【作者】 刚亚栋；

【导师】 张本忠； 王俊玲；

【作者基本信息】 兰州大学 ， 卫生毒理学， 2010， 硕士

【摘要】 为了探讨氟中毒对肾脏的毒性作用及其可能机制,为氟病的研究提供理论依据。本研究选择健康Wistar大鼠32只,雌雄各半,体重150-180g,随机分为4组,每组8只,即：生理盐水组(对照组)、低氟组(100mg·kg-1·d-1 NaF)中氟组(200mg·kg-1·d-1 NaF)、高氟组(300mg·kg-1·d-1 NaF),连续灌胃染毒90天,每三天称体重一次。染毒结束次日,摘眼球采血；颈椎脱臼处死大鼠,迅速分离肾脏,分别称质量并计算肾脏脏器系数；制作肾脏组织HE染色切片,观察大鼠肾组织病理形态学的改变；用生化分析仪检测肾功能生化指标：血清尿素、肌酐；运用原子吸收光谱仪检测大鼠血清铜、锌、钙、镁、铁5种微量元素的含量；取单侧肾脏组织用HNO3-KCIO4低温湿法消化,利用空气-乙炔火焰原子吸收光谱法测定大鼠肾脏组织中铜、锌、钙、镁、铁5种微量元素的含量；制备肾脏组织匀浆,分光光度法检测肾脏组织过氧化氢酶(CAT)活性、丙二醛(MDA)和过氧化氢(H2O2)的含量。研究结果如下：1、染毒第30天各组大鼠体重,组间差异有统计学意义(P<0.05),其中低氟组、中氟组、高氟组与对照组比较差异无统计学意义(P>0.05)。低氟组、中氟组与高氟组比较差异有统计学意义(P<0.05)；第0、60、90天各组间比较差异无统计学意义,其中低氟组、中氟组、高氟组与对照组比较差异无统计学意义(P>0.05)。2、灌胃90天后,肾脏脏器系数,各染氟组与对照组比较组间比较差异无统计学意义(P>0.05)。3、光镜观察,高氟组肾脏近端小管轻度水肿,近曲小管上皮细胞水肿,伴有不同程度颗粒变性,部分呈空泡变性,间质小血管轻度扩张。4、与对照组比较,高氟组血清Urea含量有显著性升高(P<0.01)。同时与低氟组比较,中、高氟组血清Urea含量亦有显著性升高(P<0.05)。血清肌肝(Cr)含量随染毒剂量的增加有升高趋势,但与对照组比较无显著性差异(P>0.05)。5、与对照组比较,低氟组、高氟组Cu含量和中氟组、高氟组Fe含量降低(P<0.05)。与低氟组比较,中氟组Fe含量降低(P<0.05)。与中氟组比较,高氟组Cu含量降低(P<0.05)。6、与对照组比较,中氟组的锌含量、高氟组的铁含量和低氟组的钙含量的有显著性降低(P<0.05)。与低氟组比较,中、高氟组Fe含量有显著性较低(P<0.01)；中、高氟组Ca含量有显著性升高(P<0.05)。与中氟组比较,高氟组Zn含量有显著性升高(P<0.05)。7、与对照组比较,高氟组的CAT活性有显著性降低,但MDA有显著性升高(P<0.05)。与低氟组比较,高氟组的CAT活性有显著性降低(P<0.05)；与中氟组比较,高氟组的MDA有显著性升高(P<0.05)。本研究结果显示,氟对肾脏组织具有一定的损害作用。其机制可能与氟降低肾脏组织中金属元素的含量和改变肾脏的组织结构及引起肾脏组织氧化应激损伤有关,其具体机制有待进一步的研究。更多还原

【Abstract】 To probe into the fluoride-induced renal toxicity and its possible mechanisms in rats so as to provide theoretical basis for the prevention and treatment of fluorosis. A total of 32 with male and female evenly weighting 150～180g were randomly divided into 4 groups, control group with normal saline, the low fluoride group of 100 mg·kg-1·d-1 NaF, the medium fluoride group of 200 mg·kg-1·d-1 NaF and the high fluoride group of 300 mg·kg-1·d-1 NaF, with 8 rats in each group. The rats were exposed to fluoride by intragastric administration for 90 days. The weight of the rats was measured one time every 3 days. The day after the last intragastric administration, the blood samples were collected on eyes of rats and all rats were killed. The Rat kidneys were promptly plucked off and the organ coefficient of kidney was calculated. The tissues of kidney were made into pathological sections and the pathological changes of rat kidney were observed by light microscopy. Some indexes of renal function such as serum urea, serum creatinine were measured with biochemical analyzers. The levels of the five trace elements in serum of rats were measured with Atomic Absorption Spectrophotometer. One of two kidneys from each Rat was digested with HNO3-KClO4 at low temperature. Five trace elements in kidney tissue were determined by air-acetylene flame atomic absorption spectrometry, and another one kidney were prepared as homogenate to detect the content of malondialdehyde (MDA), Hydrogen peroxide (H2O2) and the activity of catalase (CAT).The results indicated that:1.There were statistic differences among groups in the weight of rats on exposure 30 days. Compared with control group, the weight of rats among three groups showed no significant difference (P>0.05). However, the weight of rats in low and medium fluoride groups after 30 days exposure was significant increased compared with high fluoride group (P<0.05), but significant differences were not observed between after 60 days and 90 days exposure.2.The organ coefficient of kidney among four groups showed no significant difference (P>0.05).3.Under the light microscopy, granular degeneration to some extent was observed in the epithelial cells of kidney proximal tubule, some cells showed vacuolar degeneration, and also mildly congestion and swelling in mesenchymal blood vessels were observed.4.The serum urea of the high fluoride group showed significant increase compared with control group (P<0.01). Meanwhile, the serum urea of the medium group and high fluoride group showed significant increase compared with the low fluoride group (P<0.05). The serum creatinine showed no difference between any each of fluoride exposure groups compared with the control group (P>0.05).5.The Cu2+ contents in rat serum of the low group and high fluoride groups showed significant decrease compared with control group (P<0.05). The Fe3+ contents in rat serum of the medium and high fluoride group showed significant decrease compared with control group (P<0.01). Meanwhile, The Fe3+ contents in rat serum of the medium fluoride group showed significant decrease compared with control group (P<0.05). Compared with the medium fluoride group, the Cu2+ contents in rat serum of high fluoride groups showed significant decrease(P<0.05). The contents of Ca2+, Mg2+ and Zn2+ in rat serum were not different in each group compared with the control group (all P>0.05).6.The Zn2+ contents in kidney tissue from the medium fluoride group showed significant decrease compared with control group (P<0.05). The Fe3+ contents in kidney of the high fluoride groups showed significant decrease compared with control group (P<0.05). The Ca2+ contents in kidney tissue of the low fluoride groups showed significant decrease compared with control group (P<0.05). Meanwhile, the Fe3+ contents in kidney of the medium and high fluoride groups showed significant decrease compared with low fluoride group (P<0.01). The Ca2+ contents in kidney tissue of the medium and high fluoride groups showed significant increase compared with low fluoride group (P<0.05). Compared with the medium fluoride group, The Zn2+ contents in kidney tissue from the medium fluoride group showed significant increase compared with medium group (P<0.05). The contents of Mg2+ and Cu2+ in kidney were not different in each group compared with the control group (all P>0.05).7.Compared with the control group, the activity of CAT in kidney tissue from the high fluoride group showed significant decrease (P<0.05). The content of MDA showed significant increase compared with the control group (P<0.05). Compared with the low group, the activity of CAT in kidney tissue from the high fluoride group showed significant decrease (P<0.05). The content of MDA showed significant increase compared with the medium group (P<0.05). The contents of H2O2 in kidney were not different in each group compared with the control group (all P>0.05).The results indicated that fluoride could cause damage to the kidney of rat. Fluoride could reduce the content of metallic elements, and cause significant change in renal tissue and lead to some disorder in redox equilibrium of kidney tissue, which might be related to its damage mechanism. However, its specific mechanism needed to further study.更多还原