[논문]죽력의 3T3-L1 지방전구세포에서 지방축적 억제 효과

장병철

doi:10.15429/jkomor.2022.22.1.38

죽력의 3T3-L1 지방전구세포에서 지방축적 억제 효과
Suppression of Lipid Accumulation in Differentiating 3T3-L1 Preadipocytes by a Standardized Commercial Juknyeok 원문보기

한방비만학회지 = Journal of Korean Medicine for Obesity Research, v.22 no.1, 2022년, pp.38 - 46

장병철 (계명대학교 의과대학 분자의학교실)

Abstract ▼ AI-Helper

Objectives: Juknyeok (JN) is natural liquor extracted from bamboo stems (Phyllostachys bambusoides) and has been used as a traditional Korean medicine for improving vascular function, blood glucose, and treating stroke. Until now, the JN's lipid-lowering effect and underlying mechanism in adipocytes are poorly understood. The aim of this study was to scrutinize the effect of a standardized commercial JN on lipid accumulation during the differentiation of 3T3-L1 preadipocytes. Methods: Lipid and triglyceride (TG) accumulation in differentiating 3T3-L1 preadipocytes were measured by Oil Red O staining and AdipoRed assay, respectively. Cell count analysis was used to ascertain 3T3-L1 cytotoxicity. Immunoblotting and Reverse transcription polymerase chain reaction analysis were used to assess protein and messenger RNA (mRNA) expression levels in 3T3-L1 cells, respectively. Results: Treatment with JN at 25 𝜇l/ml after pH calibration with 6.35 significantly reduced lipid and TG accumulation in differentiating 3T3-L1 preadipocytes without significant cytotoxicity. On mechanistic levels, JN markedly suppressed protein expression levels of CCAAT/enhancer-binding protein (C/EBP)-𝛽 and fatty acid synthase (FAS) during the differentiation of 3T3-L1 preadipocytes. However, JN did not affect the protein expression levels of C/EBP-𝛼, peroxisome proliferator-activated receptor-𝛽/𝛾, and phosphorylation levels of signal transducer and activator of transcription-3/5 in differentiating 3T3-L1 preadipocytes. JN also reduced leptin mRNA expression levels in differentiating 3T3-L1 preadipocytes. Conclusions: JN at 25 𝜇l/ml lowers lipid accumulation and TG content in differentiating 3T3-L1 cells, mediated through the reduced expression levels of C/EBP-𝛽 and FAS.

주제어

표/그림 (4)

그림 Fig. 1. Effects of JN at different concentrations on intracellular lipid accumulation in differentiating 3T3-L1 cells. (A) The experimental scheme of 3T3-L1 preadipocytes differentiation. (B) 3T3-L1 preadipocytes were induced to differentiate with an induction medium containing MDI, insulin, and FBS in the presence or absence of JN at the indicated concentrations for 8 days. On D8, the intracellular LDs were assessed by Oil Red O staining (upper panels in [B]). Phase-contrast images of the control or JN-treated cells on D8 were also taken (lower panels in [B]). JN: Juknyeok, MDI: 0.5 mM IBMX, 0.5 μM dexamethasone, and 5 μg/mL insulin, FBS: fetal bovine serum, D: day, LD: lipid droplets.
그림 Fig. 2. Effects of JN at different concentrations on intracellular TG content and cell growth in differentiating 3T3-L1 cells. (A, B) 3T3-L1 preadipocytes were induced to differentiate with an induction medium containing MDI, insulin, and FBS in the presence or absence of JN at the indicated concentrations for 8 days. On D8, intracellular TG content was measured by AdipoRed assay (A). Values are mean±standard error of data from three independent experiments with triplicate. *P<0.05 vs control. On D8, the control or JN-treated 3T3-L1 cells, which cannot be stained with trypan blue dye, were counted under a microscope (B). The cell count assay was done in triplicates. Data are mean±SE of three independent experiments. *P<0.05 vs control. JN: Juknyeok, TG: triglyceride, MDI: 0.5 mM IBMX, 0.5 μM dexamethasone, and 5 μg/mL insulin, FBS: fetal bovine serum, D: day, LD: lipid droplets.
그림 Fig. 3. Effects of JN at 25 μl/ml on expression and phosphorylation levels of C/EBP-α, C/EBP-β, PPAR-γ, PPAR-β, STAT-3, and STAT-5 in differentiating 3T3-L1 cells. 3T3-L1 preadipocytes were induced to differentiate with an induction medium containing MDI, insulin, and FBS in the presence or absence of JN at 25 μl/ml, and harvested at D2, D5, and D8, respectively. Whole-cell lysates at the indicated time point were extracted and analysed by Western blot analysis with respective antibodies. p-STAT-3, phosphorylated STAT-3; T-STAT-3, total STAT-3; p-STAT-5, phosphorylated STAT-5; T-STAT-5, total STAT-5. JN: Juknyeok, C/EBP: CCAAT/enhancerbinding proteins, PPAR: peroxisome proliferator-activated receptors, STAT: signal transducer and activator of transcription, MDI: 0.5 mM IBMX, 0.5 μM dexamethasone, and 5 μg/mL insulin, FBS: fetal bovine serum, D: day.
그림 Fig. 4. Effects of JN at 25 μl/ml on protein and mRNA expression levels of FAS and leptin in differentiating 3T3-L1 cells. (A, B) 3T3-L1 preadipocytes were induced to differentiate with an induction medium containing MDI, insulin, and FBS in the presence or absence of JN at 25 μl/ml, and harvested at D2, D5, and D8, respectively. Whole-cell lysates and total cellular mRNA at the indicated time point were extracted and analysed by Western blot (A) and RT-PCR (B) analysis, respectively. JN: Juknyeok, mRNA: messenger RNA, FAS: fatty acid synthase, MDI: 0.5 mM IBMX, 0.5 μM dexamethasone, and 5 μg/mL insulin, FBS: fetal bovine serum, D: day, RT-PCR: reverse transcription polymerase chain reaction.

참고문헌 (33)

Fruh SM. Obesity: risk factors, complications, and strategies for sustainable long-term weight. J Am Assoc Nurse Pract. 2017 ; 29(1) : S3-S14.
Aranaz P, Herrera DN, Zabala M, Migueliz I, Hualde AR, Yoldi ML, et al. Phenolic compounds inhibit 3T3-L1 adipogenesis depending on the stage of differentiation and their binding affinity to PPARγ. Molecules. 2019 ; 24(6) : 1045.
Ellulu MS, Patimah I, Khazaai H, Rahmat A, Abed Y. Obesity and inflammation: the linking mechanism and the complications. Arch Med Sci. 2017 ; 13(4) : 851-63.

상세보기
Ma X, Lee P, Chisholm DJ, James DE. Control of adipocyte differentiation in different fat depots; implications for pathophysiology or therapy. Front Endocrinol. 2015 ; 6(1) : 1-8.
Berndt J, Kovacs P, Ruschke K, Kloting N, Fasshauer M, Schon MR, et al. Fatty acid synthase gene expression in human adipose tissue: association with obesity and type 2 diabetes. Diabetologia. 2007 ; 50 : 1472-80.

상세보기
Cordonier EL, Jarecke SK, Hollinger FE, Zempleni J. Inhibition of acetyl-CoA carboxylases by soraphen A prevents lipid accumulation and adipocyte differentiation in 3T3-L1 cells. Eur J Pharmacol. 2016 ; 780 : 202-8.

상세보기
Kern PA, Di Gregorio G, Lu T, Rassouli N, Ranganathan G. Perilipin expression in human adipose tissue is elevated with obesity. J Clin Endocrinol Metab. 2004 ; 89 : 1352-8.

상세보기
Panee J. Potential medicinal application and toxicity evaluation of extracts from bamboo plants. J Med Plant Res. 2015 ; 9 : 681-92.
Park EJ, Jhon DY. Effects of bamboo shoot consumption on lipid profiles and bowel function in healthy young women. Nutrition. 2009 ; 25(7-8) : 723-8.
Lee MJ, Park WH, Song YS, Lee YW, Song YO, Moon GS. Effect of bamboo culm extract on oxidative stress and genetic expression: bamboo culm extract ameliorates cell adhesion molecule expression and NFkappaB activity through the suppression of the oxidative stress. Clin Nutr. 2008 ; 27 : 755-63.
Choi MH, Jo HG, Yang JH, Ki SH, Shin HJ. Antioxidative and anti-melanogenic activities of bamboo stems (Phyllostachys nigra variety henosis) via PKA/CREB-mediated MITF downregulation in B16F10 melanoma cells. Int J Mol Sci. 2018 ; 19 : 409.
Jakab J, Miskic B, Miksic S, Juranic B, Cosic V, Schwarz D, et al. Adipogenesis as a potential anti-obesity target: a review of pharmacological treatment and natural products. Diabetes Metab Syndr Obes. 2021 ; 14 : 67-83.

상세보기
Jayarathne S, Koboziev I, Park OH, Oldewage-Theron W, Shen CL, Moustaid-Moussa N. Anti-inflammatory and anti-obesity properties of food bioactive components: effects on adipose tissue. Prev Nutr Food Sci. 2017 ; 22(4) : 251-62.

상세보기
Cao Z, Umek RM, McKnight SL. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 1991 ; 5(9) : 1538-52.

상세보기
Farmer SR. Transcriptional control of adipocyte formation. Cell Metab. 2006 ; 4(4) : 263-73.
Rosen ED, Sarraf P, Troy AE, Bradwin G, Moore K, Milstone DS, et al. PPARγ is required for the differentiation of adipose tissue in vivo and in vitro. Mol Cell. 1999 ; 4(4) : 611-7.
Linhart HG, Ishimura-Oka K, DeMayo F, Kibe T, Repka D, Poindexter B, et al. C/EBPα is required for differentiation of white, but not brown, adipose tissue. Proc Natl Acad Sci USA. 2001 ; 98(22) : 12532-7.
Stephens JM, Morrison RF, Pilch PF. The expression and regulation of STATs during 3T3-L1 adipocyte differentiation. J Biol Chem. 1996 ; 271(18) : 10441-4.

상세보기
Rosen ED, Hsu CH, Wang X, Sakai S, Freeman MW, Gonzalez FJ, et al. C/EBPα induces adipogenesis through PPARγ: a unified pathway. Genes Dev. 2002 ; 16(1) : 22-6.
Deng J, Hua K, Lesser SS, Harp JB. Activation of signal transducer and activator of transcription-3 during proliferative phases of 3T3-L1 adipogenesis. Endocrinology. 2000 ; 141 : 2370-6.

상세보기
Harp JB, Franklin D, Vanderpuije AA, Gimble JM. Differential expression of signal transducers and activators of transcription during human adipogenesis. Biochem Biophys Res Commun. 2001 ; 281 : 907-12.

상세보기
Stewart WC, Morrison RF, Young SL, Stephens JM. Regulation of signal transducers and activators of transcription (STATs) by effectors of adipogenesis: coordinate regulation of STATs 1, 5A, and 5B with peroxisome proliferator-activated receptor-γ and C/AAAT enhancer binding protein-α. Biochim Biophys Acta. 1999 ; 1452 : 188-96.
Shang CA, Waters MJ. Constitutively active signal transducer and activator of transcription 5 can replace the requirement for growth hormone in adipogenesis of 3T3-F442A preadipocytes. Mol Endocrinol. 2003 ; 17 : 2494-508.

상세보기
Ho CL, Lin CY, Ka SM, Chen A, Tasi YL, Liu ML, et al. Bamboo vinegar decreases inflammatory mediator expression and NLRP3 inflammasome activation by inhibiting reactive ocygen species generation and protein kinase C-α/δ activation. Plos One. 2013 ; 8(10) : e75738.
Higa JK, Panee J. Bamboo extract reduces interleukin 6 (IL-6) overproduction under lipotoxic conditions through inhibiting the activation of NF-kB and AP-1 pathways. Cytokine. 2011 ; 55 : 18-23.

상세보기
Greenberg AS, Obin MS. Obesity and the role of adipose tissue in inflammation and metabolism. The American Journal of Clinical Nutrition. 2006 ; 83 (2) : 461S-5S.
Maffei M, Halaas J, Ravussin E, Pratley RE, Lee GH, Zhang Y, et al. Leptin levels in human and rodent: measurement of plasma leptin and ob RNA in obese and weight-reduce subject. Nat Med. 1995 ; 1 : 1155-61.
Halaas JL, Gajiwala KS, Maffei M, Cohen SL, Chait BT, Rabinowitz D, et al. Weight-reducing effects of the plasma protein encoded by the obese gene. Science. 1995 ; 269 : 543-6.

상세보기
Pelleymounter MA, Cullen MJ, Baker MB, Hecht R, Winters D, Boone T, et al. Effects of the obese gene product on body weight regulation in ob/ob mice. Science. 1995 ; 269 : 540-3.

상세보기
Akakabe Y, Tamura Y, Iwamoto S, Takabayashi M, Nyuugaku T. Volatile organic compounds with characteristic odo in bambbo vinegar. Biosci Biotechnol Biochem. 2006 ; 70(11) : 2797-9.
Mu J, Uehara T, Furno T. Effect of bamboo vinegar on regulation of germination and radicle growth of seed plants. J Wood Sci. 2003 ; 49 : 262-70.
Heimann E, Nyman M, Degerman E. Propionic acid and butyric aicid inhibit lipolysis and de novo lipogenesis and increase insulin-stimulated glucose uptake in primary rat adipocytes. Adipocyte. 2015 ; 4(2) : 81-8.
Farup PG, Lydersen S, Valeur J. Are nonnutritive sweeteners obesogenic? Associatios between diet, faecal microbiota, and short-chain fatty acids in morbidly obese subjects. Journal of Obesity. 2019 ; 2019 : 4608315.

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증