为了研究氟对小鼠附睾结构形态及紧密连接相关基因的影响,将60只健康雄性小鼠随机分成4组,对照组饮用蒸馏水,3个氟处理组分别饮用含氟化钠(NaF)25、50和100mg/L的蒸馏水。染氟60d,取小鼠股骨,采用氟离子选择电极法测其氟含量;取右侧附睾制作石蜡切片,HE染色后进行组织形态结构观察;左侧附睾提取RNA,采用实时荧光定量PCR技术(qRT-PCR)检测紧密连接相关基因的表达。结果显示,与对照组相比,氟处理组小鼠股骨氟含量随摄氟剂量的增加呈剂量依赖性,其中50、100mg/L剂量组差异极显著(P〈0.01);氟处理组附睾尾管腔厚度极显著增厚(P〈0.01);紧密连接相关基因Claudin-2 mRNA表达量极显著下降(P〈0.01),100mg/L剂量组ZO-1mRNA表达量显著下降(P〈0.05)。摄入过量NaF影响附睾形态及其紧密连接相关基因的表达,从而可能对血-附睾屏障造成损伤。
The purpose of this study was to examine the effects of fluoride on epididymal morphological structure and mRNA expression of tight junction genes. 60 healthy male mice were divided into four groups randomly, including control group (distilled water) and 25,50,100 mg NaF/L groups, respectively. After 60 days, the thighbones were collected and fluoride contents were determined by fluorine ion selective electrode method. Paraffin slides of right epididymis with HE stain were produced to evaluate epididymal morphological structure under light microscope. The left epididymis were collected to isolate total RNA and detect the mRNA expression of tight junction genes by qRT-PCR. The results showed that the fluoride contents of thighbones increased in a dose-dependent manner, and were extremely significantly higher in 50 and 100 mg/L groups than those in control group (P〈0.01). NaF extremely significantly increased the thickness of cauda epididymal pipe wall in fluoride groups (P〈0. 01) and the Claudin-2 mRNA expressions in fluoride groups were extremely significantly decreased (P〈0.01) and the ZO-1 mRNA expression in 100 mg/L group was significantly decreased (P〈0.05) as compared with control group. In conclusion, excessive NaF ingestion adversely affected epididymal morphological structure and mRNA expressions of Claudin-2 and ZO-1 ,which might contribute to the damage of blood-epididymal barrier.