慢性氟中毒大鼠骨组织Gli1、β-catenin表达变化及其与骨形成增强间的相关性分析  

Change and relationship between Gli1 and β-catenin on rats′ bone formation with chronic fluorosis

作　　者：邓超男 张颖[1,3] 徐淋 赵丽娜 令狐艳 于燕妮[1,2] Deng Chaonan;Zhang Ying;Xu Lin;Zhao Lina;Linghu Yan;Yu Yanni(Department of Pathology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Pathology,Guizhou Medical University,Guiyang 550004,China;Maternal and Child Care Hospital of Guiyang,Guiyang 550004,China)

机构地区：[1]贵州医科大学附属医院病理科,贵阳550004 [2]贵州医科大学病理学教研室,贵阳550004 [3]贵阳市妇幼保健院,贵阳550004

出　　处：《中华病理学杂志》2020年第2期168-173,共6页Chinese Journal of Pathology

基　　金：国家自然科学基金(81260419);高等学校博士学科点专项科研基金博导类资助课题(20125215110001);贵州省自然科学基金(黔科合LH字[2017]7192号、黔科合基础[2018]1135号、黔科合平台人才[2018]5779-37);贵州省研究生科研基金(KYJJ2017010)。

摘　　要：目的观察使用刺猬蛋白(Hedgehog)信号阻断剂环巴胺(cyclopamine,Cycl)对慢性氟中毒大鼠骨组织中神经胶质瘤相关癌基因同源蛋白1(glioma-associated oncogene homolog 1,Gli1)及β链蛋白(β-catenin)表达的影响,探讨两者表达变化在氟致骨形成增强中的意义。方法SD大鼠48只分为4组,每组12只。对照组饮用自来水(含氟量<1 ppm),氟中毒组(F组)、氟+环巴胺组(F+Cycl组)和氟+二甲基亚砜组(F+DMSO组)分别饮用含50 ppm氟化钠(NaF)的自来水。饲养6个月后分别给F+Cycl组和F+DMSO组大鼠腹腔注射环巴胺和DMSO。检测各组大鼠尿氟含量;酶联免疫吸附试验测定血清骨碱性磷酸酶(bone alkaline phosphatase,BALP)含量;HE染色观察骨组织形态计量学变化;实时荧光定量PCR法、免疫组织化学和蛋白印迹法检测骨组织中Gli1和β-catenin的mRNA和蛋白表达情况。结果染氟后F组、F+Cycl组和F+DMSO组大鼠尿氟含量、骨小梁密度、骨小梁宽度较对照组明显增加,但3组间未见明显差异(P>0.05)。F组较对照组血清BALP含量增加,Cycl阻断后血清BALP含量明显降低(P<0.05)。与对照组比较,F组Gli1和β-catenin mRNA和蛋白均有表达增强。使用Cycl后,两者表达均较F组降低(P<0.05),而F+DMSO组则无明显变化。Gli1与β-catenin的表达呈正相关关系(r=0.476,P<0.05);Gli1、β-catenin蛋白表达与血清BALP、骨小梁密度分别呈正相关关系(r1=0.457,r2=0.466,r3=0.581,r4=0.554,P<0.05)。结论Cycl可阻断慢性氟中毒大鼠骨组织Gli1表达,并且其影响成骨相关因子β-catenin的表达,这可能参与到氟致骨形成增强的过程中。Objective To investigate the change and association of glioma-associated oncogene homolog 1(Gli1)andβ-catenin on bone formation in rats with chronic fluorosis which were inhibited by cyclopamine(Cycl).Methods 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(NaF<1 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 andβ-catenin in bone tissue were detected using real-time PCR,immunohistochemistry and Western blot.Results 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(P<0.05).The expression of Gli1 andβ-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 andβ-catenin(r=0.476,P<0.05).The expression of Gli1 andβ-catenin was also associated with BALP concentration and volume of bone trabeculae,respectively(r1=0.457,r2=0.466,r3=0.581,r4=0.554,respectively,P<0.05 for all).Conclusions 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 ofβ-catenin,which is an osteogenesis factor.

关 键 词：大鼠 骨和骨组织 氟中毒 

分 类 号：R5[医药卫生—内科学]