[논문]Atorvastatin으로 유도된 근위축 세포모델에서 상황-오미자박 고상발효물 열수추출물의 보호효과

김영숙; 황수진; 박광일; 임종민; 천다미; 정유진; 전병엽; 곽경태; 오태우

doi:10.14374/hfs.2021.29.4.239

Atorvastatin으로 유도된 근위축 세포모델에서 상황-오미자박 고상발효물 열수추출물의 보호효과
Protective Effect of water extract Phellinus linteus-discard Schisandra chinensis solid fermented extracts on improvement of sarcopenia by Atorvastatin-induced muscle atrophy cell model 원문보기

大韓韓醫學方劑學會誌 = Herbal formula science, v.29 no.4, 2021년, pp.239 - 252

김영숙 ((주)글루칸) , 황수진 (한국한의학연구원 한의기술응용센터) , 박광일 (경상국립대학교 수의과대학) , 임종민 ((주)글루칸) , 천다미 ((주)글루칸) , 정유진 (한국화학연구원 정밀화학융합기술연구센터) , 전병엽 ((주)큐비엠) , 곽경태 ((주)큐비엠) , 오태우 (한국한의학연구원 한의기술응용센터)

Abstract ▼ AI-Helper

Objectives : This study is to effect of improving muscle atrophy through water extract on the solid-phase fermentation extraction with Phellinus linteus of discarded Schisandra chinensis in an atorvastatin-induced atrophy C2C12 cell. Methods : C2C12 myoblast were differentiated into myotube by 2% horse serum medium for 6 days, and then treated solid-phase fermentation(S-P) extract at different concentrations for 24h. To investigate the effect of S-P extract on the induction of muscle atrophy and expression of atrophy-related genes and apoptosis in differentiated C2C12 myotubes using a GSH, ROS, real-time PCR, western blots analysis. Results : As a result of treatment with atorvastatin at concentrations of 5, 10, and 20 uM on the 6th day of differentiation in C2C12 myotube cells, it was confirmed that the cell morphology was damaged in a concentration-dependent manner, and the length and thickness of the myotube also decreased in a concentration-dependent manner. Treatment with S-P extract (50, 100 and 200 ㎍/㎖) increased of GSH and inhibited ROS in the atorvastatin-induced muscle atrophy cell model at a concentration that did not induce toxicity. In addition, it was confirmed that it has an effect on muscle reduction by inhibiting apoptosis of muscle cells as well as being involved in protein production and degradation of muscle cells. Conclusions : Atorvastatin-induced atrophy C2C12 cell, S-P extract activates related to differentiation/generation and proteolysis, and inhibits cell death of atrophy in C2C12 cell. Based on this, it is necessary to prove its effectiveness through animal models and human application test, but it is considered to be discarded Schisandra chinensis can present the potential for development as a recycling industrial material.

주제어

표/그림 (7)

표 Table 1. Specific primer sequences for PCR
그림 Fig. 1. Differentiation of C2C12 myoblasts into myotubes. (A) Differentiation of myoblasts was induced for 8 days in differentiation medium containing 2% HS, and differentiation medium was changed every 2 days (B) Myotube lengh and widths in C2C12 myotubes. Values are means SEM of two different preparations with quadruplicate experiments. ^*p<0.05, ^**p<0.01 and ^***p<0.001 vs. CTL group.
그림 Fig. 2. Atorvastatin(ATV) treatment after differentiation induced myotube atrophy. (A) Change of cell morphology C2C12 myotubes treated with 0, 5, 10, and 20 uM ATV after differentiation. (B) Measurement of Myotube lengh and widths treated ATV in C2C12 myotubes. (C) Changes of cell morphology treated with ATV for 24 h and 48 h in C2C12 cell. (D) Measurement of Myotube lengh and widths treated ATV for 24 h and 48 h in C2C12 cell. Values are means SEM of two different preparations with quadruplicate experiments. ^**p<0.05, ^**p<0.01 and ^***p<0.001 vs. CTL group.
그림 Fig. 3. Cell viability of C2C12 cells treated S-P extract. (A) Cytotoxic effects of S-P extract in C2C12 cell. (B) Change of cell morphology in C2C12 myotubes treated with 10 uM ATV and 200 ㎍/㎖ S-P extract after differentiation. (C) Measurement of Myotube lengh and widths treated ATV for 24 h and 48 h in C2C12 cell. C2C12 cells were treated with ATV (10 uM) with S-P extract (200 ㎍/㎖) for 24h. Values are means SEM of two different preparations with quadruplicate experiments. ^###p<0.001 vs. CTL ; ^**p<0.01 vs. ATV group.
그림 Fig. 4. The Effect of S-P extract on GSH(A) and ROS(B) levels in C2C12 myotubes. C2C12 cells were treated with ATV (10 uM) with various concentration of S-P extracts (50, 100 and 200 ㎍/㎖) for 24h. Values are means SEM of two different preparations with quadruplicate experiments. ^###p<0.001 vs. CTL ; ^**p<0.01 and ^***p<0.001 vs. ATV group.
그림 Fig. 5. Effect of S-P extract on the Atorvastatin induced muscle atrophy in C2C12 myotube. Total RNAs were extracted using Trizol reagent and mRNA was measured using qRT-PCR. The fold changes relative to the control were calculated using the ΔΔCT method for mRNA expression levels of muscle-specific transcription genes. Values are means SEM of two different preparations with quadruplicate experiments. ^**p<0.01 and ^***p<0.001 vs. CTL ; ^#p<0.05, ^##p<0.01 and ^###p<0.001 vs. ATV group.
그림 Fig. 6. Atrophy-related apoptosis in ATV treated C2C12 myotubes. The expression of Bax/Bcl2 and cleaved-caspase-3/caspase-3 mRNA and proteins expression by qRT-PCR and western blot. Cells treated with S-P extract (50, 100 and 200 ㎍/㎖) in the absence or presence of 10uM ATV for 24 h. The fold changes relative to the control were mRNA expression levels of apoptosis genes and the intensity of the protein band was normalized against the internal control β-actin. Values are means SEM of two different preparations with quadruplicate experiments. ^***p<0.001 vs. CTL ; ^##p<0.01 and ^###p<0.001 vs. ATV group.

참고문헌 (30)

Fried LP, Xue QL, Cappola AR, Ferrucci L, Chaves P, Varadhan R, et al. Nonlinear multisystem physiological dysregulation associated with frailty in older women: implications for etiology and treatment. J Gerontol A Biol Sci Med Sci. 2009; 64:1049-1057.
Wilson D, Jackson T, Sapey E, Lord JM. Frailty and sarcopenia: The potential role of an aged immune system. Ageing Res Rev 2017;36:1-10.

상세보기
Fried LP, Tangen CM, Walston J, Newman AB, Hirsch C, Gottdiener J, et al. Frailty in older adults: Evidence for a phenotype. J Gerontol: Series A. 2001;56:M146-M157.
Habukuihakwon. youngchu-kyungkyosuk. Beijing: Inminwisheng publisher; 1982:161.24.
SJ Hwang, JM Lim, BH Ku, DM Cheon, YJ Jung, YS Kim, TW Oh. Effects of polysaccharide (polycan) derived from black yeast in dexamethasone-induced muscle atrophy cell model. Herb Formula Sci. 2021;29(1):45-55.
Shepherd J, Blauw GJ, Murphy MB, Bollen EL, Buckley BM, Cobbe SM, et al. Pravastatin in elderly individuals at risk of vascular disease (PROSPER): a randomised controlled trial. Lancet 2002;360:1623-30.
Shelina MJ, Mark JE, Stavroula C. Rhabdomyolysis associated with hyeroxymethylglutaryl-coenzyme A reductase inhibitors. Am Heart J. 2004;147: 956-965.
Stamler J, Wnetworth D, Neaton JD. Is relationship between serum cholesterol and risk of premature death from coronary heart disease continuous and graded? Findings in 356,222 primary screens of the Multiple Risk Factor Intervention Trial (MRFIT). JAMA. 1986;256:2823-2828.
Heart Protection Study Collaborative Group. MRC/BHF Heart Protection Study of cholesterol-lowering with simvastatin in 5963 people with diabetes: a randomised placebo-controlled trial. Lancet. 2003; 361:2005-2016.
Huang F, Xiong Y, Xu L, Ma S, Dou C. Sedative and hypnotic activities of the ethanol fraction from Fructus Schisandrae in mice and rats. J Ethnopharmacol. 2007:4;110(3):471-475.

상세보기
Wei B, Li Q, Su D, Fan R, Zhao L, Geng L, He B, Chen X, Jia Y, Bi K. Development of a UFLC-MS/MS method for simultaneous determination of six lignans of Schisandra chinensis (Turcz.) Baill. in rat plasma and its application to a comparative pharmacokinetic study in normal and insomnic rats. J Pharm Biomed Anal. 2013; 15:77:120-127.
Panossian A, Wikman G. Pharmacology of Schisandra chinensis Bail.: an overview of Russian research and uses in medicine. J Ethnopharmacol. 2008; 23;118(2);183-212.

상세보기
Lee JY, Min YK, Kim HY. Isolation of antimicrobial substance from Schizandra chinensis Baillon and antimicrobial effect. Korean J Food Sci Technol. 2001;33:389-394.
Yeo SG, Ahn CW, Lee YW, Lee TG, Park YH, Kim SB. Antioxidative effect of tea extracts from green tea, oolong tea and black tea. J Korean Soc Food Nutr. 1995l24:299-304.
Ko BS, Park SK, Choi SB, Jun DW, Choi MK, Park SM. A study on hypoglycemic effects of crude extracts of Schizandrae Fructus. J Korean Soc Appl Biol Chem. 2004;47:258-264.
Chung KH, Lee SH, Lee YC, Kim JT. Antimicrobial activity of Omija (Schizandra chinensis) extracts. J Korean Soc Food Sci Nutr. 2001;30:127-132.
Zhu M, Lin KF, Yeung RY, Li RC. Evaluation of the protective effects of Schisandra chinensis on phase I drug metabolism using a CCl4 intoxication model. J Ethnopharmacol. 1999;67:61-68.

상세보기
Xing J, Guo Y, Hu H, Qu XL, Sun XZ, Liu SH, Wang H. A herbal composition of Semen Hoveniae, Radix Puerariae, and Fructus Schisandrae shows potent protective effects on acute alcoholic intoxication in rodent models. Evid Based Complement Alternat Med. 2012;638197. doi: 10.1155/2012/638197.

상세보기
Cheng N, Ren N, Gao H, Lei X, Zheng J, Cao W. Antioxident and hepatoprotective effects of Schisandra chinensis pollen extract on CCl4 induced acute liver damage in mice. Food Chem Toxicol. 2013;55:234-240.
Lu Y, Chen DF. Analysis of Schisandra chinensis and Schisandra sphenanthera. J Chromatogr A. 2009;13(11):1980-1990.
Cebron Lipovec N, Schols AMWJ, van den Borst B, Beijers RJHCG, Kosten T, Omersa D, Lainscak M. Sarcopenia in advanced COPD affects cardiometabolic risk reduction by shortterm high-intensity pulmonary rehabilitation. J Am Med Dir Assoc. 2016;17(9):814-820.

상세보기
Park SH, Park JH, Song PS, Kim DK, Kim KH, Seol SH, Kim HK, Jang HJ, Lee JG, Park HY, Park JS, Shin KJ, Kim DI, Moon YS. Sarcopenic obesity as an independent risk factor of hypertension. J Am Soc Hypertens. 2013;7(6): 420-425.
Esther A, Joan AB, Josep R, Jose CF. Glutathione Depletion Impairs Myogenic Differentiation of Murine Skeletal Muscle C2C12 Cells through Sustained NF-κB Activation. Am J Pathol. 2004;165(3):719-728.
Bentzinger CF, Wang YX, Rudnicki MA. Building muscle: molecular regulation of myogenesis. Cold Spring Harb Perspect Biol. 2012;4(2). pii: a008342.
Gomes MD, Lecker SH, Jagoe RT, Navon A, Goldberg AL. Atrogin-1, a muscle-specific F-box protein highly expressed during muscle atrophy. Proc Natl Acad Sci USA 2001;98:14440-14445.

상세보기
Bodine SC, Latres E, Baumhueter S, et al. Identification of ubiquitin ligases required for skeletal muscle atrophy. Science 2001;294:1704-1708.

상세보기
Williamson DL, Raue U, Slivka DR, Trappe S. Resistance exercise, skeletal muscle FOXO3A, and 85-year-old women. J Gerontol A Biol Sci Med Sci 2010;65:335-343.
Braga M, Sinha Hikim AP, Datta S, et al. Involvement of oxidative stress and caspase 2-mediated intrinsic pathway signaling in age-related increase in muscle cell apoptosis in mice. Apoptosis 2008;13:822-832.

상세보기
Dupont-Versteegden EE. Apoptosis in muscle atrophy:relevance to sarcopenia. Exp Gerontol 2005;40:473-481.

상세보기
Yeh TS, Hsu CC, Yang SC, Hsu MC, Liu JF. Angelica Sinensis promotes myotube hypertrophy through the PI3K/Akt/mTOR pathway. BMC Complement Altern Med. 2014;14(1):144.

저자의 다른 논문 :

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].

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