Similar articles

Abstract: This study compared the effects on bone metabolism and morphology of pathological obesity induced by excessive fat intake in a non-hibernator (mice) versus healthy obesity due to pre-hibernation fattening in a hibernator (ground squirrels). Kunming mice were fed a high-fat diet to provide a model of pathological obesity (OB group). Daurian ground squirrels fattened naturally in their pre-hibernation season (PRE group) were used as a healthy obesity model. Micro-computed tomography (micro-CT) and three-point bending tests were used to determine the microstructure and mechanical properties of bone. Western blots were used to analyze protein expression levels related to bone metabolism (Runt-related transcription factor 2 (RunX2), osteocalcin (OCN), alkaline phosphatase (ALP), osteoprotegerin (OPG), receptor activator of nuclear factor-‍κB ligand (RANKL), cathepsin K, matrix metallopeptidase 9 (MMP9), patched protein homolog 1 (Ptch1), phosphorylated β‍-‍catenin (P‍-‍β‍-‍catenin), and glycogen synthase kinase-3β(GSK-3β)). Compared with controls, there was no obvious bone loss in the OB mice, and the stiffness of the femur was increased significantly. Compared with summer active squirrels, bone formation was enhanced but the mechanical properties did not change in the PRE group squirrels. In OB mice, western blots showed significantly increased expression levels of all proteins except RunX2, OPG, and Ptch1. PRE ground squirrels showed significantly increased expression of most proteins except OCN and Ptch1, which decreased significantly, and P‍-‍β‍-‍catenin and OPG, which did not change. In conclusion, for non-hibernating mice, moderate obesity had a certain protective effect on bones, demonstrating two-way regulation, increasing both bone loss and bone formation. For pre-hibernating ground squirrels, the healthy obesity acquired before hibernation had a positive effect on the microstructure of bones, and also enhanced the expression levels of proteins related to bone formation, bone resorption, and wnt signaling.

高脂饮食小鼠和冬眠前自然育肥达乌尔黄鼠的骨代谢和蛋白质表达差异

[1]BaronR, KneisselM, 2013. Wnt signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med, 19(2):179-192.

[2]BettisT, KimBJ, HamrickMW, 2018. Impact of muscle atrophy on bone metabolism and bone strength: implications for muscle-bone crosstalk with aging and disuse. Osteoporos Int, 29(8):1713-1720.

[3]BhattaraiT, BhattacharyaK, ChaudhuriP, et al., 2014. Correlation of common biochemical markers for bone turnover, serum calcium, and alkaline phosphatase in post-menopausal women. Malays J Med Sci, 21(1):58-61.

[4]BuchetR, MillánJL, MagneD, 2013. Multisystemic functions of alkaline phosphatases. In: Millán J (Ed.), Phosphatase Modulators. Methods in Molecular Biology, Vol. 1053. PressHumana, Totowa, NJ, p.27-51.

[5]BusutilR, EspallardoO, TorresA, et al., 2017. The impact of obesity on health-related quality of life in Spain. Health Qual Life Outcomes, 15:197.

[6]CaoJJ, PickloMJ, 2014. N-Acetylcysteine supplementation decreases osteoclast differentiation and increases bone mass in mice fed a high-fat diet. J Nutr, 144(3):289-296.

[7]CaoJJ, GregoireBR, MichelsenKG, et al., 2020. Increasing dietary fish oil reduces adiposity and mitigates bone deterioration in growing C57BL/6 mice fed a high-fat diet. J Nutr, 150(1):99-107.

[8]ChenFL, WangY, WangHW, et al., 2019. Flaxseed oil ameliorated high-fat-diet-induced bone loss in rats by promoting osteoblastic function in rat primary osteoblasts. Nutr Metab (Lond), 16:71.

[9]ChenHL, LiJF, WangQ, 2018. Associations between bone-alkaline phosphatase and bone mineral density in adults with and without diabetes. Medicine (Baltimore), 97(17):e0432.

[10]ChenJR, LazarenkoOP, WuXL, et al., 2010. Obesity reduces bone density associated with activation of PPARγ and suppression of Wnt/β-catenin in rapidly growing male rats. PLoS ONE, 5(10):e13704.

[11]ChungAWY, HsiangYN, MatzkeLA, et al., 2006. Reduced expression of vascular endothelial growth factor paralleled with the increased angiostatin expression resulting from the upregulated activities of matrix metalloproteinase-2 and -9 in human type 2 diabetic arterial vasculature. Circ Res, 99(2):140-148.

[12]ClarkEM, NessAR, TobiasJH, 2006. Adipose tissue stimulates bone growth in prepubertal children. J Clin Endocrinol Metab, 91(7):2534-2541.

[13]DallasSL, PrideauxM, BonewaldLF, 2013. The osteocyte: an endocrine cell ... and more. Endocr Rev, 34(5):658-690.

[14]DattaHK, NgWF, WalkerJA, et al., 2008. The cell biology of bone metabolism. J Clin Pathol, 61(5):577-587.

[15]DayTF, YangYZ, 2008. Wnt and hedgehog signaling pathways in bone development. J Bone Joint Surg Am, 90(Suppl 1):19-24.

[16]DucyP, DesboisC, BoyceB, et al., 1996. Increased bone formation in osteocalcin-deficient mice. Nature, 382(6590):448-452.

[17]FangZY, LinR, YuanBX, et al., 2007. Tanshinone IIA inhibits atherosclerotic plaque formation by down-regulating MMP-2 and MMP-9 expression in rabbits fed a high-fat diet. Life Sci, 81(17-18):1339-1345.

[18]FengYM, WanPB, YinLL, et al., 2020. The inhibition of microRNA-139-5p promoted osteoporosis of bone marrow-derived mesenchymal stem cells by targeting Wnt/beta-catenin signaling pathway by NOTCH1. J Microbiol Biotechnol, 30(3):448-458.

[19]GaoXL, WangSQ, ZhangJ, et al., 2021. Differential bone remodeling mechanism in hindlimb unloaded rats and hibernating Daurian ground squirrels: a comparison between artificial and natural disuse. J Comp Physiol B, 191(4):793-814.

[20]GautamJ, ChoudharyD, KhedgikarV, et al., 2014. Micro-architectural changes in cancellous bone differ in female and male C57BL/6 mice with high-fat diet-induced low bone mineral density. Br J Nutr, 111(10):1811-1821.

[21]GlassDA II, BialekP, AhnJD, et al., 2005. Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation. Dev Cell, 8(5):751-764.

[22]GongYQ, SleeRB, FukaiN, et al., 2001. LDL receptor-related protein 5 (LRP5) affects bone accrual and eye development. Cell, 107(4):513-523.

[23]GuY, WangZR, ShiJW, et al., 2017. Titanium particle-induced osteogenic inhibition and bone destruction are mediated by the GSK-3β/β‍-catenin signal pathway. Cell Death Dis, 8(6):e2878.

[24]HaladeGV, RahmanM, WilliamsPJ, et al., 2010. High fat diet-induced animal model of age-associated obesity and osteoporosis. J Nutr Biochem, 21(12):1162-1169.

[25]HaladeGV, el JamaliA, WilliamsPJ, et al., 2011. Obesity-mediated inflammatory microenvironment stimulates osteoclastogenesis and bone loss in mice. Exp Gerontol, 46(1):43-52.

[26]HaladeGV, JinYF, LindseyML, 2013. Matrix metalloproteinase (MMP)‍-9: a proximal biomarker for cardiac remodeling and a distal biomarker for inflammation. Pharmacol Ther, 139(1):32-40.

[27]HillTP, SpäterD, TaketoMM, et al., 2005. Canonical Wnt/β‍-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell, 8(5):727-738.

[28]HoeppnerLH, SecretoFJ, WestendorfJJ, 2009. Wnt signaling as a therapeutic target for bone diseases. Expert Opin Ther Targets, 13(4):485-496.

[29]HwangJH, ChaPH, HanG, et al., 2015. Euodia sutchuenensis dode extract stimulates osteoblast differentiation via Wnt/β‍-catenin pathway activation. Exp Mol Med, 47(3):e152.

[30]Ionova-MartinSS, DoSH, BarthHD, et al., 2010. Reduced size-independent mechanical properties of cortical bone in high-fat diet-induced obesity. Bone, 46(1):217-225.

[31]Ionova-MartinSS, WadeJM, TangS, et al., 2011. Changes in cortical bone response to high-fat diet from adolescence to adulthood in mice. Osteoporos Int, 22(8):2283-2293.

[32]KamineA, ShimozuruM, ShibataH, et al., 2012. Changes in blood glucose and insulin responses to intravenous glucose tolerance tests and blood biochemical values in adult female Japanese black bears (Ursus thibetanus japonicus). Jpn J Vet Res, 60(1):5-13.

[33]KatoM, PatelMS, LevasseurR, et al., 2002. Cbfa1-independent decrease in osteoblast proliferation, osteopenia, and persistent embryonic eye vascularization in mice deficient in Lrp5, a Wnt coreceptor. J Cell Biol, 157(2):303-314.

[34]KimJM, YangYS, ParkKH, et al., 2020. A RUNX2 stabilization pathway mediates physiologic and pathologic bone formation. Nat Commun, 11:2289.

[35]KimMS, YangYM, SonA, et al., 2010. RANKL-mediated reactive oxygen species pathway that induces long lasting Ca²⁺ oscillations essential for osteoclastogenesis. J Biol Chem, 285(10):6913-6921.

[36]KochFP, MerkelC, Al-NawasB, et al., 2011. Zoledronate, ibandronate and clodronate enhance osteoblast differentiation in a dose dependent manner—a quantitative in vitro gene expression analysis of Dlx5, Runx2, OCN, MSX1 and MSX2. J Cranio-Maxillofac Surg, 39(8):562-569.

[37]KomoriT, 2005. Regulation of skeletal development by the Runx family of transcription factors. J Cell Biochem, 95(3):445-453.

[38]KomoriT, 2011. Signaling networks in RUNX2-dependent bone development. J Cell Biochem, 112(3):750-755.

[39]KoshiharaY, HoshiK, 1997. Vitamin K₂ enhances osteocalcin accumulation in the extracellular matrix of human osteoblasts in vitro. J Bone Miner Res, 12(3):431-438.

[40]KurganN, BottKN, HelmecziWE, et al., 2019. Low dose lithium supplementation activates Wnt/β-catenin signalling and increases bone OPG/RANKL ratio in mice. Biochem Biophys Res Commun, 511(2):394-397.

[41]LamAP, GottardiCJ, 2011. β-Catenin signaling: a novel mediator of fibrosis and potential therapeutic target. Curr Opin Rheumatol, 23(6):562-567.

[42]LampropoulosCE, PapaioannouI, D'CruzDP, 2012. Osteoporosis—a risk factor for cardiovascular disease? Nat Rev Rheumatol, 8(10):587-598.

[43]LanaspaMA, KuwabaraM, Andres-HernandoA, et al., 2018. High salt intake causes leptin resistance and obesity in mice by stimulating endogenous fructose production and metabolism. Proc Natl Acad Sci USA, 115(12):3138-3143.

[44]LeeNK, SowaH, HinoiE, et al., 2007. Endocrine regulation of energy metabolism by the skeleton. Cell, 130(3):456-469.

[45]LeonardMB, ShultsJ, WilsonBA, et al., 2004. Obesity during childhood and adolescence augments bone mass and bone dimensions. Am J Clin Nutr, 80(2):514-523.

[46]LiuCT, BroeKE, ZhouYH, et al., 2017. Visceral adipose tissue is associated with bone microarchitecture in the framingham osteoporosis study. J Bone Miner Res, 32(1):143-150.

[47]LiuXL, LiangYZ, XiaN, et al., 2021. Decrease in leptin mediates rat bone metabolism impairments during high-fat diet-induced catch-up growth by modulating the OPG/RANKL balance. 3 Biotech, 11:103.

[48]MartinSA, SouderDC, MillerKN, et al., 2018. GSK3β regulates brain energy metabolism. Cell Rep, 23(7):‍1922-1931.e4.

[49]MinematsuA, NishiiY, SakataS, 2018. High-fat/high-sucrose diet results in higher bone mass in aged rats. Bone Rep, 8:18-24.

[50]MoorerMC, RiddleRC, 2018. Regulation of osteoblast metabolism by Wnt signaling. Endocrinol Metab, 33(3):318-330.

[51]NovinecM, LenarčičB, 2013. Cathepsin K: a unique collagenolytic cysteine peptidase. Biol Chem, 394(9):‍1163-1179.

[52]NúñezNP, CarpenterCL, PerkinsSN, et al., 2007. Extreme obesity reduces bone mineral density: complementary evidence from mice and women. Obesity, 15(8):1980-1987.

[53]PenroseHM, HellerS, CableC, et al., 2017. High-fat diet induced leptin and Wnt expression: RNA-sequencing and pathway analysis of mouse colonic tissue and tumors. Carcinogenesis, 38(3):302-311.

[54]PetitMA, BeckTJ, ShultsJ, et al., 2005. Proximal femur bone geometry is appropriately adapted to lean mass in overweight children and adolescents. Bone, 36(3):568-576.

[55]PickeAK, SylowL, MøllerLLV, et al., 2018. Differential effects of high-fat diet and exercise training on bone and energy metabolism. Bone, 116:120-134.

[56]PinedaN, OwenM, TuckerC, et al., 2017. Hibernating little pocket mice show few seasonal changes in bone properties. Anat Rec, 300(12):2175-2183.

[57]Pitynski-MillerD, RossM, SchmillM, et al., 2017. A high salt diet inhibits obesity and delays puberty in the female rat. Int J Obes, 41(11):1685-1692.

[58]PollockNK, LaingEM, HamrickMW, et al., 2011. Bone and fat relationships in postadolescent black females: a pQCT study. Osteoporos Int, 22(2):655-665.

[59]ProiettoJ, 2020. Obesity and bone. F1000Research, 9(Faculty Rev):1111.

[60]ReinholdMI, NaskiMC, 2007. Direct interactions of Runx2 and canonical Wnt signaling induce FGF18. J Biol Chem, 282(6):3653-3663.

[61]RiganoKS, GehringJL, HutzenbilerBDE, et al., 2017. Life in the fat lane: seasonal regulation of insulin sensitivity, food intake, and adipose biology in brown bears. J Comp Physiol B, 187(4):649-676.

[62]SavvidisC, TournisS, DedeAD, 2018. Obesity and bone metabolism. Hormones, 17(2):205-217.

[63]ShapsesSA, SukumarD, 2012. Bone metabolism in obesity and weight loss. Annu Rev Nutr, 32:287-309.

[64]TakagiY, SugimotoT, KobayashiM, et al., 2018. High-salt intake ameliorates hyperglycemia and insulin resistance in WBN/Kob-Lepr^fa/fa rats: a new model of type 2 diabetes mellitus. J Diabetes Res, 2018:3671892.

[65]TakahashiN, MaedaK, IshiharaA, et al., 2011. Regulatory mechanism of osteoclastogenesis by RANKL and Wnt signals. Front Biosci (Landmark Ed), 16(1):21-30.

[66]TauerJT, Boraschi-DiazI, al RifaiO, et al., 2021. Male but not female mice with severe osteogenesis imperfecta are partially protected from high-fat diet-induced obesity. Mol Genet Metab, 133(2):211-221.

[67]VijayGV, ZhaoN, den HollanderP, et al., 2019. GSK3β regulates epithelial-mesenchymal transition and cancer stem cell properties in triple-negative breast cancer. Breast Cancer Res, 21:37.

[68]VimalrajS, 2020. Alkaline phosphatase: structure, expression and its function in bone mineralization. Gene, 754:144855.

[69]WangHW, ChenFL, LiJX, et al., 2020. Vaspin antagonizes high fat-induced bone loss in rats and promotes osteoblastic differentiation in primary rat osteoblasts through Smad-Runx2 signaling pathway. Nutr Metab (Lond), 17:9.

[70]WosjeKS, KhouryPR, ClaytorRP, et al., 2009. Adiposity and TV viewing are related to less bone accrual in young children. J Pediatr, 154(1):79-85.e72.

[71]YanL, GraefGL, NielsenFH, et al., 2015. Soy protein is beneficial but high-fat diet and voluntary running are detrimental to bone structure in mice. Nutr Res, 35(6):‍523-531.

[72]YanX, NiuQH, GaoXL, et al., 2021. Differential protein metabolism and regeneration in gastrocnemius muscles in high-fat diet fed mice and pre-hibernation daurian ground squirrels: a comparison between pathological and healthy obesity. Zool Stud, 60:e6.

[73]YanagiharaGR, ShimanoRC, TidaJA, et al., 2017. Influence of high-fat diet on bone tissue: an experimental study in growing rats. J Nutr Health Aging, 21(10):1337-1343.

[74]YangY, MallampatiS, SunBH, et al., 2013. Wnt pathway contributes to the protection by bone marrow stromal cells of acute lymphoblastic leukemia cells and is a potential therapeutic target. Cancer Lett, 333(1):9-17.

[75]YunTJ, TallquistMD, AicherA, et al., 2001. Osteoprotegerin, a crucial regulator of bone metabolism, also regulates B cell development and function. J Immunol, 166(3):‍1482-1491.

[76]ZhongL, YuanJH, HuangL, et al., 2020. RANKL is involved in Runx2-triggered hepatic infiltration of macrophages in mice with NAFLD induced by a high-fat diet. Biomed Res Int, 2020:6953421.

[77]ZhouS, WangGH, QiaoL, et al., 2018. Age-dependent variations of cancellous bone in response to ovariectomy in C57BL/6J mice. Exp Ther Med, 15(4):3623-3632.