Effect of High-fat or High-glucose Diet on Obesity and Visceral Adipose Tissue in Mice
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1 ACTA ACADEMIAE MEDICINAE SINICAE com 4 C57BL / mrna 20 P < O 20 P > M2 P < mrna P < P > R DOI /j. issn X A X 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 China Corresponding author LI Xia Tel lixia2014@vip 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 < 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 Supported by the National Natural Sciences Foundation of China The first two author contributed equally to this article 614
2 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 < Compared with normal diet group monocyte chemoattractant protein-1 mrna expression increased significantly in high-fat diet group P < In highglucose group however no significant differences were discerned P > 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 mm 2 mm 1 C6885 Sigma 2 USA min -1 monocyte chemoattractant protein-1 MCP cm r /min 5 min M1 M2 M1 Th1 M2 Millipore Th2 Excel 5 10 μm 35 C57BL /6 10 min O 5 min F4 /80 CD11c CD11b 5 20 Biolengend Flow Cytometer F4 /80 + MCP-1 mrna 60% CD11b + CD11c + M1 F4 / % CD11b + CD11c - M2 10% MCP-1 mrna Trizol RNA cdna MCP-1 5 -CACT- CACCTGCTGCTACTCAT-3 5 -CTACAGCTTCTTTGGGA
3 CACC-3 PCR ABI- PRISM 7000 PCR threshold cycle val- PCR ± g ± g ± g PCR ± g P < ue CT ± g ± g SPSS ± g P = ± P = A 1B ± % ± % ONE-WAY ANOVA P < ±0. 60 % ± P < % P = C ± ng /ml ± ng /ml P = ± g ± g ± g ± g P < P < ± g ± g ± ng /ml ± ng /ml ± g ± g P < ± g ± ng /ml ± ng /ml P < ± ng /ml ± ng /ml P = D ND HG HF a P < ND normal diet group HG high-glucose diet group HF high-fat diet group a P < 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
4 O ± % ± % 3C M ± % ± % 10 P < D ± % ± % A. B. C. A. normal diet group B. high-fat diet group C. high-glucose diet group 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 < FITC fluorescein isothiocyanate APC allophycocyanin FSC forward scattering angle PerCP peridinin-chlorophyll-protein M1 M1 macrophages M2 M2 macrophages a P < 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
5 MCP-1 mrna PCR 10 60% MCP-1 20 MCP ± ± P < 60% MCP ± ± P > α tumor necrosis factorα TNF-α interleukin IL -6 IL-1β M1 IL-10 β M MCP-1 mrna 20 4 Fig 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 < MCP-1 monocyte chemoattractant protein-1 a P < compared with Huang 8 the normal diet group NADPH NADPH 618
6 Matos Ferreira 14 Swiss 64% 19% 7 Chinetti-Gbaguidi G Staels B. 11% 45% 17% 38% Lipidol MCP-1 MCP-1 Scientific World J 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 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 / Cipolletta D. Adipose tissue-resident regulatory T cells phenotypic specialization functions and therapeutic potential J. Immunology 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 e Deshmane SL Kremlev S Amini S et al. Monocyte chemoattractant protein-1 MCP-1 an overview J. J Interferon Cytokine Res 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 Sica A Mantovani A. Macrophage plasticity and polarization in vivo veritas J. J Clin Invest Lee 13 C57BL / Weisberg SP Mccann D Desai M et al. Obesity is associ- TNF-α ated with macrophage accumulation in adipose tissue J. J Clin Invest 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 Makki K Froguel P Wolowczuk I. Adipose tissue in obesityrelated inflammation and insulin resistance cells cytokines and chemokines J. ISRN Inflamm 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 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
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