Deng C N, Zhang Y, Xu L, Zhao L N, Linghu Y, Yu Y N
Department of Pathology, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China; Department of Pathology, Guizhou Medical University, Guiyang 550004, China.
Department of Pathology, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China; Maternal and Child Care Hospital of Guiyang, Guiyang 550004, China.
Zhonghua Bing Li Xue Za Zhi. 2020 Feb 8;49(2):168-173. doi: 10.3760/cma.j.issn.0529-5807.2020.02.012.
To investigate the change and association of glioma-associated oncogene homolog 1 (Gli1) and β-catenin on bone formation in rats with chronic fluorosis which were inhibited by cyclopamine (Cycl). Forty-eight Sprague-Dawley rats were evenly divided to four groups, including control, F, F+Cycl and F+DMSO groups. The control group were fed with tap water (NaF1 ppm). The F, F+Cycl and F+DMSO groups were exposed to NaF (50 ppm) in drinking water as the chronic fluorosis model. Then the rats in F+Cycl or F+DMSO groups were injected by Cycl or DMSO after 6 months, respectively. Urine fluoride concentration was detected using fluorine ion selective electrode. The enzyme-linked immunosorbent assay (ELISA) was used to detect bone alkaline phosphatase (BALP). Bone tissues were stained with hematoxylin-eosin. The mRNA and protein expression of Gli1 and β-catenin in bone tissue were detected using real-time PCR, immunohistochemistry and Western blot. Compared with the controls, the urine fluoride concentration and the width and volume of bone trabeculae were increased in the F, F+Cycl and F+DMSO groups, but no statistical difference among the 3 fluorosis groups. The concentration of BALP was increased in the F group and decreased in F+Cycl group (0.05). The expression of Gli1 and β-catenin mRNA and protein was higher in the F and F+Cycl groups than controls, but lower in the F+Cycl group than in the F group. There was positive correlation between the expression of Gli1 and β-catenin (0.476, 0.05). The expression of Gli1 and β-catenin was also associated with BALP concentration and volume of bone trabeculae, respectively (0.457, (2)0.466, 0.581, 0.554, respectively, 0.05 for all). The expression of Gli1 can be inhibited by Cycl. It may be involved in the bone formation of rats with chronic fluorosis. It may also affect the expression of β-catenin, which is an osteogenesis factor.
为研究环杷明（Cycl）抑制慢性氟中毒大鼠中胶质瘤相关癌基因同源物1（Gli1）和β-连环蛋白的变化及其与骨形成的关系。将48只Sprague-Dawley大鼠平均分为四组，即对照组、F组、F+Cycl组和F+DMSO组。对照组饮用自来水（含1 ppm氟化钠）。F组、F+Cycl组和F+DMSO组饮用含50 ppm氟化钠的水作为慢性氟中毒模型。6个月后，分别对F+Cycl组和F+DMSO组大鼠注射Cycl或二甲基亚砜（DMSO）。采用氟离子选择性电极检测尿氟浓度。用酶联免疫吸附测定法（ELISA）检测骨碱性磷酸酶（BALP）。骨组织进行苏木精-伊红染色。采用实时荧光定量聚合酶链反应（PCR）、免疫组织化学和蛋白质免疫印迹法检测骨组织中Gli1和β-连环蛋白的mRNA和蛋白表达。与对照组相比，F组、F+Cycl组和F+DMSO组尿氟浓度及骨小梁宽度和体积增加，但3个氟中毒组之间无统计学差异。F组BALP浓度升高，F+Cycl组降低（P<0.05）。F组和F+Cycl组Gli1和β-连环蛋白mRNA及蛋白表达高于对照组，但F+Cycl组低于F组。Gli1与β-连环蛋白表达呈正相关（r=0.476，P<0.05）。Gli1和β-连环蛋白表达分别与BALP浓度和骨小梁体积相关（r分别为0.457、0.466、（2）0.581、0.554，P均<0.05）。Cycl可抑制Gli1表达。它可能参与慢性氟中毒大鼠的骨形成。它也可能影响作为成骨因子的β-连环蛋白的表达。
