摘要目的 探讨高氟对成骨细胞骨钙素(OC)及小鼠糖脂代谢的影响.方法 常规培养3T3-L1前脂肪细胞和MC3T3-E1成骨细胞,0、2、10、50 mg/L氟化钠(NaF)刺激MC3T3-E1成骨细胞建立染氟细胞模型,检测OC水平,以OC水平升高最显著所对应的NaF浓度为最佳染氟条件.细胞实验分为4组:3T3-L1、3T3-L1+NaF、3T3-L1+MC3T3-E1、3T3-L1+MC3T3-E1+NaF组,其中处理组分别或联合加以最佳条件的NaF(50mg/L)和MC3T3-E1成骨细胞共培养,采用酶联免疫吸附试验(ELISA)检测细胞OC和脂联素(APN)表达水平.同时,将40只C57BL/6小鼠按体质量从小到大进行编号,遵照随机数字表法分为对照组和染氟组,每组20只,对照组饮用纯净水,染氟组饮用含100 mg/L的NaF溶液,观察小鼠牙齿和体质量[M(P25,P75)]变化.12周后采用ELISA法检测血清中OC和APN水平,葡萄糖氧化酶法检测空腹血清葡萄糖(FPG)水平,化学发光法检测空腹胰岛素(FINS)水平,并采用胰岛素抵抗指数稳态模型(HOMA-IR)评估胰岛素抵抗情况.结果 3T3-L1、3T3-L1+NaF、3T3-L1+MC3T3-E1、3T3-L1+MC3T3-E1+NaF组APN表达水平分别为(0.94±0.18)、(1.07±0.21)、(1.76±0.20)、(2.49±0.43)μg/L,MC3T3-E1细胞与NaF对APN表达水平有协同促进效应(F=14.519,P＜ 0.01).小鼠染氟后12周体质量[27.5 (25.8,28.3)g]显著低于同期对照小鼠[31.4(30.3,32.6)g,Z=-4.695,P＜ 0.01].染氟组小鼠FPG[(7.78±1.86)mmol/L]、FINS[(3.22±0.75) mU/L]、OC[(6.11±1.49)μg/L]、APN[(8.65±1.78)μg/L]及HOMA-IR (1.15±0.49)均高于对照组[(5.40±1.51) mmol/L、(2.45±0.64)mU/L、(3.14±0.92)μg/L、(4.03±1.45) μg/L、0.62±0.31,t=-4.466、-3.518、-7.560、-9.002、-4.182,P均＜0.01].小鼠OC水平与FPG、FINS、APN、HOMA-IR呈正相关(r=0.868、0.707、0.911、0.818,P均＜0.01).结论 小鼠成骨细胞OC在高氟作用下显著升高,OC与血糖、APN水平密切相关,是高氟影响糖脂代谢的关键分子之一.

abstractsObjective To explore excessive fluoride on expression of osteocalcin (OC) in osteoblast and glucolipid metabolism in mice.Methods 3T3-L1 preadipocyte and MC3T3-E1 osteoblast were conventionally cultured,MC3T3-E1 cells were stimulated with O,2,10 and 50 mg/L sodium fluoride (NaF) to establish fluorine cell model,and levels of OC were tested.The corresponding NaF concentration resulted in the highest OC level was used as the optimal fluorine concentration.Cell experiments were divided into four groups:3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF.The treatment groups were respectively or jointly treated with the optimal concentration of NaF (50 mg/L) or MC3T3-E1 osteoblast,enzyme linked immunosorbent assay (ELISA) was used to detect OC and adiponectin (APN) levels.At the same time,40 C57BIL/6 mice were numbered by weight,randomly divided into control and fluoride groups,20 per group,control group drunk pure water,fluoride group drunk 100 mg/L NaF solution,changes of teeth and body weight [M (P25,P75)] of the mice were observed.Serum OC and APN levels were tested by ELISA at 12 weeks after modeling.The fasting plasma glucose (FPG) and the fasting plasma insulin (FINS) were detected by glucose oxidase and chemiluminescence methods,and insulin resistance (HOMA-IR) was evaluated.Results The APN levels of 3T3-L1,3T3-L1 + NaF,3T3-L1 + MC3T3-E1 and 3T3-L1 + MC3T3-E1 + NaF groups were (0.94 ± 0.18),(1.07 ± 0.21),(1.76 ± 0.20),and (2.49 ± 0.43) μg/L,MC3T3-E1 osteoblast with NaF had promote collaborative interaction effect on APN levels (F =14.519,P ＜ 0.01).The body weight of mice in fluoride group [27.5 (25.8,28.3) g] was significantly lower than that of the control group [31.4 (30.3,32.6) g,Z =-4.695,P ＜ 0.01].The levels of FPG [(7.78 ± 1.86) mmol/L],FINS [(3.22 ± 0.75) mU/L],OC [(6.11 ± 1.49) μμg/L],APN [(8.65 ± 1.78) μg/L] and HOMA-IR (1.15 ± 0.49) were higher than those of control groups [(5.40 ± 1.51) mmol/L,(2.45 ± 0.64) mU/L,(3.14 ± 0.92) μg/L,(4.03 ± 1.45) μg/L,0.62 ± 0.31],the differences were statistically significant (t =-4.466,-3.518,-7.560,-9.002,-4.182,P ＜ 0.01).OC levels in mice were positively correlated with FPG,FINS,APN and H-OMA-IR (r =0.868,0.707,0.911,0.818,P ＜ 0.01).Conclusion The OC of osteoblast in mice exposed to fluoride is increased significantly,OC levels in mice are closely related to blood glucose and APN,it is one of the key molecules in lipid metabolism.