본 연구에서는 상백피의 소화효소 저해활성과 3T3-L1 전지방세포의 분화 억제능을 기반으로 항비만 효능소재로서의 활용가능성을 평가하였다. 상백피의 에탄올 추출물(MRE)은 ${\alpha}$-amylase와 ${\alpha}$-glucosidase, pancreatic lipase를 활용한 소화효소 저해활성 평가 실험에서 각각 $7.86{\pm}0.36$, $0.12{\pm}0.03$, $7.93{\pm}0.11mg/mL$의 $IC_{50}$ 값을 보이며 우수한 억제 활성을 나타냈다. 또한 3T3-L1 전지방세포를 활용한 세포분화억제효능실험에서 MRE 처리군의 세포내 지방 축적율은 농도 의존적으로 감소되었다. 상백피의 항비만 작용 기전을 구명하기 위하여 adipogenesis 및 lipogenesis와 관련된 유전자 발현양상을 분석한 결과, 상백피 추출물 처리군에서는 생체내 지방대사 조절에 중요한 역할을 하는 FAS와 ACC 뿐 아니라 adipogenesis와 lipogenesis와 관련된 주요 전사요소인 $PPAR{\gamma}$와 $C/EBP{\alpha}$, SREBP-1c의 유전자 발현이 현저하게 억제되었다. qRT-PCR 분석 결과, 상백피 추출물의 anti-adipogenesis 효능은 전사단계에서의 관련 유전자 발현억제에 기인한다고 판단되었다. 본 실험결과 상백피 추출물은 전지방세포의 분화와 세포내 지질합성을 저해하고 비만과 관련 된 소화효소에 대한 저해활성을 나타내었다. 이러한 결과를 기반으로 상백피의 비만 예방 소재로서의 잠재적인 가능성을 확인하였다.
본 연구에서는 상백피의 소화효소 저해활성과 3T3-L1 전지방세포의 분화 억제능을 기반으로 항비만 효능소재로서의 활용가능성을 평가하였다. 상백피의 에탄올 추출물(MRE)은 ${\alpha}$-amylase와 ${\alpha}$-glucosidase, pancreatic lipase를 활용한 소화효소 저해활성 평가 실험에서 각각 $7.86{\pm}0.36$, $0.12{\pm}0.03$, $7.93{\pm}0.11mg/mL$의 $IC_{50}$ 값을 보이며 우수한 억제 활성을 나타냈다. 또한 3T3-L1 전지방세포를 활용한 세포분화억제효능실험에서 MRE 처리군의 세포내 지방 축적율은 농도 의존적으로 감소되었다. 상백피의 항비만 작용 기전을 구명하기 위하여 adipogenesis 및 lipogenesis와 관련된 유전자 발현양상을 분석한 결과, 상백피 추출물 처리군에서는 생체내 지방대사 조절에 중요한 역할을 하는 FAS와 ACC 뿐 아니라 adipogenesis와 lipogenesis와 관련된 주요 전사요소인 $PPAR{\gamma}$와 $C/EBP{\alpha}$, SREBP-1c의 유전자 발현이 현저하게 억제되었다. qRT-PCR 분석 결과, 상백피 추출물의 anti-adipogenesis 효능은 전사단계에서의 관련 유전자 발현억제에 기인한다고 판단되었다. 본 실험결과 상백피 추출물은 전지방세포의 분화와 세포내 지질합성을 저해하고 비만과 관련 된 소화효소에 대한 저해활성을 나타내었다. 이러한 결과를 기반으로 상백피의 비만 예방 소재로서의 잠재적인 가능성을 확인하였다.
Anti-obese effects of mulberry (Morus alba L.) root bark was investigated in vitro by measuring its inhibitory effect against 3T3-L1 preadipocyte differentiation and digestive enzymes such as ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase. Ethanol extract of m...
Anti-obese effects of mulberry (Morus alba L.) root bark was investigated in vitro by measuring its inhibitory effect against 3T3-L1 preadipocyte differentiation and digestive enzymes such as ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase. Ethanol extract of mulberry root bark (MRE) showed the potent inhibitory activities on ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase with $IC_{50}$ values of $7.86{\pm}0.36$, $0.12{\pm}0.03$ and $7.93{\pm}0.11mg/mL$, respectively. Furthermore, MRE significantly suppressed cellular lipid accumulation in 3T3-L1 cells in a dose-dependent manner. To elucidate the mechanism of MRE, we performed qRT-PCR and Western blotting for the expression of genes related with adipogenesis and lipogenesis. Treatment of MRE markedly suppressed the protein expression of $PPAR{\gamma}$, $C/EBP{\alpha}$ and SREBP-1c, as well as FAS and ACC, which are the key transcription factors and metabolic enzymes in adipogenesis and lipogenesis. In addition, qRT-PCR analysis indicated that the anti-adipogenesis effect of MRE might be due to its inhibition at transcription levels. These results demonstrate that MRE can effectively suppress adipocyte differentiation and inhibit key enzymes related to obesity. Our findings suggest that mulberry root bark may have a potential benefit in preventing obesity.
Anti-obese effects of mulberry (Morus alba L.) root bark was investigated in vitro by measuring its inhibitory effect against 3T3-L1 preadipocyte differentiation and digestive enzymes such as ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase. Ethanol extract of mulberry root bark (MRE) showed the potent inhibitory activities on ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase with $IC_{50}$ values of $7.86{\pm}0.36$, $0.12{\pm}0.03$ and $7.93{\pm}0.11mg/mL$, respectively. Furthermore, MRE significantly suppressed cellular lipid accumulation in 3T3-L1 cells in a dose-dependent manner. To elucidate the mechanism of MRE, we performed qRT-PCR and Western blotting for the expression of genes related with adipogenesis and lipogenesis. Treatment of MRE markedly suppressed the protein expression of $PPAR{\gamma}$, $C/EBP{\alpha}$ and SREBP-1c, as well as FAS and ACC, which are the key transcription factors and metabolic enzymes in adipogenesis and lipogenesis. In addition, qRT-PCR analysis indicated that the anti-adipogenesis effect of MRE might be due to its inhibition at transcription levels. These results demonstrate that MRE can effectively suppress adipocyte differentiation and inhibit key enzymes related to obesity. Our findings suggest that mulberry root bark may have a potential benefit in preventing obesity.
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문제 정의
This study was aimed to investigate the inhibitory effects of mulberry root bark on digestive enzymes and 3T3-L1 preadipocytes differentiation. MRE was further evaluated for the potential mechanism underlying the regulation of cell differentiation, in which the gene expression patterns involved in adipogenesis and lipogenesis of 3T3-L1 adipocyte cells were analyzed.
본 연구에서는 상백피의 소화효소 저해활성과 3T3-L1전지방세포의 분화 억제능을 기반으로 항비만 효능소재로 서의 활용가능성을 평가하였다. 상백피의 에탄올 추출물 (MRE)은 α-amylase와 α-glucosidase, pancreatic lipase를 활용한 소화효소 저해활성 평가 실험에서 각각 7.
본 실험결과 상백피 추출물은 전지방세포의 분화와 세포내 지질합성을 저해하고 비만과 관련된 소화효소에 대한 저해활성을 나타내었다. 이러한 결과를 기반으로 상백피의 비만 예방 소재로서의 잠재적인 가능성을 확인하였다.
가설 설정
2)ND : Not detected.
제안 방법
Cells were then maintained in DMEM supplemented with 10% FBS and 5μg/mL insulin for another 2 days (day 4), followed by culturing with DMEM with 10% FBS for an additional 4 days (day 8).
3T3-L1 preadipocyte cells have been served as a well-established in vitro model to search for new health benefit food or agents for the obesity and related metabolidisorders in numerous studies (34). In this study, first of all, the effects of mulberry root bark extracts (MRE and MRW) on cell viability and the differentiation of 3T3-L1 cells were evaluated by MTT assay and Oil Red O staining assay. As evidenced by the MTT assay, no significant cytotoxicity was observed at concentrations up to 400 μg/mL, as compared with the non-treated control (Fig.
대상 데이터
3T3-L1 cells were purchased from American Type Culture Collection (ATCC). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), calf bovine serum (Calf), phosphate-buffered saline (PBS, pH 7.
데이터처리
All values were expressed in terms of mean±standard deviation (SD) of at least three independent experiments, tested by analysis of variance (ANOVA) with Duncan’s multiple range tests.
Data are expressed in terms of mean±standard deviation (SD) of at least three independent experiments, tested by analysis of variance (ANOVA) with Duncan’s multiple range tests.
The percent inhibition of digestive enzymes was calculated as follows: Inhibition (%)=[(ODcontrol-ODsample)/ODcontrol]×100. The IC50 values were calculate by linear regression analysis.
이론/모형
(A) Cell viability of 3T3-L1 cells exposed to increasing concentration of MRW and MRE. Cells were treated with different concentration of MRW and MRE for 48 h and cell viability was analyzed using an MTT assay. (B) 3T3-L1 preadipocyte were induced to differentiate into adipocytes by the MDI [0.
The cytotoxicity of the sample was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method (29). Briefly, the cells were seeded at 104cells/well onto flat bottomed 96-well culture plates and treated with test samples at various concentrations.
성능/효과
The extracts were then freeze-dried and stored at -70℃ until used. The extraction yield obtained from MRE and MRW were 16.71% and 17.23% (w/w), respectively.
상백피의 항비만 작용 기전을 구명하기 위하여 adipogenesis 및 lipogenesis와 관련된 유전자 발현양상을 분석한 결과, 상백피 추출물 처리군에서는 생체내 지방대사 조절에 중요한 역할을 하는 FAS와 ACC 뿐 아니라 adipogenesis와 lipogenesis와 관련된 주요 전사요소인 PPARγ와 C/EBPα, SREBP-1c의 유전자 발현이 현저하게 억제되었다. qRT-PCR 분석 결과, 상백피 추출물의 anti-adipogenesis 효능은 전사단계에서의 관련 유전자 발현억제에 기인한다고 판단되었다. 본 실험결과 상백피 추출물은 전지방세포의 분화와 세포내 지질합성을 저해하고 비만과 관련된 소화효소에 대한 저해활성을 나타내었다.
qRT-PCR 분석 결과, 상백피 추출물의 anti-adipogenesis 효능은 전사단계에서의 관련 유전자 발현억제에 기인한다고 판단되었다. 본 실험결과 상백피 추출물은 전지방세포의 분화와 세포내 지질합성을 저해하고 비만과 관련된 소화효소에 대한 저해활성을 나타내었다. 이러한 결과를 기반으로 상백피의 비만 예방 소재로서의 잠재적인 가능성을 확인하였다.
상백피의 에탄올 추출물 (MRE)은 α-amylase와 α-glucosidase, pancreatic lipase를 활용한 소화효소 저해활성 평가 실험에서 각각 7.86±0.36, 0.12±0.03, 7.93±0.1 mg/mL 의 IC50 값을 보이며 우수한 억제 활성을 나타냈다.
상백피의 항비만 작용 기전을 구명하기 위하여 adipogenesis 및 lipogenesis와 관련된 유전자 발현양상을 분석한 결과, 상백피 추출물 처리군에서는 생체내 지방대사 조절에 중요한 역할을 하는 FAS와 ACC 뿐 아니라 adipogenesis와 lipogenesis와 관련된 주요 전사요소인 PPARγ와 C/EBPα, SREBP-1c의 유전자 발현이 현저하게 억제되었다.
후속연구
However, the deep investigation of molecular mechanism of MRE on PPARγ upstream signaling pathway need to be studied further.
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