不同剂量氟对大鼠骨组织Ihh、Shh、Smo mRNA和蛋白表达的影响  

Expressions of Ihh,Shh and Smo mRNA and protein in rats'bone exposed to different doses of fluoride and the significance

作　　者：邓超男 张颖[2,3] 徐淋[2,3] 赵丽娜 令狐艳[2] 于燕妮 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,550004

出　　处：《中华地方病学杂志》2020年第9期630-635,共6页Chinese Journal of Endemiology

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);贵州省科技合作计划项目(黔科合LH字[2017]7192号);贵州省自然科学基金(黔科合基础[2018]1135号);贵州医科大学2018年度学术新苗培养及创新探索专项项目(黔科合平台人才[2018年]5779-37);贵州省研究生科研基金(KYJJ2017010);贵阳医学院博士启动基金(院博合J字[2014]013号)。

摘　　要：目的探讨刺猬蛋白(Hedgehog,Hh)信号通路相关因子(Ihh、Shh、Smo)在慢性氟中毒大鼠骨组织中的表达变化及意义。方法36只健康SD大鼠,根据体质量(100~120 g)采用随机数字表法分为3组(12只/组,雌雄各半);对照组大鼠饮用自来水(含氟量<1 mg/L),低氟组和高氟组染氟剂量分别为5、50 mg/L。各组大鼠饲养6个月时,收集24 h尿样,取大鼠股骨干骺端,氟离子选择电极法检测尿氟、骨氟含量;HE染色,光镜观察骨组织形态学改变;酶联免疫吸附法检测血清骨碱性磷酸酶(BALP)水平;实时荧光定量PCR(Real-time PCR)和免疫组织化学法检测骨组织Ihh、Shh、Smo mRNA和蛋白表达水平。结果随着染氟含量的增加,各组大鼠尿氟、骨氟含量和血清BALP水平[尿氟:(1.37±0.44)、(5.96±0.56)、(7.60±0.61)mg/L,骨氟:(306.04±12.58)、(652.91±51.83)、(1094.11±126.34)mg/kg,血清BALP:(27.78±4.09)、(46.59±5.75)、(57.45±3.99)U/L]逐渐增加(P均<0.05)。光镜下染氟组大鼠股骨干骺端呈骨质硬化表现。高氟组大鼠Ihh、Shh、Smo mRNA表达水平(1.39±0.36、0.56±0.23、0.40±0.15)高于对照组和低氟组(0.73±0.19、0.92±0.34,0.19±0.04、0.36±0.16,0.14±0.04、0.24±0.13,P均<0.05),低氟组Shh mRNA表达水平高于对照组(P<0.05)。高氟组大鼠Ihh、Smo蛋白表达水平(138.89±3.72、149.29±7.63)高于对照组和低氟组(127.39±2.69、134.81±3.53,129.64±12.62、139.07±9.30),且低氟组高于对照组(P均<0.05);高氟组大鼠Shh蛋白表达水平(141.26±7.49)高于对照组(130.96±11.10,P<0.05)。结论Hh信号通路相关因子在氟中毒大鼠骨组织中出现表达改变,氟可能通过诱导Hh信号活化来影响骨形成。Objective To investigate the expression changes of Hedgehog related factors(Ihh,Shh and Smo)in bone of rats with chronic fluorosis,and the significance.Methods Thirty-six healthy SD rats were divided to three groups with the method of random digits table by body weight(100-120 g),12 rats in each group,half male and half female.The rats of control were fed with tap water(NaF<1 mg/L),and the experimental rats were exposed to NaF(low dose fluoride group:5 mg/L,high dose fluoride group:50 mg/L)added to the drinking water to establish the chronic fluorosis model.After the rats were raised for six months,24-hour urine samples were collected and the femoral metaphysis of the rats was taken.Urine fluoride and bone fluoride were detected by fluorin ion selective electrode method.Bone tissues were stained with hematoxylin-eosin and observed under light microscope.The content of bone alkalinephosphatase(BALP)in rats'serum was detected by enzyme-linked immunosorbent assay(ELISA).The expressions of Ihh,Shh and Smo mRNA and protein in bone were detected by Real-time PCR and immunohistochemistry(IHC).Results The contents of urine fluoride,bone fluoride and serum BALP were increased gradually in the control,low and high doses fluoride groups[urine fluoride:(1.37±0.44),(5.96±0.56),(7.60±0.61)mg/L;bone fluoride:(306.04±12.58),(652.91±51.83),(1094.11±126.34)mg/kg;BALP:(27.78±4.09),(46.59±5.75),(57.45±3.99)U/L,P<0.05].It could observed that bone sclerosis by light microscope in low and high doses fluoride groups.The expressions of Ihh,Shh and Smo mRNA in high dose fluoride group(1.39±0.36,0.56±0.23,0.40±0.15)were higher than those of the control and low dose fluoride groups(0.73±0.19,0.92±0.34;0.19±0.04,0.36±0.16;0.14±0.04,0.24±0.13;P<0.05).The expression of Shh mRNA in low dose fluoride group was higher than that of the control group(P<0.05).The expressions of Ihh and Smo protein in high dose fluoride group(138.89±3.72,149.29±7.63)were higher than those of the control and the low dose fluoride groups(127.39±2.6

关 键 词：骨和骨组织 氟中毒 刺猬蛋白 

分 类 号：R599.1[医药卫生—内科学]