氟和铝在饮茶型氟中毒大鼠关节软骨细胞凋亡中的联合作用及其机制研究

Exploration on the combined role and the mecbanism of fluorine and aluminum on articular chondrocyte apoptosis in rats induced by drinking brick-tea fluorosis

目的 研究氟、铝在饮茶型氟中毒大鼠关节软骨细胞凋亡中的联合作用，探讨其作用机制。方法 4周龄雄性Wistar大鼠按体质量随机分成5组，即对照组、氟组、铝组、氟＋铝组、砖茶组。对照组饮自来水，氟组饮氟化钠水溶液，铝组饮氯化铝水溶液，氟＋铝组饮氟化钠和氯化铝混合的水溶液，砖茶组饮浓缩的砖茶水溶液，实验期3个月。TUNEL法检测关节软骨细胞凋亡的表达情况，RT-PCR和免疫组化技术检测关节软骨诱生型一氧化氮合酶（iNOS）和半胱氨酸蛋白酶蛋白-3（Caspase-3）mRNA及蛋白表达情况。结果 ①与对照组比较，各实验组软骨细胞凋亡指数均明显升高（P〈0．01），其中氟＋铝组软骨细胞凋亡指数最高（P〈0．05）。②氟、铝联合对iNOSm RNA相对表达升高具有协同作用（F=682．737，P〈0．01）。③氟、铝单独或同时摄人，均可上调关节软骨组织iNOS和Caspase-3在蛋白质水平上的表达。结论氟、铝协同促进饮茶型氟中毒大鼠关节软骨细胞凋亡的发生。

Objective To investigate the combined role of fluorine and aluminum on articular chondrocyte apoptosis in rats induced by drinking tea fluorosis, and to explore the mechanism. Methods 4-week male Wistar rats were randomized into 5 groups according to their body weights： control group, fluoride treated group, aluminum treated group, fluoride with aluminum treated group, and brick-tea treated group. The experiment lasted for 3 months. TUNEL was used to detect apoptosis expression of articular chondrocytes, and the mRNA and protein expression of iNOS, Caspase-3 in articular cartilage were determined by RT-PCR and immunohistochemistry techniques. Results ① Compared with the control group, the chondrocyte apoptosis index was higher in every experiment group （P 〈 0.01 ） and was the highest in the fluorine with treated group（P 〈 0.05）. ②The combination of fluorine with aluminum had a synergistic effect on the relative mRNA levels of iNOS（F = 682.737, P 〈 0.01 ）. ③ Fluorine and aluminum, solely or simultaneity ingested, could upregulate the protein expression of iNOS, Caspase-3 in articular cartilage. Conclusions To some degree, combination of fluorine with aluminum promotes chondrocyte apoptosis.