代佑罡, 陈宣好, 杨璐珲, 张华, 韦艳. 氟化钠饮水染毒6个月对大鼠骨组织成骨活性标志的影响[J]. 环境与职业医学, 2019, 36(8): 725-730. DOI: 10.13213/j.cnki.jeom.2019.19123
引用本文: 代佑罡, 陈宣好, 杨璐珲, 张华, 韦艳. 氟化钠饮水染毒6个月对大鼠骨组织成骨活性标志的影响[J]. 环境与职业医学, 2019, 36(8): 725-730. DOI: 10.13213/j.cnki.jeom.2019.19123
DAI You-gang, CHEN Xuan-hao, YANG Lu-hui, ZHANG Hua, WEI Yan. Effects of six-month sodium fluoride exposure via drinking water on osteogenic activity markers in rat bone tissues[J]. Journal of Environmental and Occupational Medicine, 2019, 36(8): 725-730. DOI: 10.13213/j.cnki.jeom.2019.19123
Citation: DAI You-gang, CHEN Xuan-hao, YANG Lu-hui, ZHANG Hua, WEI Yan. Effects of six-month sodium fluoride exposure via drinking water on osteogenic activity markers in rat bone tissues[J]. Journal of Environmental and Occupational Medicine, 2019, 36(8): 725-730. DOI: 10.13213/j.cnki.jeom.2019.19123

氟化钠饮水染毒6个月对大鼠骨组织成骨活性标志的影响

Effects of six-month sodium fluoride exposure via drinking water on osteogenic activity markers in rat bone tissues

  • 摘要: 背景 氟中毒是由于长期从空气、食物和饮水中摄入过量的氟而引起的以氟骨症和氟斑牙为主要特征的一种慢性全身性疾病。地方性氟中毒在全国分布广泛且严重,受威胁人口众多。研究氟中毒相关机制有助于预防和治疗因摄氟过多而造成的氟骨症、氟斑牙等骨相病变。

    目的 探讨不同浓度氟化钠染毒对大鼠成骨活性标志的影响,为研究氟中毒机制提供实验数据。

    方法 将32只SD大鼠按体重均衡随机均分为:对照组(0 mg/L)、低氟组(50 mg/L)、中氟组(150 mg/L)、高氟组(250 mg/L)。采取饮水染毒方式,染氟至6个月时分别收集各组大鼠12 h尿液,测定尿氟浓度;大鼠经10%水合氯醛麻醉,心脏采血处死后,测定血氟浓度;取左前肢,液氮研磨,采用实时荧光PCR法测定Col ⅠRunx2Osx mRNA表达水平;取右前肢,液氮研磨,采用Western blot检测Col Ⅰ、Runx2和Osx的蛋白表达水平。

    结果 染毒期间,大鼠饮食、饮水正常,精神状况良好,不同染毒组与对照组相比体重差异均无统计学意义。染氟6个月后,低、中、高染氟组大鼠尿氟含量(9.71±2.33)、(20.36±4.92)、(42.36±4.56)mg/kg均高于对照组(5.69±1.59)mg/kg,差异有统计学意义(P < 0.05);低、中、高染氟组大鼠血氟含量(0.19±0.01)、(0.23±0.01)、(0.29±0.02)mg/L均高于对照组(0.14±0.01)mg/L,差异有统计学意义(P < 0.05);且随着染氟剂量的增加,尿氟和血氟都呈上升趋势(分别r=0.89、0.90,均P < 0.01)。染氟6个月后,低、中、高染氟组大鼠Col Ⅰ mRNA表达分别为(0.91±0.28)、(0.87±0.33)、(0.67±0.31)均低于对照组(1.31±0.43),低、中、高染氟组大鼠Runx2 mRNA表达分别为(0.81±0.33)、(0.78±0.18)、(0.57±0.25)均低于对照组(1.18±0.28),差异均有统计学意义(P < 0.05);且随着染氟剂量的增加,Col ⅠRunx2 mRNA的表达都呈下降趋势(r=-0.56、-0.31,均P < 0.05)。与对照组(1.01±0.15)相比,低、中染氟组大鼠Osx mRNA表达量分别为(1.31±0.20)、中氟组(1.49±0.41)升高,但高氟组(0.58±0.15)降低,差异均有统计学意义(P < 0.05)。染氟6个月后,低、中、高染氟组大鼠Col Ⅰ蛋白相对表达量分别为(0.17±0.02)、(0.15±0.01)、(0.11±0.03)均低于对照组(0.22±0.03),差异有统计学意义(P < 0.05);且随着染氟剂量的增加,Col Ⅰ蛋白的相对表达量呈下降趋势(r=-0.88,P < 0.05)。与对照组分别为(0.19±0.03)、(0.16±0.03)相比,低氟组分别为(0.30±0.04)和(0.25±0.02)、中氟组(0.26±0.01)和(0.31±0.03)大鼠的Runx2蛋白和Osx蛋白的相对表达量升高,高氟组(0.12±0.02)和(0.10±0.03)降低,差异均有统计学意义(P < 0.05)。

    结论 氟对骨组织成骨活性标志表达的影响复杂,Col Ⅰ的表达随着染毒剂量的增加呈下降趋势;而中、低剂量染毒会促进Runx2和Osx的表达,高剂量时则会抑制其表达。

     

    Abstract: Background Fluorosis is a chronic systemic disease characterized by skeletal fluorosis and dental fluorosis caused by long-term intake of excessive fluorine from air, food, and water. Endemic fluorosis is widespread throughout China, and severely threatens a large populaton. To study the mechanism of fluorosis is helpful to prevent and treat skeletal fluorosis, dental fluorosis, and other bone diseases caused by excessive intake of fluoride.

    Objectve This experiment is designed to investgate the effects of different doses of sodium fluoride on osteogenic actvity markers in rats, and to provide experimental data for studying the mechanism of fluorosis.

    Methods Thirty-two SD rats were randomly divided into one control group (0 mg/L) and three fluoride groups (50, 150, and 250 mg/L, respectvely). Afer six months of fluoride administraton via drinking water, urinary samples were collected within 12 h to detect urinary fluoride concentration; then the rats were executed after 10% chloral hydrate anesthesia, and cardiac blood samples were collected to detect serum fluoride concentraton; lef forelimbs were ground in liquid nitrogen, and the mRNA expressions of Col Ⅰ, Runx2, and Osx were detected by real-time quanttatve PCR; right forelimbs were ground in liquid nitrogen, and the protein expressions of Col Ⅰ, Runx2, and Osx were detected by Western blot.

    Results During the fluoride administration period, the rats were in good mental status with normal dietary intake and water consumpton, and there was no signifcant difference in body weight between the exposure groups and the control group. Afer the designed six-month treatment, the urinary fluoride concentratons of the fluoride groups(9.71±2.33), (20.36±4.92), and (42.36±4.56) mg/kg were statstcally higher than that of the control group(5.69±1.59) mg/kg (P < 0.05), so were the serum fluoride concentratons(0.19±0.01), (0.23±0.01), and (0.29±0.02) mg/L vs. (0.14±0.01) mg/L (P < 0.05); as the fluoride exposure dose increased, the urinary and serum fluoride concentratons showed rising trends (r=0.89, 0.90, Ps < 0.01). Afer the treatment for six months, the mRNA expression levels of Col Ⅰ in the lowdose group (0.91±0.28), the medium-dose group (0.87±0.33), and the high-dose group (0.67±0.31) were statstcally higher than that in the control group (1.31±0.43) (P < 0.05); the mRNA expression levels of Runx2 in the low-dose group (0.81±0.33), the medium-dose group (0.78±0.18), and the high-dose group (0.57±0.25) were statstcally higher than that in the control group (1.18±0.28) (P < 0.05); as the fluoride exposure dose increased, the mRNA expression levels of Col Ⅰ and Runx2 showed decreasing trends (r=-0.56, -0.31, Ps < 0.05). Compared with the control group (1.01±0.15), the mRNA expression levels of Osx in the low-dose group (1.31±0.20) and the medium-dose group (1.49±0.41) were increased, and that in the high-dose group (0.58±0.15) was decreased (P < 0.05). Afer the treatment for six months, the protein expression levels of Col Ⅰ in the low-dose group (0.17±0.02), the medium-dose group (0.15±0.01), and the high-dose group (0.11±0.03) were statstcally lower than that in the control group (0.22±0.03) (P < 0.05); as the fluoride exposure dose increased, the protein expression level of Col Ⅰ showed a downward trend (r=-0.88, P < 0.05). Compared with the control group(0.19±0.03), (0.16±0.03), the protein expression levels of Runx2 and Osx in the low-dose group(0.30±0.04), (0.25±0.02) and the medium-dose group(0.26±0.01), (0.31±0.03) were increased, but the expression levels in the highdose group(0.12±0.02), (0.10±0.03) were decreased (P < 0.05).

    Conclusion The effect of fluoride on the expressions of osteogenic actvity markers is complex. As the fluoride exposure dose increases, the Col Ⅰ protein expression level shows a downward trend; whereas, the Runx2 and Osx protein expression levels of the low- and medium-dose fluoride groups are up-regulated, and those of the high-dose fluoride group are down-regulated.

     

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