摘要
目的研究慢性氟中毒大鼠大脑皮质神经细胞线粒体分裂基因动态相关蛋白1（Drp1）表达的改变。方法将120只sD大鼠（1月龄、体重100—120g）以单纯随机法分为3组（对照组、低氟组和高氟组），每组40只，各组染毒剂量以饮水中加入氟剂量计算：分别为0、10和50mS／L；染毒3个月和6个月时，每组分别处死20只。采用免疫组织化学方法和免疫蛋白印迹法（western—blotting）检测线粒体分裂蛋白Drp1表达。结果3个月时，Drpl的蛋白免疫组织化学强阳性表达神经细胞个数发生了改变，与对照组[（36．3±5．8）个]相比较，低氟组[（34．7±4．1）个]的改变无统计学意义（t=1．5，P〉0．05），而高氟组[（45．0±d．7）个]表达增加（t=8．8，P〈0．05）；western—blotting法蛋白印迹平均灰度值也具有相同的改变，与对照组（0．59-t-O．03）相比，低氟组（0．62±0．03）和高氟组（0．71±0．02）的水平升高（t值分别为0．02、0．11，P值均〈0．05）。6个月时，Drpl的蛋白免疫组织化学强阳性表达神经细胞个数发生了改变，与对照组[（33．2±4．4）个]相比较，低氟组[（36．6±3．8）个]和高氟组[（39．4±4．2）个]升高（t值分别为3．5，6．3，P值均〈0．05）；western—blotting法蛋白印迹平均值也有改变，与对照组（0．65-t-O．06）相比，低氟组（0．804-0．09）和高氟组（0．764-0．08）的水平升高（t值分别为0．1、0．1，P值均〈0．05）。结论高剂量染氟可造成分裂基因Drp1表达改变，慢性氟中毒神经细胞损伤可能与线粒体分裂亢进有关。
Objective To explore the changes of protein expression of mito-fission gene dynamin- related 1 （ Drp 1 ） in the cortical neurons of rats with chronic fluarosis. Methods A total of 120 one-month- old SD rats （each weighing approximately 100 - 120 g at the beginning of the experiment） were randomly divided into three groups, and fed with the different doses of fluoride containing in drinking water （ untreated control containing 0 mg/L fluoride, and low-fluoride ＆ high-fluoride supplemented with 10 and 50 mg/L fluoride, respectively）. After 3 or 6 months exposure, 20 rats from each group were killed. Then the protein expression of mitochondrial fission gene, Drpl, was detected by immunohistochemistry and western-blotting method. Results Dental fluorosis and urinary fluorosis were obviously found in the rats exposed to fluoride. At the experiment period of 3 months, the numbers of positive cells of Drpl detected by immunohistochemistry changed. Compared with the control group （36. 3 ± 5.8 ）, the changes in low-fluoride group （ 34. 7 ± 4. 1 ） showed no significant difference （ t = 1.5, P 〉 0. 05 ）, but the increase in high-fluoride group （45.0 ± 4. 7 ） had statistical significance （ t = 8.8, P 〈 0. 05 ）. The western-blotting method had consistent results. Compared with the control group （ 0. 59 ± 0. 03 ）, a significant increase of the average topical density in low- fluoride （0.62 ±_ O. 03 ） and high-fluoride （0.71 ± O. 02） groups were found （ t =0. 02,0. 11 ,P 〈 0.05）. At the experiment period of 6 months ,the numbers of positive cells of Drpl detected by immunohistochemistry significantly changed. Compared with the control group （ 33.2 ± 4. 4 ） , the number in low-fluoride and high-fluoride groups were separately （ 36. 6 ± 3.8 ） and （ 39.4 ± 4. 2 ） , both increased significantly （ t =3.5,6. 3, P 〈 0. 05 ）. Same results could be found in western-blotting method, compared with the control group （0. 65 ±0. 06） ,the average topical density in low- fluoride （0. 80 ±0. 09） and high- fluoride （0. 76 ± 0. 08） groups both increased significantly （ t = 0. 1,0. 1 ,P 〈 0. 05 ）. Conclusions Taking excessive amount of fluoride might result in the changes of expression of Drpl, and the neurons damage from the chronic fluorosis might be associated with the hyperfunction of mitochondrial fusion.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2013年第6期561-564,共4页
Chinese Journal of Preventive Medicine
基金
科技部国际合作项目（2010DFB30530）
国家自然科学基金（81160335）
贵州省科技基金项目{黔科合LG字[2011]007号、黔省专合字（2012）162号}
温州市科技局（Y20080099）
