[논문]섬괴불나무(Lonicera insularis Nakai) 추출물의 면역자극 및 항비만 활성

유주형; 여주호; 최민영; 이재원; 금나경; 안미연; 정진부

doi:10.7732/kjpr.2022.35.4.417

섬괴불나무(Lonicera insularis Nakai) 추출물의 면역자극 및 항비만 활성
Immunostimulatory and Anti-Obesity Activity of Lonicera insularis Nakai Extracts in Mouse Macrophages RAW264.7 Cells and Mouse Adipocytes 3T3-L1 Cells 원문보기

韓國資源植物學會誌 = Korean journal of plant resources, v.35 no.4, 2022년, pp.417 - 427

유주형 (국립안동대학교 생약자원학과) , 여주호 (국립안동대학교 생약자원학과) , 최민영 (국립안동대학교 생약자원학과) , 이재원 (국립안동대학교 생약자원학과) , 금나경 (국립안동대학교 생약자원학과) , 안미연 (국립산림품종관리센터) , 정진부 (국립안동대학교 생약자원학과)

초록
AI-Helper

본 연구에서는 섬괴불나무 열매(LIF), 잎(LIL) 그리고 줄기(LIS) 추출물의 면역증진 활성과 섬괴불나무 열매(LIF) 추출물의 항비만 활성을 평가하였다. 섬괴불나무 열매(LIF), 잎(LIL) 그리고 줄기(LIS) 추출물은 RAW264.7 세포에서 NO, iNOS, COX-2, IL-1𝛽, TNF-𝛼와 같은 면역증진인자의 생성을 증가시켰으며, IL-1𝛽의 발현은 NO생성과 관련된 것으로 보여진다. 면역증진인자은 TLR2/4를 통해 MAPKs중 p38 그리고 JNK를 자극하여 발현이 유도되는 것으로 판단된다. 항비만 실험에서, 섬괴불나무 열매(LIF) 추출물은 AMPK, HSL, ATGL의 발현 증가와 perilipin-1 발현 억제를통해 지질분해를 유도하여 세포 내 지질축적을 억제하는 것으로 나타났으며, 갈색지방세포로의 분화유도와 에너지 대사에 관여하는 인자인 PRDM16, PGC-1𝛼의 발현유도를 통해서도 지질축적을 억제하는 것으로 판단된다. 향후 섬괴불나무 추출물은 건강 보조제 및 기능성 식품으로의 활용이 가능할 것으로 판단되지만, 섬괴물나무 추출물의 어떠한 성분이 면역과 항비만 활성에 영향을 미치는지에 대한 성분분석이 필요하다. 또한, 본 연구는 세포를 이용한 실험으로 정확한 분석을 위해서는 동물모델을 이용한 섬괴불나무 추출물의 면역증진 및 항비만 활성에 관한 추가적인 연구가 진행되어야 할 것이다.

Abstract ▼ AI-Helper

In this study, we investigated in vitro immuno-stimulatory and anti-obesity activity of fruit (LIF), leaves (LIL) and stems (LIS) from Lonicera insularis Nakai in mouse macrophages RAW264.7 cells and mouse pre-adipocytes 3T3-L1 cells. LIF, LIL and LIS increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α) and activated phagocytosis in RAW264.7 cells. Inhibition of toll-like receptor 2/4 (TLR2/4) partly blocked LIF, LIL and LIS mediated production of immunostimulatory factors. In addition, inhibition of mitogen-activated protein kinases (MAPK) signaling attenuated the production of immunostimulatory factors induced by LIF, LIL and LIS. Based on these results of this study, LIF, LIL and LIS is thought to activate macrophages the production of immunostimulatory factors and phagocytosis through toll-like receptor 2/4 (TLR2/4) and MAPKs signaling pathway. In anti-obesity study, LIF reduced the lipid accumulation in 3T3-L1 cells. LIF increased the protein phosphorylation expressions such as AMP-activated protein kinase (AMPK), hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL) related to the lipolysis of the adipocytes. In addition, LIF increased the expression of proteins involved in energy metabolism and brown adipose tissues differentiation such as peroxisome proliferator-activated receptor gamma coativator 1α (PGC-1α) and PR domain-containing16 (PRDM16). These results suggest that LIF is involved in lipid accumulation inhibition through expressing the proteins such as lipolysis and differentiation of white adipocytes to brown adipocytes.

주제어

표/그림 (7)

표 Table 1. Sequence of oligonucleotide primers used for RT-PCR
그림 Fig. 1. Effect of LIF, LIL and LIS on macrophages activation in RAW264.7 cells. RAW264.7 cells were treated with LIF, LIL and LIS for 24 h. NO level (A), mRNA level (B). GAPDH was used as internal control for RT-PCR. *P < 0.05 compared to the cells without the treatment.
그림 Fig. 2. Effect of TLR2/4 on the production of immunostimulatory factors in LIF, LIL and LIS treated RAW264.7 cells. RAW264.7 cells were pre-treated with C29 (TLR2 inhibitor, 100 μM) or TAK-242 (TLR4 inhibitor, 5 μM) for 2 h and treated with LIF, LIL and LIS for 24 h. NO level (A) and mRNA level (B) were measured by Griess assay and RT-PCR, respectively. GAPDH was used as internal control for RT-PCR. *P < 0.05 compared to the cells without the treatment. ^#P < 0.05 compared to the cells treated with LIF, LIL and LIS alone.
그림 Fig. 3. Effect of MAPK signaling pathway on the production of immunostimulatory factors in LIF, LIL and LIS treated RAW264.7 cells. RAW264.7 cells were pre-treated with PD98059 (ERK1/2 inhibitor, 40 μM), SB203580 (p38 inhibitor, 40 μM), SP600125 (JNK inhibitor, 40 μM) for 2 h and then treated with LIF, LIL and LIS for 24 h. NO level (A) and mRNA level (B) were measured by Griess assay and RT-PCR. GAPDH and Actin were used as internal control for RT-PCR, Western blot. *P < 0.05 compared to the cells without the treatment. ^#P < 0.05 compared to the cells with LIF, LIL and LIS alone.
그림 Fig. 4. The Effect of LIF, LIL and LIS on inhibition of adipocyte differentitaion in 3T3-L1 cells. 3T3-L1 cells were co-treated with DMI/insulin and LIF, LIL and LIS for 8 Days. *P < 0.05 compared to the cells without the treatment. ^#P < 0.05 compared to the cells treated with DMI/insulin.
그림 Fig. 5. The Effect of LIF on lipolysis of adipocyte in 3T3-L1 cells. 3T3-L1 cells were co-treated with DMI/insulin and LIF for 8 Days. Actin was used as internal control for Western blot.
그림 Fig. 6. The Effect of LIF to Brown adipose tissue differentiation and energy metabolism and heat production induction protein expression in 3T3-L1 cells. 3T3-L1 cells were cotreated with DMI/insulin and LIF for 8 Days. Actin was used as internal control for Western blot.

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