ACTA ACADEMIAE MEDICINAE SINICAE 410011 0731-85295846 lixia2014@vip. 163. com 4 C57BL /6 20-1 mrna 20 P < 0. 05 O 20 P > 0. 05 M2 P < 0. 05-1 mrna P < 0. 05 P > 0. 05 R589. 2 DOI 10. 3881 /j. issn. 1000-503X. 2014. 06. 010 A 1000-503X 2014 06-0614-06 Effect of High-fat or High-glucose Diet on Obesity and Visceral Adipose Tissue in Mice TANG Li-li TANG Xiao-han LI Xia YU Hai-bo XIE Zhi-guo LIU Xin-yuan ZHOU Zhi-guang Department of Endocrinology the Second Xiangya Hospital Central South University Diabetes Center Institute of Metabolism and Endocrinology Central South University Diabetes Immunology Ministry of Education Key Laboratory National Clinical Research Center for Metabolic Disease Changsha 410011 China Corresponding author LI Xia Tel 0731-85295846 E-mail lixia2014@vip. 163. com ABSTRACT Objective To investigate the effect of high-fat or high-glucose diet on obesity and visceral adipose tissue in C57BL /6 mice. Methods Four-week-old C57BL /6 mice were allocated into normal diet group high-fat diet group and high-glucose diet group according to the random number table until 20 weeks old. Body weight epididymal adipose tissue weight blood leptin fat infiltration in liver M1 /M2 macrophage subtypes and monocyte chemoattractant protein-1 mrna in epididymal adipose tissues were measured. Results Compared with normal diet group body weight epididymal adipose tissue weight and leptin concentration in high fat diet group at 20 weeks were significantly increased P < 0. 05 and oil red O staining showed more prominent adipocyte infiltration in liver in high-fat diet group than those in normal diet and high-glucose diet 81070672 Supported by the National Natural Sciences Foundation of China 81070672 The first two author contributed equally to this article 614
group. However no apparent differences were seen in high-glucose diet group at 20 weeks in terms of body weight epididymal adipose tissue weight and leptin concentration. In high-fat diet group the macrophages infiltration in epididymal adipose tissue increased with time and the percentage of M2 macrophage decreased in highfat diet group than that in high-glucose diet group P < 0. 05. Compared with normal diet group monocyte chemoattractant protein-1 mrna expression increased significantly in high-fat diet group P < 0. 05. In highglucose group however no significant differences were discerned P > 0. 05. Conclusion High-fat diet rather than 60% high glucose diet will lead to obesity and macrophage infiltration in adipose tissues. Key words adipose tissue high-fat diet high-glucose diet macrophages obesity Acta Acad Med Sin 2014 36 6 614-619 2 mm 2 mm 1 C6885 Sigma 2 USA 37 30 min -1 monocyte chemoattractant protein-1 MCP-1 11. 5 cm 4 1500 r /min 5 min 3 3 4 M1 M2 M1 Th1 M2 Millipore Th2 Excel 5 10 μm 35 C57BL /6 10 min O 5 min 3 4 10 4 5 F4 /80 CD11c 2 5 10 CD11b 5 20 Biolengend Flow Cytometer F4 /80 + MCP-1 mrna 60% CD11b + CD11c + M1 F4 /80 + 60% CD11b + CD11c - M2 10% MCP-1 mrna Trizol RNA cdna MCP-1 5 -CACT- CACCTGCTGCTACTCAT-3 5 -CTACAGCTTCTTTGGGA-.. 615
CACC-3 PCR ABI- PRISM 7000 PCR threshold cycle val- PCR 20 39. 05 ± 2. 33 g 31. 65 ± 1. 20 g 2. 12 ± 0. 48 g PCR 0. 36 ± 0. 09 g P < 0. 05 20 ue CT 3 31. 65 ± 1. 20 g 0. 42 ± 0. 19 g SPSS 17. 0 0. 36 ± 0. 09 g P = 0. 26 ± P = 0. 60 1A 1B 20 5. 39 ± 0. 89 % 1. 14 ± 0. 28 % ONE-WAY ANOVA P <0. 05 1. 38 ±0. 60 % 1. 14 ± P < 0. 05 0. 28 % P = 0. 60 1C 2. 55 ± 0. 85 ng /ml 1. 21 ± 10 10 0. 45 ng /ml P = 0. 06 30. 40 ± 0. 88 g 24. 04 ± 1. 11 g 20 0. 75 ± 0. 12 g 0. 32 ± 0. 11 g P < 0. 05 10 P < 0. 05 26. 18 ± 0. 93 g 24. 04 ± 1. 11 g 3. 10 ± 1. 64 ng /ml 2. 14 ± 0. 60 ng /ml 0. 52 ± 0. 03 g 0. 32 ± 0. 11 g P < 0. 05 20 30. 05 ± 1. 95 g 10 4. 52 ± 1. 47 ng /ml 1. 21 ± 0. 45 ng /ml P < 0. 05 13. 57 ± 3. 44 ng /ml 2. 14 ± 0. 60 ng /ml P = 0. 56 1D ND HG HF a P < 0. 05 ND normal diet group HG high-glucose diet group HF high-fat diet group a P < 0. 05 compared with the normal diet group A. B. C. D. A. body weight B. epididymal adipose tissue weight C. percentage of epididymal adipose tissue weight ratio D. serum leptin levels 1 Fig 1 Comparison of body weight epididymal adipose tissue weight and leptin levels among three groups 616
O 20 20 10 4. 56 ± 2 3. 00 % 20 16. 20 ± 3. 80 % 3C M2 10 20 8. 36 ± 7. 54 % 21. 15 ± 11. 22 % 10 P < 0. 05 3D 54. 73 ± 4. 47 % 70. 57 ± 12. 55 % A. B. C. A. normal diet group B. high-fat diet group C. high-glucose diet group 2 20 20 Fig 2 Comparison of adipocyte infiltration among three groups by liver oil red staining 20 FITC APC FSC PerCP - - M1 M1 M2 M2 a P < 0. 05 FITC fluorescein isothiocyanate APC allophycocyanin FSC forward scattering angle PerCP peridinin-chlorophyll-protein M1 M1 macrophages M2 M2 macrophages a P < 0. 05 compared with the high-glucose diet group A. B. M1 C. D. 20 M1 /M2 A. flow chart of macrophages in adipose tissue in high-fat diet group B. flow chart of M1 type macrophages in adipose tissue in high-fat diet group C. comparison of percentage of macrophages for each group D. comparison of percentage of M2 macrophage at week 20 for each group 3 Fig 3 Comparison of macrophages infiltration among three groups.. 617
MCP-1 mrna PCR 10 60% MCP-1 20 MCP-1 10. 96 ± 6. 32 1. 50 ± 0. 82 P < 60% 0. 05 MCP-1 2. 36 ± 2. 92 1. 50 ± 0. 82 P > 0. 05 4 2 10 20 6 -α tumor necrosis factorα TNF-α interleukin IL -6 IL-1β M1 IL-10 β M2 7 10 MCP-1 mrna 20 4 Fig 4 4 10 20 MCP-1 mrna Expressions of MCP-1 mrna at the 4 th 10 th and 20 th week in each group MCP-1 MCP-1 M2 M2 M1 MCP-1-1 a P < 0. 05 MCP-1 monocyte chemoattractant protein-1 a P < 0. 05 compared with Huang 8 the normal diet group NADPH NADPH 618
Matos Ferreira 14 Swiss 64% 19% 7 Chinetti-Gbaguidi G Staels B. 11% 45% 17% 38% Lipidol 2011 22 5 365-372. MCP-1 MCP-1 Scientific World J 2013 2013 752071. 9 Dickinson S Hancock DP Petocz P et al. High-glycemic 20 MCP-1 in mononuclear cells of young lean healthy subjects J. 10 Ghanim H Aljada A Hofmeyer D et al. Circulating mono- 60% nuclear cells in the obese are in a proinflammatory state J. Circulation 2004 110 12 1564-1571. 11 Van der Heiden K Cuhlmann S Luong LA et al. Role of nuclear factor kappa B in cardiovascular health and disease J. Clin Sci Lond 2010 118 9 /10 593-605. 1 Cipolletta D. Adipose tissue-resident regulatory T cells phenotypic specialization functions and therapeutic potential J. Immunology 2014 142 4 517-525. 2 Jakobsdottir G Xu J Molin G et al. High-fat diet reduces the formation of butyrate but increases succinate inflammation liver fat and cholesterol in rats while dietary fibre counteracts these effects J. PLoS One 2013 8 11 e80476. 3 Deshmane SL Kremlev S Amini S et al. Monocyte chemoattractant protein-1 MCP-1 an overview J. J Interferon Cytokine Res 2009 29 6 313-326. 4 Kanda H Tateya S Tamori Y et al. MCP-1 contributes to -κb TNF-α 9-11 MCP-1 macrophage infiltration into adipose tissue insulin resistance and hepatic steatosis in obesity J. J Clin Invest M1 TNF-α IL-6 2006 116 6 1494-1505. 12 5 Sica A Mantovani A. Macrophage plasticity and polarization in vivo veritas J. J Clin Invest 2012 122 3 Lee 13 C57BL /6 787-795. 6 Weisberg SP Mccann D Desai M et al. Obesity is associ- TNF-α ated with macrophage accumulation in adipose tissue J. J Clin Invest 2003 112 12 1796-1808. Macrophage polarization in metabolic disorders functions and regulation J. Curr Opin 8 Huang CJ Zourdos MC Jo E et al. Influence of physical activity and nutrition on obesity-related immune function J. index carbohydrate increases nuclear factor-kappa B activation Am J Clin Nutr 2008 87 5 1188-1193. 12 Makki K Froguel P Wolowczuk I. Adipose tissue in obesityrelated inflammation and insulin resistance cells cytokines and chemokines J. ISRN Inflamm 2013 2013 139239. 13 Lee IS Shin G Choue R. Shifts in diet from high fat to high carbohydrate lmproved levels of adipokines and pro-inflammatory cytokines in mice fed a high-fat diet J. Endocr J 2010 57 1 39-50. 14 Matos Ferreira AV Mario EG Jardim Porto LC et al. High-carbohydrate diet selectively induces tumor necrosis factor-alpha production in mice liver J. Inflammation 2011 34 2 139-145. 2014-04-21.. 619