摘要
目的探讨瘦素对绝经后骨质疏松病人骨髓间质干细胞(BMSCs)增殖分化的影响以及相关机制研究。方法留取20例因股骨颈骨折行股骨头置换股骨头,分离获得骨髓间质干细胞,利用成骨诱导培养液诱导培养,检测BMSCs诱导前后细胞表面抗原表达,将细胞分为5组:A组、B组、C组和D组分别加入10、1×10~2、1×10~3、1×10~4μg·L^(-1)浓度的瘦素(leptin),对照组不加入leptin。MTT法检测各组成骨样细胞增殖能力,实时荧光定量PCR检测各组细胞中ALP、OCN、RANKL和OPN基因表达。结果相比于诱导前,诱导后细胞表面CD44和HLA-DR表达量显著增加,而CD90和CD105表达量下降(P<0.05);与24 h时相比,A组、B组、C组、D组和对照组48、72、96 h时细胞增殖能力均增强(P<0.05);与对照组相比,A组、B组和C组48、72、96 h时细胞增殖能力显著增强(P<0.05),尤其是B组促增殖能力最强,而D组各时点细胞增殖能力则降低(P<0.05);与24 h时相比,A组、B组、C组、D组和对照组48、72、96 h时细胞中ALP、OCN和OPN mRNA相对表达量和OPN/RANKL比值均升高(P<0.05),且呈现随时间推移逐渐升高趋势,而RANKL mRNA相对表达量均降低(P<0.05),且呈现随时间推移逐渐降低趋势;A组、B组、C组和D组组间相比,48、72、96 h时细胞中ALP、OCN和OPN mRNA相对表达量和OPN/RANKL比值:B组>C组>A组>D组,而RANKL mRNA相对表达量:D组>A组>C组>B组。结论 leptin可促进绝经后骨质疏松病人BMSCs定向分化后成骨样细胞增殖分化,且具有时间依赖性,但存在最宜浓度。
Objective To investigate the effect of Leptin on the proliferation and differentiation of bone marrow mesenchymal stem cells （BMSCs） in postmenopausal osteoporosis patients and its related mechanism. Methods 20 cases of femoral heads were collected from patients with femoral neck fracture undergoing femoral head replacement. Bone marrow mesenchymal stem cells （ BMSCs） were isolated. The osteogenic induction medium was used to induce the culture. The cells were divided into five groups：group A,group B,group C and group D were given 10,1 x 102,1 x 103 and 1 x 104 jjig ？ L 1 leptin respectively,while the control group was not given leptin. The pro-liferation of osteoblast-like cells were detected by MTT assay. The expressions of ALP ,0CN,RANKL and OPN genes were detected by real-time fluorescent quantitative PCR. Results Compared with before induction, the expression levels of CD44 and HLA-DR in cells after induction were significantly increased （ P 〈 0. 05 ） , while the expression levels of CD90 and CD105 were decreased （ P 〈 0. 05 ）. Compared with the 24 h,the cell proliferation ability at 48,72 h and 96h in the group A,group B,group C,group D and control group were increased （ P 〈 0. 05 ）. Compared with the control group, the proliferation abilities at 48,72 h and 96 h in the group A, group B and group C were significantly increased （P 〈0. 05） , especially, the proliferation ability in the group B was strongest, while the prolifer-ation ability at different points were decreased （P 〈0. 05） . Compared with 24 h,the relative expression levels of A LP ,0C N , and OPN mRNA and the ratios of OPN/RANKL at 48 ,72 h and 96 h in the group A, group B, group C, group D and control group were increased, and showed tendency to increase with time, while the relative expression levels of RANKL mRNA were decreased （ P 〈 0. 05 ） , and showed tendency to decrease with time. Compared among group A, group B, group C,the relative expression levels of ALP, OCN and OPN mRNA and ratios of OPN/RANKL at 48,72 h and 96h were： group B 〉 group C 〉 group A 〉 group D, while the relative expression levels of RANKL mRNA ： group D 〉 group A 〉 group C 〉 group B. Conclusion Leptin could promote proliferation and differentiation of osteoblast-like cells after directional differentiation of BMSCs in postmenopausal osteoporosis patients. This process might be achieved by adjusting the OPN/RANKL pathway,with time-dependent properties, and the optimum concentration.
作者
赵鹏
李树锋
ZHAO Peng LI Shufeng(The Central Hospital of Jiaozuo Coal Group , Jiaozuo, Henan 454000 , China Qianfoshan Hospital of Shandong Province, Jinan,Shandong 250014 , China)
出处
《安徽医药》
CAS
2017年第6期1049-1054,共6页
Anhui Medical and Pharmaceutical Journal
基金
山东省自然科学基金面上项目(ZR2014HM049)
关键词
骨质疏松
绝经后
瘦素
骨髓间质干细胞
机制
Osteoporosis
Postmenopausal
Leptin
Mesenchymal stem cells
Mechanism
