[논문]3T3-L1 전구지방세포에서 개구리자리(Ranunculus sceleratus) 추출물의 AMPK 신호전달을 통한 지방생성 억제 효과

김예지; 조성필; 이희주; 홍금란; 김경현; 류시윤; 정주영

doi:10.15429/jkomor.2022.22.1.30

[국내논문] 3T3-L1 전구지방세포에서 개구리자리(Ranunculus sceleratus) 추출물의 AMPK 신호전달을 통한 지방생성 억제 효과
Extract of Ranunculus sceleratus Reduced Adipogenesis by Inhibiting AMPK Pathway in 3T3-L1 Preadipocytes 원문보기

한방비만학회지 = Journal of Korean Medicine for Obesity Research, v.22 no.1, 2022년, pp.30 - 37

김예지 (충남대학교 수의과대학 조직학교실) , 조성필 (충남대학교 수의과대학 조직학교실) , 이희주 (충남대학교 수의과대학 조직학교실) , 홍금란 (충남대학교 수의과대학 조직학교실) , 김경현 (충남대학교 수의과대학 조직학교실) , 류시윤 (충남대학교 수의과대학 해부학교실) , 정주영 (충남대학교 수의과대학 조직학교실)

Abstract ▼ AI-Helper

Objectives: Adipogenesis is the process by which pre-adipocytes are differentiated into adipocytes. It also plays an important role in adipocyte formation and lipid accumulation. Ranunculus sceleratus (R. sceleratus) extracts are used for the treatment of various diseases such as hepatitis, jaundice, and tuberous lymphadenitis in oriental medicine. However, its effect on adipogenesis has not yet been studied. In this study, we investigated the effects of R. sceleratus on adipogenesis in 3T3-L1 cells. Methods: Cells were treated with 50, 100, and 200 ㎍/ml of R. sceleratus and cell viability was evaluated. To differentiate the 3T3-L1 preadipocytes, a 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI) solution were used. The accumulation of lipid droplets was determined by Oil Red O staining. The expression levels of adipogenesis-related proteins were also determined. Results: MDI solution differentiated the preadipocytes into adipocytes and accumulation of lipids was observed in the differentiated 3T3-L1 cells. Interestingly, the amount of lipid droplets was reduced after R. sceleratus treatment. In addition, the expression levels of key adipogenic transcription factors, such as CCAAT/enhancer-binding proteins-𝛼 (C/EBP-𝛼) and peroxisome proliferator-activated receptors-𝛾 (PPAR-𝛾) were also reduced after R. sceleratus treatment. Furthermore, R. sceleratus increased AMP-activated kinase (AMPK) phosphorylation and decreased sterol regulatory element-binding protein-1 expression. Conclusions: Our results showed that R. sceleratus reduced preadipocyte differentiation by inhibiting C/EBP-𝛼 and PPAR-𝛾 levels via the AMPK pathway. Therefore, we suggest that R. sceleratus may be potentially used as an anti-adipogenic agent.

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표/그림 (5)

그림 Fig. 1. Effect of R. sceleratus extract on the cell viability of 3T3-L1 preadipocytes. 3T3-L1 preadipocytes were treated with various concentrations (50~200 μg/ml) of R. sceleratus (A) Cell viability of 3T3-L1 preadipocytes after 24 h. (B) Cell viability of 3T3-L1 preadipocytes after 48 h. (C) Cell viability of 3T3-L1 preadipocytes after 72 h.
그림 Fig. 2. Effect of R. sceleratus extract on the differentiation of 3T3-L1 adipocytes via Oil Red O staining. 3T3-L1 preadipocytes were treated with various concentrations (50~200 μg/ml) of R. sceleratus. (A) Cell culture and differentiation protocol. (B) Intercellular lipid droplets were stained with Oil Red O. After 10 days differentiation, representative phasecontrast photomicrographs at 200x and 400x magnification depictied of 3T3-L1 adipocyte. Control (-) include a and b, control (+) include b and g, 50 μg/ml, include c and h, 100 μg/ml include d and i, 200 μg/ml include e and j. (C) Quantification of lipid accumulation by eluting with isopropanol. The Data were presented as mean±standard error of the mean from three independent experiments. Con (-): control without MDI media, Con (+): control with MDI media, MDI: 0.5 mM IBMX, 1 μM DEX, 10 μg/ml insulin, R. sceleratus: Ranunculus sceleratus, IBMX: 3-isobutyl-1-methylxanthine, DEX: dexamethasone, FBS: fetal bovine serum. #P<0.05 compared with the control (-); *P<0.05, **P<0.01 compared with the control (+).
그림 Fig. 3. Effect of R. sceleratus on the expression levels of the C/EBP-α and PPAR-γ in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with various concentrations (50~200 μg/ml) of R. sceleratus. GAPDH was used as a loading control. The Data were presented as mean±standard error of the mean from three independent experiments. PPAR-γ: peroxisome proliferator-activated receptor-γ, C/EBP-α: CCAAT/enhancer-binding proteins-α, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, C (-): control (-), C (+): control (+). ^##P<0.01 compared with the control (-); **P<0.01 compared with the control (+).
그림 Fig. 4. Effect of R. sceleratus on the expression of AMPK/SREBP-1 signaling pathway in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with various concentrations (50~200 μg/ml) of R. sceleratus. GAPDH was used as a loading control. The Data were presented as mean±standard error of the mean from three independent experiments. AMPK: AMP-activated protein kinase, p-AMPK: phosphorylated AMPK, SREBP-1: sterol regulatory element-binding protein-1, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, C (-): control (-), C (+): control (+). ^##P<0.01 compared with the control (-); *P<0.05,**P<0.01 compared with the control (+).
그림 Fig. 5. AMPK signaling pathway of adipogenesis inhibition of R. sceleratus in 3T3-L1. PPAR-γ: peroxisome proliferator-activated receptor-γ, C/EBP-α: CCAAT/enhancer-binding proteins-α, AMPK: AMP-activated protein kinase, SREBP-1: sterol regulatory element-binding protein-1.

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