高盐饮食通过激活破骨细胞促进小鼠骨质疏松发生
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摘要
目的明确高盐饮食对小鼠破骨细胞分化的影响及其促进骨质疏松的作用。方法获得C57小鼠骨髓单个核细胞,在体外定向诱导破骨分化体系中比较对照组(未加入氯化钠)、低剂量组(20mmol/L氯化钠)及高剂量组(40 mmol/L氯化钠)破骨细胞生成数量;将24只C57小鼠随机平均分为正常组(普通饲料喂养)、卵巢切除组(切除卵巢,普通饲料喂养)、高盐饮食组(切除卵巢,8%高盐饲料+生理盐水喂养),6周后分析3组左侧股骨破骨细胞生成数量并测定血清骨吸收及骨保护指标水平;选择右侧股骨进行显微CT扫描,比较3组骨组织形态计量学参数。结果体外高盐状态破骨细胞生成数量明显增加;体内高盐饮食组破骨细胞生成数量明显高于其它两组,I型胶原羧基端肽和核因子κB受体活化因子配体不同程度增加,骨保护素水平显著降低,骨体积分数、骨皮质厚度、骨小梁数量和骨小梁厚度下降尤为明显,骨表面积/骨体积、骨小梁分离度和骨小梁模式因子不同程度增高。结论高盐饮食可通过激活小鼠破骨细胞分化及其功能活化增强骨吸收作用,进而诱导骨质疏松发生。
Objective To clarify the promotion effect of mouse osteoclast differentiation and osteoporosis by high-salt dietary. Methods Bone marrow mononuclear cells of C57 mice were obtained and induced to differentiate into osteoclasts. During this procedure, cells were cultured in each of 3 kinds of medium: no salt addition(control group), addition of 20 mmol/L salt(low-dose group), and addition of 40 mmol/L salt(high-dose group). The quantity of formed osteoclasts was acquired and compared among all 3 groups. In in-vivo study, a total of 24 C57 mice were divided into 3 groups randomly and equally: a normal group(regular feeding), an ovariectomy group(removal of ovary and regular feeding), and a high-salt group(removal of ovary, fed with 8% salt and drinking normal saline). At 6 weeks after administration, in all 3 cohorts, the number of generated osteoclasts in left femurs, as well as indicators associated with bone resorption and protection in serum were acquired. Moreover, bone histomorphometric parameters in right femurs were obtained and compared between each group using micro-CT. Results High-salt medium ex vitro enhanced the formation of osteoclasts significantly. Similarly, in vivo study found the largest number of osteoclasts in the high-salt group in comparison with the other 2 cohorts. Also in this group, C-telopeptide of type I collagen and receptor activator of nuclear factor-κ B ligand increased at different levels, while osteoprotegerin decreased remarkably. With respect to bone histomorphometric parameters in the high-salt group, bone volume/total volume, cortical wall thickness, trabecular number and trabecular thickness were significantly increased whereas bone surface area/bone volume, trabecular spacing and trabecular pattern factor were increased at various levels. Conclusions High-salt dietary enables the occurrence of osteoporosis through activating differentiation of bone marrow mononuclear cells into osteoclasts and strengthening of overall bone-resorption potential.
Objective To clarify the promotion effect of mouse osteoclast differentiation and osteoporosis by high-salt dietary. Methods Bone marrow mononuclear cells of C57 mice were obtained and induced to differentiate into osteoclasts. During this procedure, cells were cultured in each of 3 kinds of medium: no salt addition(control group), addition of 20 mmol/L salt(low-dose group), and addition of 40 mmol/L salt(high-dose group). The quantity of formed osteoclasts was acquired and compared among all 3 groups. In in-vivo study, a total of 24 C57 mice were divided into 3 groups randomly and equally: a normal group(regular feeding), an ovariectomy group(removal of ovary and regular feeding), and a high-salt group(removal of ovary, fed with 8% salt and drinking normal saline). At 6 weeks after administration, in all 3 cohorts, the number of generated osteoclasts in left femurs, as well as indicators associated with bone resorption and protection in serum were acquired. Moreover, bone histomorphometric parameters in right femurs were obtained and compared between each group using micro-CT. Results High-salt medium ex vitro enhanced the formation of osteoclasts significantly. Similarly, in vivo study found the largest number of osteoclasts in the high-salt group in comparison with the other 2 cohorts. Also in this group, C-telopeptide of type I collagen and receptor activator of nuclear factor-κ B ligand increased at different levels, while osteoprotegerin decreased remarkably. With respect to bone histomorphometric parameters in the high-salt group, bone volume/total volume, cortical wall thickness, trabecular number and trabecular thickness were significantly increased whereas bone surface area/bone volume, trabecular spacing and trabecular pattern factor were increased at various levels. Conclusions High-salt dietary enables the occurrence of osteoporosis through activating differentiation of bone marrow mononuclear cells into osteoclasts and strengthening of overall bone-resorption potential.
引文
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[16]Frick KK,LaPlante K,Bushinsky DA.RANK ligand and TNF-alpha mediate acid-induced bone calcium efflux in vitro[J].Am J Physiol Renal Physiol,2005,289(5):F1005-F1011.
[17]Krieger NS,Frick KK,Bushinsky DA.Mechanism of acid-induced bone resorption[J].Curr Opin Nephrol Hypertens,2004,13(4):423-436.
[18]Albano G,Dolder S,Siegrist M,et al.Increased bone resorption by osteoclast-specific deletion of the sodium/calcium exchanger isoform 1(NCX1)[J].Pflugers Arch,2017,469(2):225-233.
[2]Baccaro LF,Conde DM,Costa-Paiva L,et al.The epidemiology and management of postmenopausal osteoporosis:a viewpoint from Brazil[J].Clin Interv Aging,2015,10:583-591.
[3]Tella SH,Gallagher JC.Prevention and treatment of postmenopausal osteoporosis[J].J Steroid Biochem Mol Biol,2014,142:155-170.
[4]Park SM,Joung JY,Cho YY,et al.Effect of high dietary sodium on bone turnover markers and urinary calcium excretion in Korean postmenopausal women with low bone mass[J].Eur J Clin Nutr,2015,69(3):361-366.
[5]Wu L,Luthringer BJC,Feyerabend F,et al.Increased levels of sodium chloride directly increase osteoclastic differentiation and resorption in mice and men[J].Osteoporos Int,2017,28(11):3215-3228.
[6]Iwasa Y,Shimoyama K,Aoki K,et al.The effect of high salt intake on the mandibular bone loss in Dahl-Iwai salt-sensitive rat[J].J Med Dent Sci,2000,47(3):187-195.
[7]Hernandez AL,Kitz A,Wu C,et al.Sodium chloride inhibits the suppressive function of FOXP3+regulatory T cells[J].J Clin Invest,2015,125(11):4212-4222.
[8]Cho JH,Zhang R,Kilfoil PJ,et al.Delayed repolarization underlies ventricular arrhythmias in rats with heart failure and preserved ejection fraction[J].Circulation,2017,136(21):2037-2050.
[9]Teucher B,Dainty JR,Spinks CA,et al.Sodium and bone health:impact of moderately high and low salt intakes on calcium metabolism in postmenopausal women[J].J Bone Miner Res,2008,23(9):1477-1485.
[10]Frings-Meuthen P,Buehlmeier J,Baecker N,et al.High sodium chloride intake exacerbates immobilization-induced bone resorption and protein losses[J].J Appl Physiol(1985),2011,111(2):537-542.
[11]Creedon A,Cashman KD.The effect of high salt and high protein intake on calcium metabolism,bone composition and bone resorption in the rat[J].Br J Nutr,2000,84(1):49-56.
[12]Buehlmeier J,Remer T,Frings-Meuthen P,et al.Glucocorticoid activity and metabolism with NaCl-induced low-grade metabolic acidosis and oral alkalization:results of two randomized controlled trials[J].Endocrine,2016,52(1):139-147.
[13]Buehlmeier J,Frings-Meuthen P,Remer T,et al.Alkaline salts to counteract bone resorption and protein wasting induced by high salt intake:results of a randomized controlled trial[J].J Clin Endocrinol Metab,2012,97(12):4789-4797.
[14]Robey IF.Examining the relationship between diet-induced acidosis and cancer[J].Nutr Metab(Lond),2012,9(1):72-82.
[15]Frings-Meuthen P,Baecker N,Heer M.Low-grade metabolic acidosis may be the cause of sodium chloride-induced exaggerated bone resorption[J].J Bone Miner Res,2008,23(4):517-524.
[16]Frick KK,LaPlante K,Bushinsky DA.RANK ligand and TNF-alpha mediate acid-induced bone calcium efflux in vitro[J].Am J Physiol Renal Physiol,2005,289(5):F1005-F1011.
[17]Krieger NS,Frick KK,Bushinsky DA.Mechanism of acid-induced bone resorption[J].Curr Opin Nephrol Hypertens,2004,13(4):423-436.
[18]Albano G,Dolder S,Siegrist M,et al.Increased bone resorption by osteoclast-specific deletion of the sodium/calcium exchanger isoform 1(NCX1)[J].Pflugers Arch,2017,469(2):225-233.