[논문]Effect of palmitoleic acid on the differentiation of bovine skeletal muscle satellite cells

Zhang, Junfang; Li, Qiang; Nogoy, Kim Margarette Corpuz; Sun, Jianfu; Sun, Bin; Wang, Ying; Tang, Lin; Yu, Jia; Jin, Xin; Li, Xiangzi; Choi, Seong-Ho

doi:10.5187/jast.2021.e78

Effect of palmitoleic acid on the differentiation of bovine skeletal muscle satellite cells 원문보기

Journal of animal science and technology : JAST, v.63 no.4, 2021년, pp.919 - 933

Zhang, Junfang (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Li, Qiang (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Nogoy, Kim Margarette Corpuz (Department of Animal Science, Chungbuk National University) , Sun, Jianfu (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Sun, Bin (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Wang, Ying (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Tang, Lin (Engineering Research Center of North-East Cold Region Beef Cattle Science & Technology Innovation, Ministry of Education, Yanbian University) , Yu, Jia (Department of Animal Science, Chungbuk National University) , Jin, Xin (Engineering Research) , Li, Xiangzi , Choi, Seong-Ho

Abstract ▼ AI-Helper

We hypothesized that the unsaturated fatty acid palmitoleic acid (POA) could promote the expression of adipogenic/lipogenic genes in bovine skeletal muscle satellite cells (BSCs). The BSCs were cultured in a growth medium containing 10% fetal bovine serum. When the cells reached 80%-90% confluence, we used the differentiation medium with 5% horse serum for differentiation for 96 h. The differentiation medium contained 50 µM, 100 µM and 200 µM POA. Control BSC were cultured only in differentiation media. Compared with the control BSC, the POA BSC significantly up-regulated the expression of paired box 3 (Pax3) and paired box 7 (Pax7) and down-regulated myogenin gene expression (p < 0.01), which indicates a depression in muscle fiber development. However, all POA treatments up-regulated the expression of the adipocyte transcription factors peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein alpha and beta (C/EBP α and C/EBP β), and other genes (p < 0.01) and increased the expression of PAT-family proteins and the concentration of adiponectin in the media. These results indicate that POA can convert part of BSCs into adipocytes.

주제어

표/그림 (7)

표 Table 1. Forward and reverse primers for real-time PCR for specifc gene mRNA
그림 Fig. 1. Effects of different concentrations of palmitoleic acid (POA) on average cell size (A) and cell viability (B) in bovine satellite cells after 96-h culture. Control group (CON), DMEM + 5% horse serum; POAL, CON + 50 μM POA; POAM, CON + 100 μM POA; POAH, CON + 200 μM POA. Values with a different letter (a or b or c) above column differ significantly (p 0.05). DMEM, Dulbecco's Modified Eagle's medium.
그림 Fig. 2. Identification of Yanbian Yellow Cattle BSC. (A) (a) Cells began to adhere to the wall and grew in a long shuttle shape; (b) Cell fusion occurs and multinuclear myotubes begin to form. The arrows indicate the fused cells. (B) The expressions of Pax7, Desmin and MyoD were identified by immunofluorescence staining. After DAPI staining, the nuclei were blue, and the cytoplasm of Pax7, Desmin and MYOD were green. The magnification is ×40 (scale bars = 200 μm). DAPI, 4',6-diamidino-2-phenylindole; Pax7, paired box 7; MyoD, myogenic differentiation antigen; BSC, bovine satellite cells.
그림 Fig. 3. Oil-red-o, lipid droplet area, intracellular triacylglycerol, and lipid droplet area analyses after differentiation of BSC for 96h. (A) Oil red staining after 96-h incubation of BSC treated with POA (scale bars = 200 μm), (B) effects of different concentrations of POA on lipid droplet area in BSC at 96-h culture tim, (C) Effects of different concentrations of POA on intracellular triacylglycerol in BSC at 96-h culture time, (D) adiponectin concentration (μg/L) in the medium after 96 h differentiation culture of BSC with POA. CON, DMEM + 5% HS; POAL, CON + 50 μM POA; POAM, CON + 100 μM POA; POAH, CON + 200 μM POA. BSC, bovine satellite cells; POA, palmitoleic acid; DMEM, Dulbecco's Modified Eagle's medium; HS, horse serum.
그림 Fig. 4. Expression of myogenic genes and proteins. (A) Pax3, (B) Pax7, (C) MYOD, (D) MYOG, (E) MYF5, (F) MRF4, (G) expression of the indicated proteins was analyzed by western blot after treatment of the BSC with POA for 96 h. CON, DMEM + 5% HS; POAL, CON + 50 μM POA; POAM, CON + 100 μM POA; POAH, CON + 200 μM POA. Pax3, paired box 3; Pax7, paired box 7; MYOD, myogenic differentiation antigen; MYOG, myogenin; MYF5, myofactor-5; MRF4, myofactor-4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; BSC, bovine satellite cells; POA, palmitoleic acid; DMEM, Dulbecco's Modified Eagle's medium; HS, horse serum.
그림 Fig. 5. Expression of adipogenic genes and proteins. (A) PPARγ, (B) C/EBPα, (C) C/EBPβ, (D) SREBP1, (E) GPR43, (F) CPT1β), (G) expression of the indicated proteins was analyzed by western blot after treatment of the BSC with POA for 96 h. CON, DMEM + 5% HS; POAL, CON + 50 μM POA; POAM, CON + 100 μM POA; POAH, CON + 200 μM POA. PARγ, peroxisome proliferator-activated receptor gamma; C/EBPα, CCAAT/enhancer-binding protein alpha; SREBP1, sterol-regulated binding protein 1; C/EBPβ, CCAAT/enhancer-binding protein beta; GPR43, G-protein coupled receptor 43; CPT1β, carnitine palmitoyltransferase 1 beta; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; BSC, bovine satellite cells; POA, palmitoleic acid; DMEM, Dulbecco's Modified Eagle's medium; HS, horse serum.
그림 Fig. 6. Expression of lipid metabolism-associated genes. (A) SCD, (B) AMPKα, (C) LPL, (D) FASN, (E) GPAT3, (F) ACSL3, (G) PLIN1, (H) PLIN2, (I) PLIN3, (J) CD36, (K) Leptin, (L) ADI. CON, DMEM + 5% HS; POAL, CON + 50 μM POA; POAM, CON + 100 μM POA; POAH, CON + 200 μM POA. SCD, stearoyl coenzyme-A desaturation enzyme; AMPKα, AMP-activated protein kinase alpha-1; LPL, lipoprotein lipase; FASN, fatty acid synthase; GPAT3, glycerin-3-phosphate acyltransferase 3; ACSL3, acyl-CoA synthetase long-chain 3; PLIN1, lipid droplet formation-related protein, perilipin; PLIN2, adipose differentiation-related protein (adipophilin); PLIN3, tail-interacting protein of 47 kDa (TIP47); ADI, adiponectin; DMEM, Dulbecco's Modified Eagle's medium; HS, horse serum; POA, palmitoleic acid.

AI 본문요약
AI-Helper

데이터처리

Significant differences among the treatments were analyzed using one-way analysis of variance and Post Hoc Tukey’s multiple comparison test GraphPad Prism 6.07 (GraphPad Software, La Jolla, CA, USA) at significance level of p < 0.05.

이론/모형

All experimental procedures involving animals were conducted using the approval code YBU- 20160303 provided by the Institutional Animal Care and Use Committee of Yanbian University.
All experimental procedures involving animals were conducted using the approval code YBU-20160303 provided by the Yanbian University Institutional Animal Care and Use Committee.

성능/효과

In conclusion, treatment of BSC with POA can promote the transdifferentiation of BSCs into adipocytes where at least a portion of BSCs were converted into adipocytes as evidenced by the downregulations of myogenic related genes that caused the depression of muscle fiber development, and upregulation of adipogenic related genes that caused the transdifferentiation of muscle cells into adipocyte-like cells. This study demonstrated that POA was could be a positive regulator for differentiation of BSCs to ADPs.
This study demonstrated that POA was could be a positive regulator for differentiation of BSCs to ADPs. Our results suggest that POA could be useful in increasing the marbling content of beef to improve meat quality and this also shows that bovine satellite cells could be useful study models in the development of meat culture technology.
Western blot analysis was used to determine the expression levels of PAX7, MYOD, and MYOG. Results showed that POA treatment increased the protein expression of myogenic transcription factor PAX7 and decreased the MYOD and MYOG in BSC (Fig. 4G).
The average cell size in CON + 100 μM POA ([POAM] 20.60 ± 0.50 μm) and CON + 200 μM POA ([POAH] 23.36 ± 0.39 μm) treatments was higher than that of CON (15.93 ± 0.27 μm) (p < 0.001), whereas there was no significant difference between CON (15.93 ± 0.27 μm) and CON + 50 μM POA ([POAL] 17.20 ± 0.20 μm) (Fig
In conclusion, treatment of BSC with POA can promote the transdifferentiation of BSCs into adipocytes where at least a portion of BSCs were converted into adipocytes as evidenced by the downregulations of myogenic related genes that caused the depression of muscle fiber development, and upregulation of adipogenic related genes that caused the transdifferentiation of muscle cells into adipocyte-like cells. This study demonstrated that POA was could be a positive regulator for differentiation of BSCs to ADPs. Our results suggest that POA could be useful in increasing the marbling content of beef to improve meat quality and this also shows that bovine satellite cells could be useful study models in the development of meat culture technology.

참고문헌 (50)

1. Ladeira MM Schoonmaker JP Swanson KC Duckett SK Gionbelli MP Rodrigues LM et al Review: nutrigenomics of marbling and fatty acid profile in ruminant meat Animal 2018 12 s282 94 10.1017/S1751731118001933 30139403

상세보기
2. Baik M Kang HJ Park SJ Na SW Piao M Kim SY et al Triennial Growth and Development Symposium: molecular mechanisms related to bovine intramuscular fat deposition in the longissimus muscle J Anim Sci. 2017 95 2284 303 10.2527/jas.2016.1160 28727015

상세보기
3. Chen J Liu M Li Z The fractal dimension research of chinese and american beef marbling standards images In Proceedings of the International Conference on Computer & Computing Technologies in Agriculture 2012 Oct 19-21 Zhangjiajie, China
4. Hocquette JF Gondret F Baeza E Medale F Jurie C Pethick DW Intramuscular fat content in meat-producing animals: development, genetic and nutritional control, and identification of putative markers Animal 2010 4 303 19 10.1017/S1751731109991091 22443885

상세보기
5. Mach N Bach A Realini CE Font IFM Velarde A Devant M Burdizzo pre-pubertal castration effects on performance, behaviour, carcass characteristics, and meat quality of Holstein bulls fed high-concentrate diets Meat Sci. 2009 81 329 34 10.1016/j.meatsci.2008.08.007 22064171

상세보기
6. Brooks M Choi C Lunt D Miller R Choi C Smith S Case study: carcass and meat characteristics and M. longissimus thoracis histology of beef from calf-fed and yearling-fed Angus steers Prof Anim Sci. 2011 27 385 93 10.15232/S1080-7446(15)30503-9

상세보기
7. Moody WG Cassens RG A quantitative and morphological study of bovine longissimus fat cells J Food Sci. 1968 33 47 52 10.1111/j.1365-2621.1968.tb00882.x

상세보기
8. Smith SB Lunt DK Marbling: management of cattle to maximize the deposition of intramuscular adipose tissue In Proceedings of the Plains Nutrition Council Spring Conference 2007 2007 Mar 29-30 San Antonio, Tx
9. Garmyn AJ Hilton GG Mateescu RG Morgan JB Reecy JM Tait RG Jr et al Estimation of relationships between mineral concentration and fatty acid composition of longissimus muscle and beef palatability traits J Anim Sci. 2011 89 2849 58 10.2527/jas.2010-3497 21512113

상세보기
10. Collins CA Olsen I Zammit PS Heslop L Petrie A Partridge TA et al Stem cell function, self-renewal, and behavioral heterogeneity of cells from the adult muscle satellite cell niche Cell 2005 122 289 301 10.1016/j.cell.2005.05.010 16051152

상세보기
11. Shefer G Wleklinski-Lee M Yablonka-Reuveni Z Skeletal muscle satellite cells can spontaneously enter an alternative mesenchymal pathway J Cell Sci. 2004 117 5393 404 10.1242/jcs.01419 15466890

상세보기
12. Csete M Walikonis J Slawny N Wei Y Korsnes S Doyle JC et al Oxygen-mediated regulation of skeletal muscle satellite cell proliferation and adipogenesis in culture J Cell Physiol. 2001 189 189 96 10.1002/jcp.10016 11598904

상세보기
13. Teboul L Gaillard D Staccini L Inadera H Amri EZ Grimaldi PA Thiazolidinediones and fatty acids convert myogenic cells into adipose-like cells J Biol Chem. 1995 270 28183 7 10.1074/jbc.270.47.28183 7499310

상세보기
14. Li XZ Yan Y Zhang JF Sun JF Sun B Yan CG et al Oleic acid in the absence of a PPAR γ agonist increases adipogenic gene expression in bovine muscle satellite cells1 J Anim Sci. 2019 97 4114 23 10.1093/jas/skz269 31424542

상세보기
15. Satory DL Smith SB Conjugated linoleic acid inhibits proliferation but stimulates lipid filling of murine 3T3-L1 preadipocytes J Nutr. 1999 129 92 7 10.1093/jn/129.1.92 9915881

상세보기
16. Deutsch MJ Schriever SC Roscher AA Ensenauer R Digital image analysis approach for lipid droplet size quantitation of Oil Red O-stained cultured cells Anal Biochem. 2014 445 87 9 10.1016/j.ab.2013.10.001 24120410

상세보기
17. Kook SH Choi KC Son YO Lee KY Hwang IH Lee HJ et al Satellite cells isolated from adult Hanwoo muscle can proliferate and differentiate into myoblasts and adipose-like cells Mol Cells 2006 22 239 45 17085978

상세보기
18. Xu J Liu D Yin H Tong H Li S Yan Y Fatty acids promote bovine skeletal muscle satellite cell differentiation by regulating ELOVL3 expression Cell Tissue Res. 2018 373 499 508 10.1007/s00441-018-2812-3 29464364

상세보기
19. Cassar-Malek I Langlois N Picard B Geay Y Regulation of bovine satellite cell proliferation and differentiation by insulin and triiodothyronine Domest Anim Endocrinol. 1999 17 373 88 10.1016/S0739-7240(99)00055-7 10628428

상세보기
20. Seale P Rudnicki MA A new look at the origin, function, and “stem-cell” status of muscle satellite cells Dev Biol. 2000 218 115 24 10.1006/dbio.1999.9565 10656756

상세보기
21. Han RH Wang M Fang X Han X Simulation of triacylglycerol ion profiles: bioinformatics for interpretation of triacylglycerol biosynthesis J Lipid Res. 2013 54 1023 32 10.1194/jlr.M033837 23365150

상세보기
22. Choi SH Chung KY Johnson BJ Go GW Kim KH Choi CW et al Co-culture of bovine muscle satellite cells with preadipocytes increases PPAR γ and C/EBP β gene expression in differentiated myoblasts and increases GPR43 gene expression in adipocytes J Nutr Biochem. 2013 24 539 43 10.1016/j.jnutbio.2012.01.015 22748806

상세보기
23. Choi SH Park SK Johnson BJ Chung KY Choi CW Kim KH et al AMPK α , C/EBP β , CPT1 β , GPR43, PPAR γ , and SCD gene expression in single- and co-cultured bovine satellite cells and intramuscular preadipocytes treated with palmitic, stearic, oleic, and linoleic acid Asian-australas J Anim Sci. 2015 28 411 9 10.5713/ajas.14.0598 25656188

원문보기 상세보기
24. Tontonoz P Hu E Spiegelman BM Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor γ Curr Opin Genet Dev. 1995 5 571 6 10.1016/0959-437X(95)80025-5 8664544

상세보기
25. Hollenberg AN Susulic VS Madura JP Zhang B Moller DE Tontonoz P et al Functional antagonism between CCAAT/Enhancer binding protein- α and peroxisome proliferator-activated receptor-gamma on the leptin promoter J Biol Chem. 1997 272 5283 90 10.1074/jbc.272.8.5283 9030601

상세보기
26. Kadegowda AKG Burns TA Pratt SL Duckett SK Inhibition of stearoyl-CoA desaturase 1 reduces lipogenesis in primary bovine adipocytes Lipids 2013 48 967 76 10.1007/s11745-013-3823-1 23929455

상세보기
27. Kim E Lee JH Ntambi JM Hyun CK Inhibition of stearoyl-CoA desaturase1 activates AMPK and exhibits beneficial lipid metabolic effects in vitro Eur J Pharmacol. 2011 672 38 44 10.1016/j.ejphar.2011.09.172 21970804

상세보기
28. Hardie DG Carling D The AMP-activated protein kinase--fuel gauge of the mammalian cell? Eur J Biochem. 1997 246 259 73 10.1111/j.1432-1033.1997.00259.x 9208914

상세보기
29. Bijland S Mancini SJ Salt IP Role of AMP-activated protein kinase in adipose tissue metabolism and inflammation Clin Sci. 2013 124 491 507 10.1042/CS20120536

상세보기
30. Fernandez-Veledo S Vazquez-Carballo A Vila-Bedmar R Ceperuelo-Mallafre V Vendrell J Role of energy- and nutrient-sensing kinases AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) in adipocyte differentiation IUBMB Life. 2013 65 572 83 10.1002/iub.1170 23671028

상세보기
31. Zong H Ren JM Young LH Pypaert M Mu J Birnbaum MJ et al AMP kinase is required for mitochondrial biogenesis in skeletal muscle in response to chronic energy deprivation Proc Nati Acad Sci USA 2002 99 15983 7 10.1073/pnas.252625599

상세보기
32. Bonen A Parolin ML Steinberg GR Calles-Escandon J Tandon NN Glatz JF et al Triacylglycerol accumulation in human obesity and type 2 diabetes is associated with increased rates of skeletal muscle fatty acid transport and increased sarcolemmal FAT/CD36 FASEB J. 2004 18 1144 6 10.1096/fj.03-1065fje 15132977

상세보기
33. Kelley DE Goodpaster B Wing RR Simoneau JA Skeletal muscle fatty acid metabolism in association with insulin resistance, obesity, and weight loss Am J Physiol. 1999 277 E1130 41 10.1152/ajpendo.1999.277.6.E1130 10600804

상세보기
34. Kelley DE Reilly JP Veneman T Mandarino LJ Effects of insulin on skeletal muscle glucose storage, oxidation, and glycolysis in humans Am J Physiol. 1990 258 E923 9 10.1152/ajpendo.1990.258.6.E923 2113769

상세보기
35. Simoneau JA Veerkamp JH Turcotte LP Kelley DE Markers of capacity to utilize fatty acids in human skeletal muscle: relation to insulin resistance and obesity and effects of weight loss FASEB J. 1999 13 2051 60 10.1096/fasebj.13.14.2051 10544188

상세보기
36. Chung KY Johnson BJ Melengestrol acetate enhances adipogenic gene expression in cultured muscle-derived cells J Anim Sci. 2009 87 3897 904 10.2527/jas.2008-1645 19684257

상세보기
37. Thiam AR Farese RV Jr. Walther TC The biophysics and cell biology of lipid droplets Nat Rev Mol Cell Biology. 2013 14 775 86 10.1038/nrm3699

상세보기
38. Bickel PE Tansey JT Welte MA PAT proteins, an ancient family of lipid droplet proteins that regulate cellular lipid stores Biochim Biophys Acta. 2009 1791 419 40 10.1016/j.bbalip.2009.04.002 19375517

상세보기
39. Greenberg AS Egan JJ Wek SA Garty NB Blanchette-Mackie EJ Londos C Perilipin, a major hormonally regulated adipocyte-specific phosphoprotein associated with the periphery of lipid storage droplets J Biol Chem. 1991 266 11341 6 10.1016/S0021-9258(18)99168-4 2040638

상세보기
40. Wolins NE Brasaemle DL Bickel PE A proposed model of fat packaging by exchangeable lipid droplet proteins FEBS Lett. 2006 580 5484 91 10.1016/j.febslet.2006.08.040 16962104

상세보기
41. Heid HW Moll R Schwetlick I Rackwitz HR Keenan TW Adipophilin is a specific marker of lipid accumulation in diverse cell types and diseases Cell Tiss Res. 1998 294 309 21 10.1007/s004410051181

상세보기
42. Brasaemle DL Barber T Wolins NE Serrero G Blanchette-Mackie EJ Londos C Adipose differentiation-related protein is an ubiquitously expressed lipid storage droplet-associated protein J Lipid Res. 1997 38 2249 63 10.1016/S0022-2275(20)34939-7 9392423

상세보기
43. Imamura M Inoguchi T Ikuyama S Taniguchi S Kobayashi K Nakashima N et al ADRP stimulates lipid accumulation and lipid droplet formation in murine fibroblasts Am J Physiol Endocrinol Metab. 2002 283 E775 83 10.1152/ajpendo.00040.2002 12217895

상세보기
44. Havel PJ Update on adipocyte hormones: regulation of energy balance and carbohydrate/lipid metabolism Diabetes 2004 53 Suppl S143 51 10.2337/diabetes.53.2007.S143

상세보기
45. Trayhurn P Endocrine and signalling role of adipose tissue: new perspectives on fat Acta Physiol Scand. 2005 184 285 93 10.1111/j.1365-201X.2005.01468.x 16026420

상세보기
46. Berg AH Combs TP Du X Brownlee M Scherer PE The adipocyte-secreted protein Acrp30 enhances hepatic insulin action Nat Med. 2001 7 947 53 10.1038/90992 11479628

상세보기
47. Ouchi N Kihara S Arita Y Maeda K Kuriyama H Okamoto Y et al Novel modulator for endothelial adhesion molecules: adipocyte-derived plasma protein adiponectin Circulation 1999 100 2473 6 10.1161/01.CIR.100.25.2473 10604883

상세보기
48. Yamauchi T Kamon J Waki H Terauchi Y Kubota N Hara K et al The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity Nat Med. 2001 7 941 6 10.1038/90984 11479627

상세보기
49. Christiaens V Van Hul M Lijnen HR Scroyen I CD36 promotes adipocyte differentiation and adipogenesis Biochim Biophys Acta. 2012 1820 949 56 10.1016/j.bbagen.2012.04.001 22507268

상세보기
50. Luiken JJ Chanda D Nabben M Neumann D Glatz JF Post-translational modifications of CD36 (SR-B2): implications for regulation of myocellular fatty acid uptake Biochim Biophys Acta. 2016 1862 2253 8 10.1016/j.bbadis.2016.09.004 27615427

상세보기

LOADING...

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