Ganglioside GM3 content in skeletal muscles is increased in type 2 but decreased in type 1 diabetes rat models: Implications of glycosphingolipid metabolism in pathophysiology of diabetes
骨骼肌中神经节苷脂GM3的含量在2型糖尿病大鼠模型中增加但在1型糖尿病大鼠模型中减少:鞘糖脂代谢在糖尿病病理生理学中的作用
Corresponding Author
Josko Bozic
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Correspondence
Josko Bozic, Department of Pathophysiology, University of Split School of Medicine, Soltanska 2, 21000 Split, Croatia.
Tel: +385 21 557 871
Fax: +385 21 557 895
Email: [email protected]
Search for more papers by this authorAnita Markotic
Department of Medical Chemistry and Biochemistry, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorVedrana Cikes-Culic
Department of Medical Chemistry and Biochemistry, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorAnela Novak
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorJosip A. Borovac
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorHrvoje Vucemilovic
Department of Anesthesiology and Intensive Care Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorGorana Trgo
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorTina Ticinovic Kurir
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorCorresponding Author
Josko Bozic
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Correspondence
Josko Bozic, Department of Pathophysiology, University of Split School of Medicine, Soltanska 2, 21000 Split, Croatia.
Tel: +385 21 557 871
Fax: +385 21 557 895
Email: [email protected]
Search for more papers by this authorAnita Markotic
Department of Medical Chemistry and Biochemistry, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorVedrana Cikes-Culic
Department of Medical Chemistry and Biochemistry, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorAnela Novak
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorJosip A. Borovac
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Search for more papers by this authorHrvoje Vucemilovic
Department of Anesthesiology and Intensive Care Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorGorana Trgo
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorTina Ticinovic Kurir
Department of Pathophysiology, University of Split School of Medicine, Split, Croatia
Department of Internal Medicine, University Hospital Split, Split, Croatia
Search for more papers by this authorAbstract
enBackground
Ganglioside GM3 is found in the plasma membrane, where its accumulation attenuates insulin receptor signaling. Considering the role of skeletal muscles in insulin-stimulated glucose uptake, the aim of the present study was to determine the expression of GM3 and its precursors in skeletal muscles of rat models of type 1 and type 2 diabetes mellitus (T1DM and T2DM, respectively).
Methods
Diabetes was induced in male Sprague-Dawley rats by streptozotocin injection (55 mg/kg, i.p., for T1DM induction; 35 mg/kg, i.p., for T2DM induction), followed by feeding of rats with either a normal pellet diet (T1DM) or a high-fat diet (T2DM). Rats were killed 2 weeks after diabetes induction and samples of skeletal muscle were collected. Frozen quadriceps muscle sections were stained with a primary antibody against GM3 (Neu5Ac) and visualized using a secondary antibody coupled with Texas Red. The muscle content of ganglioside GM3 and its precursors was analyzed by high-performance thin-layer chromatography (HPTLC) followed by GM3 immunostaining.
Results
Muscle GM3 content was significantly higher in T2DM compared with control rats (P < 0.001). Furthermore, levels of the GM3 precursors ceramide, glucosylceramide, and lactosylceramide were significantly higher in T2DM compared with control rats (P < 0.05), whereas ceramide content was significantly lower in T1DM rats (P < 0.05). The intensity of the GM3 band on HPTLC was significantly higher in T2DM rats (P < 0.001) and significantly lower in T1DM rats (P < 0.05) compared with control.
Conclusions
The expression patterns of GM3 ganglioside and its precursors in diabetic rats suggest that the role of glycosphingolipid metabolism may differ between T2DM and T1DM.
摘要
zh背景
神经节苷脂GM3在细胞膜上表达, 它在细胞膜上积聚可以减弱胰岛素受体的信号传导。考虑到骨骼肌在胰岛素刺激后的葡萄糖摄取中所起到的作用, 本研究旨在检测GM3及其前体在1型与2型糖尿病(分别缩写为T1DM与T2DM)大鼠模型骨骼肌中的表达。
方法
雄性Sprague-Dawley大鼠注射链脲霉素后诱导糖尿病(腹腔内注射55 mg/kg诱导T1DM;腹腔内注射35 mg/kg诱导T2DM), 然后分别用正常饲料(T1DM)或者高脂饲料(T2DM)进行饲养。诱导出糖尿病2周之后, 将大鼠处死并且收集骨骼肌样本进行实验。使用一级抗GM3抗体(Neu5Ac)对冰冻四头肌切片进行染色, 使用与德克萨斯红偶联的二级抗体显影。使用高效薄层色谱法以及GM3免疫染色法分析肌肉中神经节苷脂GM3及其前体含量。
结果
T2DM大鼠肌肉中的GM3含量显著高于对照组大鼠(P < 0.001)。此外, T2DM大鼠GM3前体神经酰胺、葡萄糖基神经酰胺以及乳糖基神经酰胺的水平也显著高于对照组大鼠(P < 0.05), 然而T1DM大鼠的神经酰胺含量却显著降低(P < 0.05)。与对照组大鼠相比, 使用高效薄层色谱法分析后发现T2DM大鼠的GM3条带强度显著增强(P < 0.001), 而T1DM大鼠中却显明显减弱(P < 0.05)。
结论
不同类型糖尿病大鼠的GM3神经节苷脂及其前体表达模式各不相同, 提示鞘糖脂代谢在T2DM与T1DM中的作用可能存在差异。
References
- 1 American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 2013; 36(Suppl. 1): S67–S74.
- 2 Kaur J. A comprehensive review on metabolic syndrome. Cardiol Res Pract. 2014; 2014: 943162.
- 3 Taylor R. Type 2 diabetes etiology and reversibility. Diabetes Care. 2013; 36: 1047–1055.
- 4 Ferrante A. Obesity-induced inflammation: A metabolic dialogue in the language of inflammation. J Intern Med. 2007; 262: 408–414.
- 5 Alberti KG, Zimmet P, Shaw J. Metabolic syndrome: A new world-wide definition. A Consensus Statement from the International Diabetes Federation. Diabet Med. 2006; 23: 469–480.
- 6 Mozaffarian D, Benjamin EJ, Go AS et al. Heart disease and stroke statistics – 2016 update: A report from the American Heart Association. Circulation. 2016; 133: e38–e360.
- 7 Semenkovich CF. Insulin resistance and a long, strange trip. N Engl J Med. 2016; 374: 1378–1379.
- 8 Van Greevenbroek M, Schalkwijk C, Stehouwer C. Obesity-associated low-grade inflammation in type 2 diabetes mellitus: Causes and consequences. Neth J Med. 2013; 71: 174–187.
- 9 Carlsson AC, Östgren CJ, Nystrom FH et al. Association of soluble tumor necrosis factor receptors 1 and 2 with nephropathy, cardiovascular events, and total mortality in type 2 diabetes. Cardiovasc Diabetol. 2016; 15: 1–8.
- 10 Lee YS, Li P, Huh JY et al. Inflammation is necessary for long-term but not short-term high-fat diet-induced insulin resistance. Diabetes. 2011; 60: 2474–2483.
- 11 Olefsky JM, Glass CK. Macrophages, inflammation, and insulin resistance. Annu Rev Physiol. 2010; 72: 219–246.
- 12 Kabayama K, Sato T, Kitamura F et al. TNFα-induced insulin resistance in adipocytes as a membrane microdomain disorder: Involvement of ganglioside GM3. Glycobiology. 2005; 15: 21–29.
- 13 Vukovic I, Bozic J, Markotic A, Ljubicic S, Ticinovic Kurir T. The missing link: Likely pathogenetic role of GM3 and other gangliosides in the development of diabetic nephropathy. Kidney Blood Press Res. 2015; 40: 306–314.
- 14 Lipina C, Hundal HS. Ganglioside GM3 as a gatekeeper of obesity-associated insulin resistance: Evidence and mechanisms. FEBS Lett. 2015; 589: 3221–3227.
- 15 Bergman BC, Brozinick JT, Strauss A et al. Serum sphingolipids: Relationships to insulin sensitivity and changes with exercise in humans. Am J Physiol Endocrinol Metab. 2015; 309: E398–E408.
- 16 Tagami S, Inokuchi J, Kabayama K et al. Ganglioside GM3 participates in the pathological conditions of insulin resistance. J Biol Chem. 2002; 277: 3085–3092.
- 17 Yamashita T, Hashiramoto A, Haluzik M et al. Enhanced insulin sensitivity in mice lacking ganglioside GM3. Proc Natl Acad Sci USA. 2003; 100: 3445–3449.
- 18 Inokuchi J. Physiopathological function of hematoside (GM3 ganglioside). Proc Jpn Acad Ser B. 2011; 87: 179–198.
- 19 Randeria PS, Seeger MA, Wang XQ et al. siRNA-based spherical nucleic acids reverse impaired wound healing in diabetic mice by ganglioside GM3 synthase knockdown. Proc Natl Acad Sci USA. 2015; 112: 5573–5578.
- 20 Sandhoff K, Kolter T. Biosynthesis and degradation of mammalian glycosphingolipids. Philos Trans R Soc Lond B Biol Sci. 2003; 358: 847–861.
- 21 Novak A, Rezic Muzinic N, Cikes Culic V et al. Renal distribution of ganglioside GM3 in rat models of types 1 and 2 diabetes. J Physiol Biochem. 2013; 69: 727–735.
- 22 Wolff RA, Dobrowsky RT, Bielawska A, Obeid LM, Hannun YA. Role of ceramide-activated protein phosphatase in ceramide-mediated signal transduction. J Biol Chem. 1994; 269: 19605–19609.
- 23 Kowluru A, Metz SA. Ceramide-activated protein phosphatase-2A activity in insulin-secreting cells. FEBS Lett. 1997; 418: 179–182.
- 24 Chalfant CE, Kishikawa K, Mumby MC, Kamibayashi C, Bielawska A, Hannun YA. Long chain ceramides activate protein phosphatase-1 and protein phosphatase-2A. Activation is stereospecific and regulated by phosphatidic acid. J Biol Chem. 1999; 274: 20313–20317.
- 25 Chakraborty A, Koldobskiy MA, Bello NT et al. Inositol pyrophosphates inhibit Akt signaling, thereby regulating insulin sensitivity and weight gain. Cell. 2010; 143: 897–910.
- 26 Sriwijitkamol A, Coletta DK, Wajcberg E et al. Effect of acute exercise on AMPK signaling in skeletal muscle of subjects with type 2 diabetes: A time-course and dose-response study. Diabetes. 2007; 56: 836–848.
- 27
National Research Council Committee for the Update of the Guide for the Care and Use of Laboratory Animals. Guide for the Care and Use of Laboratory Animals. The National Academies Collection: Reports funded by National Institutes of Health. National Academies Press (US) National Academy of Sciences, Washington DC, 2011.
10.17226/25801 Google Scholar
- 28 Rerup CC. Drugs producing diabetes through damage of the insulin secreting cells. Pharmacol Rev. 1970; 22: 485–518.
- 29 Srinivasan K, Viswanad B, Asrat L, Kaul C, Ramarao P. Combination of high-fat diet-fed and low-dose streptozotocin-treated rat: A model for type 2 diabetes and pharmacological screening. Pharmacol Res. 2005; 52: 313–320.
- 30 Lee G, Goosens KA. Sampling blood from the lateral tail vein of the rat. J Vis Exp. 2015: e52766. https://doi.org/10.3791/52766.
- 31 Steil D, Schepers CL, Pohlentz G et al. Shiga toxin glycosphingolipid receptors of Vero-B4 kidney epithelial cells and their membrane microdomain lipid environment. J Lipid Res. 2015; 56: 2322–2336.
- 32 Rešić J, Čikeš Čulić V, Zemunik T, Markotić A. Hepatic and pancreatic glycosphingolipid phenotypes of the neurological different rat strains. Coll Antropol. 2011; 35: 259–263.
- 33 Ticinovic-Kurir T, Cikes-Culic V, Zemunik T et al. Immunohistochemical analysis of hepatic ganglioside distribution following a partial hepatectomy and exposure to different hyperbaric oxygen treatments. Acta Histochem. 2008; 110: 66–75.
- 34 Markotic A, Culic VC, Kurir TT et al. Oxygenation alters ganglioside expression in rat liver following partial hepatectomy. Biochem Biophys Res Commun. 2005; 330: 131–141.
- 35 Aljada A, Mohanty P, Ghanim H et al. Increase in intranuclear nuclear factor κB and decrease in inhibitor κB in mononuclear cells after a mixed meal: Evidence for a proinflammatory effect. Am J Clin Nutr. 2004; 79: 682–690.
- 36
Chatterjee S, Mishra S, Suzuki SK. New vis-tas in lactosylceramide research. In: A Chakrabarti, A Surolia (eds). Biochemical Roles of Eukaryotic Cell Surface Macromolecules. Springer International Publishing, Cham, 2015; 127–138.
10.1007/978-3-319-11280-0_8 Google Scholar
- 37 Nakamura H, Moriyama Y, Makiyama T et al. Lactosylceramide interacts with and activates cytosolic phospholipase A2α. J Biol Chem. 2013; 288: 23 264–23 272.
- 38 Erdei N, Bagi Z, Édes I, Kaley G, Koller A. H2O2 increases production of constrictor prostaglandins in smooth muscle leading to enhanced arteriolar tone in type 2 diabetic mice. Am J Physiol Heart Circ Physiol. 2007; 292: H649–H656.
- 39 Natarajan R, Nadler JL. Lipid inflammatory mediators in diabetic vascular disease. Arterioscler Thromb Vasc Biol. 2004; 24: 1542–1548.
- 40 Obanda DN, Yu Y, Wang ZQ, Cefalu WT. Modulation of sphingolipid metabolism with calorie restriction enhances insulin action in skeletal muscle. J Nutr Biochem. 2015; 26: 687–695.
- 41 Bruce CR, Risis S, Babb JR et al. The sphingosine-1-phosphate analog FTY720 reduces muscle ceramide content and improves glucose tolerance in high fat-fed male mice. Endocrinology. 2012; 154: 65–76.
- 42 Kitada Y, Kajita K, Taguchi K et al. Blockade of sphingosine 1-phosphate receptor 2 signaling attenuates high-fat diet-induced adipocyte hypertrophy and systemic glucose intolerance in mice. Endocrinology. 2016; 157: 1839–1851.
- 43 Lafontan M, Langin D. Lipolysis and lipid mobilization in human adipose tissue. Prog Lipid Res. 2009; 48: 275–297.
- 44 Bergman BC, Cornier M-A, Horton TJ, Bessesen DH. Effects of fasting on insulin action and glucose kinetics in lean and obese men and women. Am J Physiol Endocrinol Metab. 2007; 293: E1103–E1111.
- 45 Fox TE, Bewley MC, Unrath KA et al. Circulating sphingolipid biomarkers in models of type 1 diabetes. J Lipid Res. 2011; 52: 509–517.
- 46 Chavez JA, Siddique MM, Wang ST, Ching J, Shayman JA, Summers SA. Ceramides and glucosylceramides are independent antagonists of insulin signaling. J Biol Chem. 2014; 289: 723–734.
- 47 Hage Hassan R, Pacheco de Sousa AC, Mahfouz R et al. Sustained action of ceramide on the insulin signaling pathway in muscle cells: Implication of the double-stranded RNA-activated protein kinase. J Biol Chem. 2016; 291: 3019–3029.
- 48 Baird TD, Wek RC. Eukaryotic initiation factor 2 phosphorylation and translational control in metabolism. Adv Nutr (Bethesda, Md). 2012; 3: 307–321.
- 49 Carvalho BM, Oliveira AG, Ueno M et al. Modulation of double-stranded RNA-activated protein kinase in insulin sensitive tissues of obese humans. Obesity (Silver Spring, Md). 2013; 21: 2452–2457.
- 50 Camell CD, Nguyen KY, Jurczak MJ et al. Macrophage-specific de novo synthesis of ceramide is dispensable for inflammasome-driven inflammation and insulin resistance in obesity. J Biol Chem. 2015; 290: 29402–29413.
- 51 Boden G. Obesity, insulin resistance and free fatty acids. Curr Opin Endocrinol Diabetes Obes. 2011; 18: 139–143.
- 52 Frohnert BI, Jacobs DR, Steinberger J, Moran A, Steffen LM, Sinaiko AR. Relation between serum free fatty acids and adiposity, insulin resistance, and cardiovascular risk factors from adolescence to adulthood. Diabetes. 2013; 62: 3163.
- 53 Schmitz-Peiffer C, Craig DL, Biden TJ. Ceramide generation is sufficient to account for the inhibition of the insulin-stimulated PKB pathway in C2C12 skeletal muscle cells pretreated with palmitate. J Biol Chem. 1999; 274: 24202–24210.
- 54 Ghosh N, Patel N, Jiang K et al. Ceramide-activated protein phosphatase (CAPP) involvement in insulin resistance via Akt, SRp40, and RNA splicing in L6 skeletal muscle cells. Endocrinology. 2007; 148: 1359–1366.
- 55 Stratford S, Hoehn KL, Liu F, Summers SA. Regulation of insulin action by ceramide dual mechanisms linking ceramide accumulation to the inhibition of Akt/protein kinase B. J Biol Chem. 2004; 279: 36608–36615.
- 56 Kabayama K, Sato T, Saito K et al. Dissociation of the insulin receptor and caveolin-1 complex by ganglioside GM3 in the state of insulin resistance. Proc Natl Acad Sci USA. 2007; 104: 13678–13683.
- 57 Catalán V, Gómez-Ambrosi J, Rodríguez A et al. Expression of caveolin-1 in human adipose tissue is upregulated in obesity and obesity-associated type 2 diabetes mellitus and related to inflammation. Clin Endocrinol. 2008; 68: 213–219.
- 58 Couet J, Li S, Okamoto T, Ikezu T, Lisanti MP. Identification of peptide and protein ligands for the caveolin-scaffolding domain: Implications for the interaction of caveolin with caveolae-associated proteins. J Biol Chem. 1997; 272: 6525–6533.
- 59 Cohen AW, Razani B, Wang XB et al. Caveolin-1-deficient mice show insulin resistance and defective insulin receptor protein expression in adipose tissue. Am J Physiol Cell Physiol. 2003; 285: C222–C235.
- 60 Williams TM, Lisanti MP. The caveolin proteins. Genome Biol. 2004; 5: 214.
- 61 Wang H, Wang AX, Aylor K, Barrett EJ. Caveolin-1 phosphorylation regulates vascular endothelial insulin uptake and is impaired by insulin resistance in rats. Diabetologia. 2015; 58: 1344–1353.
- 62 Tang Z, Scherer PE, Okamoto T et al. Molecular cloning of caveolin-3, a novel member of the caveolin gene family expressed predominantly in muscle. J Biol Chem. 1996; 271: 2255–2261.
- 63 Capozza F, Combs TP, Cohen AW et al. Caveolin-3 knockout mice show increased adiposity and whole body insulin resistance, with ligand-induced insulin receptor instability in skeletal muscle. Am J Physiol Cell Physiol. 2005; 288: C1317–C1331.
- 64 Bruni P, Donati C. Pleiotropic effects of sphingolipids in skeletal muscle. Cell Mol Life Sci. 2008; 65: 3725–3736.
- 65 Tonks KT, Coster ACF, Christopher MJ et al. Skeletal muscle and plasma lipidomic signatures of insulin resistance and overweight/obesity in humans. Obesity. 2016; 24: 908–916.
- 66 Zhao H, Przybylska M, I-H W et al. Inhibiting glycosphingolipid synthesis improves glycemic control and insulin sensitivity in animal models of type 2 diabetes. Diabetes. 2007; 56: 1210–1218.
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