耳缘静脉移植自体骨髓间充质干细胞治疗家兔氟骨症效果观察  

Effects of transplantation of autologous bone marrow mesenchymal stem cells via ear vein injection on rabbits with skeletal fluorosis

作　　者：李晨 蒋菲菲 雷平贵 邹迅 陈义家 潘雪莉 LI Chen;JIANG Fei-fei;LEI Ping-gui;ZOU Xun;CHEN Yi-jia;PAN Xue-li(Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Department of Toxicology, School of Public Health, Guizhou Medical University, Guiy~mg, Guizhou 550025, Chin)

机构地区：[1]贵州医科大学公共卫生学院贵州医科大学环境污染与疾病监控教育部重点实验室,贵州贵阳550025 [2]贵州医科大学附属医院医学影像科

出　　处：《环境与健康杂志》2018年第2期114-117,F0003,共5页Journal of Environment and Health

基　　金：贵州省优秀青年科技人才培养对象专项资金[黔科合人字(2013)07号]

摘　　要：目的观察耳缘静脉移植自体骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)对家兔氟骨症治疗的有效性和可行性。方法取家兔54只,雌雄各半,随机分为空白对照组(12只)和氟骨症模型组(42只),分别饮用含氟化钠0、300 mg/L的自来水。90 d后将42只模型组家兔随机分为示踪组(单次/多次耳缘静脉注射、示踪溶剂对照组,6只家兔/组)和治疗组(单次/多次耳缘静脉注射、自然恢复组、治疗溶剂对照组,6只家兔/组),雌雄各半。分离、培养、鉴定治疗组氟骨症家兔BMSCs,分别以单次及多次耳缘静脉移植回氟中毒家兔体内。示踪组中,用5-溴脱氧尿嘧啶核苷(Brd U)标记BMSCs移植入家兔体内,免疫组化检测示踪BMSCs。治疗组中,60 d后,观察治疗前后骨骼X线影像学变化,以定量CT(QCT)测定家兔活体股骨骨密度,以酶联免疫吸附法(ELISA)测定家兔血清碱性磷酸酶(ALP)及骨钙素(BGP),以氟离子选择电极法测定家兔股骨骨氟含量,以HE染色观察股骨组织病理学改变并计算骨小梁面积。结果免疫组化结果显示多次耳缘静脉移植示踪组呈Brd U阳性表达,成骨细胞胞核呈淡黄色,而单次耳缘静脉移植示踪组、示踪溶剂对照组、示踪空白对照组、自然恢复组未见Brd U表达。多次耳缘静脉移植组治疗后X线影像显示股骨斑片状高密度影消失,紊乱骨小梁有所恢复;骨密度[(562.42±15.69)g/cm3]、ALP活力[(24.88±3.82)U/L]、BGP浓度[(1247.12±291.52)μg/L]、骨氟含量[(1699.16±528.69)mg/kg]、骨小梁面积[(28.71±1.88)Tb.Ar]均较自然恢复组下降[(635.11±93.42)g/cm3、(43.08±2.82)U/L、(3207.73±788.80)μg/L、(2557.65±173.90)g/kg、(38.52±2.81)Tb.Ar,P均<0.05],差异具有统计学意义;多次耳缘静脉移植组HE染色显示骨髓腔间隙增大,骨小梁数目减少,同时伴有部分新生成的粗细不均、排列整齐、骨髓间隙较均匀的正常骨小梁。而单次耳缘静脉移植组BMSCs未观察到�Objective To observe the efficacy and feasibility of transplantation of autologous bone marrow mesenchymal stem cells via ear vein injection on rabbits with skeletal fluorosis. Methods A total of 54 rabbits, half male and half female, were randomly divided into control group（n =12） and experimental model group（n =42）. The two groups of rabbits were given drinking water containing Na F at 0 or 300 mg/L respectively. After 90-day treatment, 42 rabbits were divided into tracer groups and treatment groups. The tracing groups included single tracer group, multiple tracer group and tracer solvent control group. The treatment groups included single treatment group, multiple treatment group, spontaneous recovery group and treatment solvent control group（six rabbits in each group, half male and half female）. After isolation, culture and identification of the bone marrow mesenchymal stem cells（BMSCs） from rabbits with skeletal fluorosis, autologous BMSCs were transplanted into rabbit bodies via intravenous at once or three times, respectively. In tracer groups, BMSCs were isolated from rabbits with skeletal fluorosis and labeled with Brd U, then the cells were implanted into rabbits by ear vein injection. BMSCs were detected by immunohistochemistry. Femur image was observed by X-ray,femur bone mineral density was examined with QCT, the alkaline phosphatase（ALP） and osteocalcin（BGP） in serum were measured using ELISA,the bone fluoride content was determined by the fluorine ion-selective electrode method,histopathology changes of femur were observed by HE staining and the trabecular acreage was calculated 60 days after transplantation. Results Immunohistochemical results showed that Brd U was observed in the multiple tracer groups, the nucleus of osteoblasts was yellowish. But Brd U was not observed in the single tracer groups, tracer solvent control group, and tracer blank control group. In the multiple treatment group, X-ray showed patchy high-density images of femur were disappeared and abnormal bo

关 键 词：氟骨症 家兔 骨髓间充质干细胞 移植 

分 类 号：R994.6[医药卫生—毒理学]