tenebrio molitor)의 영양성분 및 유해물질 비교분석" 한국식품영양과학회 42 (42): 249-254, 2013

4 Tracey, D., "Tumor necrosis factor antagonist mechanisms of action : a comprehensive review" 117 : 244-279, 2008

5 Ahmadi-Ashtiani, H. R., "The inhibitory effect of silymarin on cell viability and cellular COX-2 and iNOS level in HepG2 cell line" 7 : 2008-2014, 2013

6 Baldwin, A. S. Jr, "The NF-kappa B and I kappa B proteins : new discoveries and insights" 14 : 649-683, 1996

7 Cerella, C., "Targeting COX-2 expression by natural compounds : a promising alternative strategy to synthetic COX-2 inhibitors for cancer chemoprevention and therapy" 15 : 1801-1815, 2010

8 Mitchell, J. A., "Selectivity of nonsteroid anti-inflammatory drugs as inhibitors of constitutive and inducible cyclo-oxygenase" 90 : 11693-11697, 1993

9 Howes, L. G, "Selective COX-2 inhibitors, NSAIDs and cardiovascular events-is celecoxib the safest choice?" 3 : 831-845, 2007

10 Lee, S. Y., "SHINBARO, a new herbal medicine with multifunctional mechanism for joint disease : first therapeutic application for the treatment of osteoarthritis" 34 : 1773-1777, 2011

11 Rajendram, R., "Photoreceptor mitochondrial oxidative stress in early experimental autoimmune uveoretinitis" 91 : 531-537, 2007

12 Medzhitov, R, "Origin and physiological roles of inflammation" 454 : 428-435, 2008

13 Miyataka, M., "Nitric oxide, anti-inflammatory drugs on renal prostaglandins and cyclooxygenase-2" 39 : 785-789, 2002

14 Salvemini, D., "Nitric oxide activates cyclooxygenase enzymes" 90 : 7240-7244, 1993

15 Giuliani, C., "NF-κB transcription factor : role in the pathogenesis of inflammatory, autoimmune, and neoplastic diseases and therapy implications" 152 : 249-253, 2001

16 Tak, P. P., "NF-kappaB : a key role in inflammatory diseases" 107 : 7-11, 2001

17 Nakanishi, M., "Multifaceted roles of PGE2 in inflammation and cancer" 35 : 123-137, 2013

18 윤영일, "LPS로 유도된 RAW 264.7세포에 대한 벼메뚜기(Oxya chinensis sinuosa) 에탄올 추출물의 항염증 효과" 한국생명과학회 24 (24): 370-376, 2014

19 Kim, J. J., "Investigation of host plants and seasonal occurrence of Popillia flavosellata(Coleoptera : Rutelidae)in golf courses" 25 : 22-29, 2011

20 Gabay, C, "Interleukin-6 and chronic inflammation" 8 : S3-, 2006

21 Brennan, F. M., "Inhibitory effect of TNF alpha antibodies on synovial cell interleukin-1 production in rheumatoid arthritis" 2 : 244-247, 1989

22 Vane, J. R, "Inhibition of prostaglandin synthesis as a mechanism of action for the aspirin-like drugs" 231 : 232-235, 1971

23 Kang, C. H., "Inhibition of lipopolysaccharide-induced iNOS, COX-2, and TNF-α expression by aqueous extract of Orixa Japonica in RAW 264. 7 cells via suppression of NF-κB activity" 10 : 161-168, 2011

24 Iwalewa, E. O., "Inflammation : the foundation of diseases and disorders. A review of phytomedicines of south african origin used to treat pain and inflammatory conditions" 6 : 2868-2885, 2007

25 Wang, X., "Effect of osteoprotegerin in combination with interleukin-6 on inhibition of osteoclast differentiation" 16 : 277-280, 2013

26 Van Huis, A., "Edible insects: future prospects for food and feed security" Food and agriculture Organization of the United Nations 2013

27 Shafran, Y., "Correlative analyses of nitric oxide generation rates and nitric oxide synthase levels in individual cells using a modular cell-retaining device" 84 : 7315-7322, 2012

28 Swierkosz, T. A., "Co-induction of nitric oxide synthase and cyclo-oxygenase : interactions between nitric oxide and prostanoids" 114 : 1335-1342, 1995

29 Ferrero-Miliani, L., "Chronic inflammation : Importance of NOD2 and NALP3 in interleukin-1beta generation" 147 : 227-235, 2007

30 Rao, C. V., "Chemopreventive properties of a selective inducible nitric oxide synthase inhibitor in colon carcinogenesis, administered alone or in combination with celecoxib, a selective cyclooxygenase-2 inhibitor" 62 : 165-170, 2002

31 Paek, M. K., "Checklist of Korean insects" Nature & Ecology 113-, 2010

32 Monaco, C., "Canonical pathway of nuclear factor kappa B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis" 101 : 5634-5639, 2004

33 Hink, U., "COX-2, another important player in the nitric oxide-endothelin cross-talk: good news for COX-2 inhibitors?" 98 : 1344-1346, 2006

34 Arora, R. B., "Anti-inflammatory studies on Curcuma longa(turmeric)" 59 : 1289-1295, 1971

35 Vlassara, H, "Advanced glycation in health and disease role of the modern environment" 1043 : 452-460, 2005