摘要
目的:探讨Netrin-1增强骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)治疗骨质疏松性骨折的作用及机制。方法:流式细胞术检测BMSC凋亡水平和人白细胞抗原G(human leukocyte antigen-G,HLA-G)的表达;构建骨质疏松性骨折大鼠模型;ELISA检测碱性磷酸酶、抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)和骨钙素的浓度;免疫组化检测骨组织中人来源的BMSC和巨噬细胞的数目。结果:流式细胞术检测显示Netrin-1能够抑制缺血缺氧引起的BMSC凋亡(F=27.311,P=0.000),同时升高缺血缺氧抑制的HLA-G表达(F=27.094,P=0.000)。BMSC可减少血清中碱性磷酸酶和TRAP的浓度,升高血清中骨钙素浓度、骨组织内骨细胞数目和骨密度。Netrin-1能够进一步增强BMSC的这些功能,相对于BMSC组,差异均具有统计学意义。免疫组化结果显示,BMSC组存活的BMSC数目为(10.401±1.392)个/高倍视野(high-power field,HPF),Netrin-1组的数目为(25.506±2.257)个/HPF,2组之间差异有统计学意义(t=5.694,P=0.000);BMSC组巨噬细胞数目为(21.900±4.458)个/HPF,相对于骨质疏松组,差异有统计学意义;Netrin-1组巨噬细胞数目为(11.500±3.808)个/HPF,相对于BMSC组,差异具有统计学意义(F=79.863,P=0.000)。结论:Netrin-1通过细胞保护和调控炎症反应,增强BMSC对骨质疏松性骨折的治疗效果。
Objective:To investigate the role of Netrin-1-enhanced bone marrow mesenchymal stem cells(BMSC)on the treatment of osteoporotic fracture and its mechanism. Methods:The apoptotic level of BMSC and the expression of human leukocyte antigen-G(HLA-G)were detected by flow cytometry. Mice models with osteoporotic fracture were built. Concentrations of alkaline phosphatase,tartrate resistant acid phosphatase(TRAP)and osteocalcin were detected by ELISA,and the number of human BMSC and macrophages in bone tissue was detected by immunohistochemistry. Results:Flow cytometry showed that Netrin-1 was able to inhibit the apoptosis of BMSC induced by ischemia and hypoxia(F=27.311,P=0.000),and increase the expression of HLA-G(F=27.094,P=0.000). BMSC was able to reduce the concentration of alkaline phosphatase and TRAP in serum,and increase the concentration of osteocalcin in serum,the number of osteocytes in bone tissue and the bone mineral density. Netrin-1 was able to further enhance functions of BMSC.Compared with the BMSC group,the difference was statistically significant. Immunohistochemical results showed that the number of surviving BMSCs in the BMSC group was(10.401±1.392)/high-power field(HPF)and was(25.506±2.257)/HPF in the Netrin-1 group,with significant differences between two groups(t =5.694,P =0.000). The number of macrophages in the BMSC group was(21.900±4.458)/HPF,which was significantly different from that in the osteoporosis group. The number of macrophages in the Netrin-1 group was(11.500±3.808)/HPF,which was significantly different from that in the BMSC group(F=79.863,P=0.000). Conclusion:Netrin-1 can enhance the therapeutic effect of BMSC on osteoporotic fracture through cell protection and regulation of inflammatory reaction.
作者
王滋润
梁丽芹
肖成伟
郝鹏
Wang Zirun;Liang Liqin;Xiao Chengwei;Hao Peng(Department of Orthopedics,Sichuan Academy of Medical Sciences/Sichuan Provincial People's Hospital;Surgical Department,Sichuan Academy of Medical Sciences/Sichuan Provincial People's Hospital)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2021年第3期284-288,共5页
Journal of Chongqing Medical University
