초록 ▼

Ⅳ. 연구개발결과
○ 인플라마솜 활성 조절 소재 탐색 실험법 확립
- Reporter cell line을 이용한 인플라마솜 활성조절 소재 탐색방법 확립 및 검증

○ 지유에 의한 인플라마솜 억제 및 작용기전 연구
- 지유(오이풀)에 의한 인플라마솜 활성화 억제를 다양한 인플라마솜 inducer를 사용하여 검증함.
- 지유(오이풀)가 LPS에 의한 septic shock을 예방함을 증명함
- 지유(오이풀)가 AOM유도 대장암 전암병변과 AOM/DSS 유도 대장암에 대한 예방 효과 가능성 제시

Ⅳ. 연구개발결과
○ 인플라마솜 활성 조절 소재 탐색 실험법 확립
- Reporter cell line을 이용한 인플라마솜 활성조절 소재 탐색방법 확립 및 검증

○ 지유에 의한 인플라마솜 억제 및 작용기전 연구
- 지유(오이풀)에 의한 인플라마솜 활성화 억제를 다양한 인플라마솜 inducer를 사용하여 검증함.
- 지유(오이풀)가 LPS에 의한 septic shock을 예방함을 증명함
- 지유(오이풀)가 AOM유도 대장암 전암병변과 AOM/DSS 유도 대장암에 대한 예방 효과 가능성 제시

○ 뚱딴지에 의한 인플라마솜 억제 및 작용기전 연구
- 뚱딴지에 의한 인플라마솜 활성화 억제를 다양한 인플라마솜 inducer를 사용하여 검증함.
- 뚱딴지가 AOM/DSS에 대한 예방 효과 가능성 제시

○ 고구마에 의한 인플라마솜 활성화 유도 및 작용기전 연구
- 고구마에 의한 인플라마솜 활성화 및 기전 연구
- 고구마가 AOM 유도 전암병변에 대한 예방 효과 가능성 제시

○ 땅두릅에 의한 면역세포 활성화 기전 및 세균감염에 대한 저항성 증강 검증
- 인플라마솜 경로를 활성화 시키거나 억제하지는 않으나 Raw264.7세포주와 BMDM 세포에서 MyD88 의존적 경로로 면역반응을 활성화 시킴.
- In vivo 실험을 통해 땅두릅이 Listeria 감염을 예방할 수 있음을 확인함.

○ 말굽버섯에서 분리한 fomentariol과 FF-8에 의한 항염증 활성 규명
- 말굽버섯 추출물이 항염 활성이 있다는 보고 있었으나 활성 성분은 잘 알려지지 않음.
- formentariol과 FF-8이 LPS에 의해 유도된 염증반응을 억제함을 확인함.

Abstract ▼

Inflammasome is a cytosolic multi-protein complex of nucleotide-binding domain and leucine rich repeat containing protein (NLRs) or AIM2, ASC and caspase-1 that are assembled in response to the detection of endogenous and pathogen-associated danger signals (1). Upon activation, cleaved caspase-1 act

Inflammasome is a cytosolic multi-protein complex of nucleotide-binding domain and leucine rich repeat containing protein (NLRs) or AIM2, ASC and caspase-1 that are assembled in response to the detection of endogenous and pathogen-associated danger signals (1). Upon activation, cleaved caspase-1 activates IL-1β and induces IL-1β secretion. Inflammsomes play a central role in innate immune responses and is essential in initiating adaptive immune responses against a wide array of pathogens (2-4). However, dysregulated production of IL-1β through inflammasome activation is associated with a number of inflammatory and metabolic disorders (5). Therefore targeting inflammsomes could be a promising therapeutic strategy to control various diseases. Here, we set up the protocol to screen materials extracted from agricultural/food resources for regulating inflammasome activation. Instead of ELISA method to detect IL-1β, we used IL-1β reporter cell line to screen over 150 extract collections. Selected extracts were further confirmed using mouse bone marrow derived macrophages (BMDM), IL-1β ELISA, immunoblotting cleaved IL-1β and cleaved caspase-1, and ASC pyroptosome formation. Furthermore, animal diseases models were applied to evaluate physiological activities in in vivo.
Our study indicated that hot water extract of Sanguisorba officinalis (HSO) effectively suppressed inflammasome activation triggered by ATP, nigericin, microbial pathogens, and dsDNA in bone marrow derived macrophages (BMDM). HSO significantly suppressed IL-1 β production in a dose dependent manner, which was well correlated with low levels of caspse-1 and ASC pyroptosome formation in HSO treated cells. This anti-inflammatory activity of HSO was further confirmed in mouse model of endotoxin induced septic shock. Oral administration of HSO reduced the level of IL-1β in serum and peritoneal cavity, leading to increased survival rate. Taken together, our results suggest that HSO is an effective inhibition inflammasome activation through NLRP3, NLRC4 and AIM2, and possibly would be useful for cure of inflammasome-mediated diseases. In addition, possible preventive effects of HSO on azoxymethan (AOM) induced preneoplastic lesion in colon and AOM/DSS induced inflammatory colon cancer were studied in mice model.
Aralia cordata (AC) is a known pain reliever and anti-inflammatory drug used in traditional Asian medicine. We found that HAC activated bone marrow-derived macrophages (BMDM) and increased levels of nitric oxide and proinflammatory cytokines in a dose-dependent manner. In addition, HAC induced the phosphorylation of NF-κB and mitogen-activated protein kinases (MAPKs), including JNK, ERK, and p38. Interestingly, these effects were absent in BMDM prepared from MyD88-knockout mice. Polysaccharides from HAC exerted stronger immunostimulatory effects compared to HAC itself. Furthermore, orally administered HAC clearly enhanced the clearance of the intracellular pathogen Listeria monocytogenes by boosting innate immune responses. These results demonstrate that HAC exerts. In addition, a possible preventive effect of HAC on AOM/DSS induced inflammatory colon cancer was investigated using mice model.
Ipomoea batatas has been known to possess anti-oxidant, ant-diabetic and anti-cancer activity. However, the effects of Ipomoea batatas leaf and stem extract on inflammasomes have not been studied yet. The NLRP3 inflammasome is a multiprotein signaling complex which consists of NLRP3 sensor, capase-1 and ASC. NLRP3 inflammsomes play an essential role in innate immune responses against microbial infection through maturation of IL-1ß and IL-18. Here, we investigated the effect of methanol extract of Ipomoea batatas (MEIB) on NLRP3 inflammasome using human monocytic cell line, THP-1. When THP-1 cells were treated with MEIB in the present of LPS, IL-1ß secretion triggered by LPS was intensified in dose-dependent manner. In addition, similar results were obtained when LPS-primed THP-1 cells were treated with MEIB. We also found that MEIB enhanced the cleavage of proIL-1β via caspase-1 activation in LPS-primed THP-1 cells. Using specific inhibitors, we found that MEIB stimulated THP-1 cells to produce IL-1ß through ATP and K+ efflux. Taken together, our results suggest that MEIB could be a NLRP3 inflammsome inducer and further studies are on-going to identify molecular mechanisms. In addition a ossible preventive effect of HSO on azoxymethan (AOM) induced preneoplastic lesion in colon was evaluated in mice model.
We also investigated the effects of methanol extract of Helianthus tuberosus (MEHT) on NLRP3 inflammasome in bone-marrow derived macrophage (BMDM). Our results demonstrated that MEHT effectively suppressed NLPR3 inflammasome activation in LPS primed and ATP or nigericine treated BMDM, resulting in reduced IL-1β secretion. This effect was well correlated with lower levels of caspase-1 in MEHT treated BMDM. Furthermore, MEHT significantly attenuated IL-1 β secretion mediated by AIM2 and NLRC4 inflammasome, which are induced by Listeria monocytogens and Salmonella typhimurium, respectively. These results indicate that MEHT may act as a broad spectrum inflammasome inhibitor. In addition, a possible preventive effect of HAC on AOM/DSS induced inflammatory colon cancer was investigated using mice model.Further study will test whether MEHT suppress inflammasome activation in vivo models.