摘要
背景:目前利用人羊膜间充质干细胞促进骨细胞分化成熟的相关研究缺乏,采用移植人羊膜间充质干细胞治疗骨质疏松很可能为骨质疏松的治疗打开新的视野。目的:探讨人羊膜间充质干细胞移植对去卵巢骨质疏松大鼠骨代谢指标和骨组织微结构的影响及机制。方法:选择雌性未孕的Wistar大鼠36只,随机分组,模型组23只行双侧卵巢切除,假手术组13只不切除卵巢。术后3个月,各组均取3只处死,ELISA法测定血清抗酒石酸酸性磷酸酶和骨碱性磷酸酶水平,苏木精-伊红染色观察股骨结构的病理改变,判断造模情况。造模成功后,将模型组剩余20只大鼠随机分为2组,对照组和治疗组分别通过尾静脉注射氯化钠注射液和人羊膜间充质干细胞,2次/周,连续6周。细胞移植6周后,通过ELISA法测定血清抗酒石酸酸性磷酸酶和骨碱性磷酸酶水平,RT-PCR、Western blot法检测大鼠股骨转录因子Runx2、Osterix基因和蛋白表达,苏木精-伊红染色观察骨组织微结构变化。结果与结论:(1)造模3个月后,模型组大鼠血清骨碱性磷酸酶水平明显下降,抗酒石酸酸性磷酸酶水平明显升高;苏木精-伊红染色结果显示,股骨骨小梁稀疏、数目明显减少甚至断裂,而假手术组骨小梁结构较完整,证明造模成功;(2)人羊膜间充质干细胞移植后6周,与对照组相比,治疗组骨碱性磷酸酶水平明显升高,抗酒石酸酸性磷酸酶水平明显下降(P<0.05);治疗组大鼠股骨转录因子Runx2、Osterix表达明显增加(P<0.05);(3)细胞移植后6周,治疗组大鼠股骨新生骨小梁广泛形成,骨小梁结构破坏较对照组明显减轻;(4)结果表明,人羊膜间充质干细胞移植对去卵巢骨质疏松具有治疗作用,可能是通过促进Runx2、Osterix表达发挥作用。
BACKGROUND： There are rare studies directly exploring the use of human amniotic mesenchymal stem cells to promote differentiation and maturation of osteocytes. Therefore, human amniotic mesenchymal stem cell transplantation is likely to open new horizons for the treatment of osteoporosiss. OBJECTIVE： To explore the effect of human amniotic mesenchymal stem cell transplantation on bone metabolism index and bone microstructure in ovariectomized osteoporosis rats and the relevant mechanisms. METHODS： Thirty-six female Wistar rats without pregnancy were randomly divided into sham and model group. Twenty-three rats in the model group were ovariectomized, and thirteen rats in the sham group received no treatment. Three months after ovariectomy, three rats from each group were executed. ELISA was used to detect serum levels of tartrate-resistant acid phosphatase （TRACP） and bone alkaline phosphatase （BALP）. Hematoxylin-eosin staining was performed to observe the pathological changes of the bone. After successful modeling, the remaining rats in the model group were randomized into control and treatment groups, respectively given tail vein injection of sodium chloride and human amniotic mesenchymal stem cells, twice a week, for consecutive 6 weeks. Six weeks after cell transplantation, ELISA was used to detect serum levels of TRACP and BALP; RT-PCR and western blot assay were used to detect expressions of Runx2 and Osterix mRNA and protein and hematoxylin-eosin staining was used to observe bone microstructure changes. RESULTS AND CONCLUSION： At 3 months after ovariectomy, the rats in the model group exhibited significantly decreased BALP and remarkably increased TRACP as well as sparse number of bone trabeculae some of which were even ruptured shown by hematoxylin-eosin staining, However, the bone trabeculae of rats in the sham group still remained intact. These results indicated the successful modeling in the model group. At 6 weeks after cell transplantation, significantly increased BALP, Runx2, Osterix
出处
《中国组织工程研究》
CAS
北大核心
2017年第1期92-97,共6页
Chinese Journal of Tissue Engineering Research
