慢性氟中毒对大鼠神经细胞线粒体融合基因表达及线粒体形态的影响  

Changed expression of mito-fusion 1 and mitochondrial fragmentation in the cortical neurons of rats with chronic fluorosis

作　　者：楼迪栋[1,2] 潘际刚[1] 张凯琳[1] 秦双立[1] 刘燕斐[1] 于燕妮[1] 官志忠[1,3] 

机构地区：[1]贵阳医学院病理学教研室,550000 [2]贵阳医学院法医司法鉴定中心 [3]贵州省分子生物学重点实验室

出　　处：《中华预防医学杂志》2013年第2期170-174,共5页Chinese Journal of Preventive Medicine

基　　金：基金项目：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[黔科合LG字（2011）007号,黔省专合字（2012）162号]

摘　　要：目的研究慢性氟中毒对大鼠神经细胞线粒体融合基因Mfnl表达及线粒体形态的影响。方法将60只1月龄清洁级SD大鼠以单纯随机法分为3组，每组20只，雌雄各半，分笼喂养。以饮水中加入氟剂量的不同，将大鼠分为对照组、低氟组、高氟组，染毒剂量分别为0、10和50mg／L，分别在饲养3、6个月后，观察氟斑牙程度。采用透射扫描电子显微镜观察线粒体形态。用western—blotting和免疫组织化学方法检测线粒体融合基因Mfnl表达。结果染氟组大鼠出现不同程度的氟斑牙，3个月时低氟组以I度氟斑牙为主（16只），高氟组发生I度、Ⅱ度氟斑牙分别为11、9只；6个月时低氟组发生I度、Ⅱ度、Ⅲ度氟斑牙分别为14、6、0只，高氟组为6、13、1只；对照组均未出现。低氟组、高氟组在3个月时尿氟分别为（3．30±1．18）、（5．10±0．35）mg／L（F=3．18，P〈0．05）；6个月时分别为（4．16±1．39）、（5．70±1．70）mg／L（F=3．17，P〈0．05）。3个月时，对照组大鼠大脑皮质神经细胞线粒体形态变化不明显，染氟组大鼠神经细胞线粒体均出现球状空泡状改变。Mfnl的蛋白免疫组织化学强阳性表达神经细胞数发生了改变，与对照组[（53．0±4．54）个]相比较，低氟组[（51．09±6．25）个]的改变无统计学意义（t=1．7，／9〉0．05），而高氟组[（59．71±5．64）个]增加，差异有统计学意义（t=2．1，P〈0．05）；weste．Flq-blotting法测得蛋白印迹平均吸光度值也具有相同的改变，与对照组（1．21±0．18）相比，低氟组（1．22±0．26）的改变无统计学意义（t=1．1，P〉0．05），高氟组（1．66±0．20）的水平升高，差异有统计学意义（t=2．1，P〈0．05）。6个月时低氟组、高氟组大鼠大脑皮质神经细胞线粒体出现嵴消失、球状空泡状改变，对照组无明显改变。Mfnl的蛋白免疫组织化学强阳性�Objective To observe the mitochondrial fragmentation and the expression of mito- fusion 1 gene in the cortical neurons of rats with chronic fluorosis, and to reveal their roles in mitochondria damage to neurons due to chronic fluorosis. Methods SD rats were divided randomly into three groups of 20 each （ a half females and a half males housed individually in stainless-steel cages）, and fed with the different doses of fluoride containing in drinking water （ untreated control containing 0 mg/L fluoride, and low-fluoride and high supplemented with 10 and 50 mg/L fuoride, respectively ）. After 3 or 6 months exposure,the mitochondrial morphology of the neurons in rat brains were observed by transmission electron microscopy （TEM）, then the expression of mitoehondrial fusion gene, Mfnl, were detected by immunohistochemistry and western-blotting, respectively. Results Dental fluorosis was obvious in the rats exposed to excessive fluoride in their drinking water, that is, （ 16 rats out of 20 ） numbers of I o detal fluorosis in the low-flouride group,and （ 11 rats out of 20） numbers of I ° and （9 rats out of 20） numbers of II ° detal fluorosis in the high-flouride group were observed after 3 months exposure. Moreover, （ 14 rats out of 20） numbers of I ° and （ 6 rats out of 20） numbers of II ° detal fluorosis in the low-flouride group and （ 6 rats out of 20） numbers of Io, （ 13 rats out of 20） numbers of II °, and （ 1 rats out of 20） numbers of III ° detal fluorosis in the high-flouride group were observed after 6 months exposure. And both of untreated controls without detal fluorosis were also observed. The urinary, level of fluoride in the low-flouride group （3.30 ± 1.18） mg/L and in the high-fluoride group （5.10 ±0. 35） were observed after 3 months exposure （F = 3.18, P 〈 0. 05 ）. Moreover, the urinary level of fluoride in the low-flouride group （4. 16 ± 1.39） mg/L and in the high-fluoride group （5.70 ± 1.70） mg/L were also observed after 6 month

关 键 词：氟中毒 牙 大鼠 线粒体融合基因Mfn1 神经系统 

分 类 号：R135[医药卫生—劳动卫生]