摘要
目的研究免疫抑制剂FKS06在体外对大鼠骨髓来源的间充质干细胞(MSCs)成骨诱导能力以及FK506成骨诱导的量效关系。方法MSCs在不同条件下培养16d：(1)对照组(C组)- (基础培养液)：含L-抗坏血酸-2-磷酸(AsAp)和β-甘油磷酸(β-GP)的α-MEM培养液；(2)基础培养液加入地塞米松(D组)；(3)基础培养液加入FK506(F组)；(4)基础培养液加入地塞米松和FKS06(FD组)。通过测定干细胞形态、细胞增殖、碱性磷酸酶(ALP)活性、钙化结节染色和骨钙素基因表达等,评价FK506对MSCs的成骨诱导能力。结果FK506能促进大鼠MSCs增殖、成骨细胞分化,明显诱导钙化结节形成,显著提高ALP活性和骨钙素mRNA的表达。FK506与地塞米松具有协同成骨诱导作用,当α-MEM中含0．25mmol/L AsAp、10mmol/Lβ-甘油磷酸、10nmol/L地塞米松和50nmol/L FK506时可获得最佳成骨效应。结论FK506具有强大成骨诱导能力,可以用作骨诱导因子。
Objective To investigate the effect of FKS06 on the in vitro osteogenic potential of rat lone marrow-derived mesenchymal stem cells （MSCs） and the dose-response effect of FKS06 （5 -5000 nmol/ L） on the osteogenic potential of MSCs in vitro. Methods MSCs derived from primary culture were subcultured for 16 days under four conditions： （ 1 ） α-MEM containing L - ascorbieacid-2-phosphate （AsAP） and β-glycerophosphate （β-GP） （basic cuhure medium） as a control; （2） AsAP and β-GP （basic culture medi- um） plus dexamethasone ; （3） AsAP and β-GP （basic culture medium ） plus FK506, （4） AsAP and β-GP （basic culture medium ） plus FK506 and dexamethasone. Osteogenic potential was determined by testing osteblastic morphology, cell proliferation, alkaline phosphatase （APase） activity, lone nodule formation and the expression of osteocalcin mRNA. Results FK506 promoted the proliferation of MSCs, induced mineralizing bone-like nodule formation, and increaased APase activity and the expression of osteocalcin mRNA. The data also showed that the efficacy of FK506 was greater when used in combination with dexamethasone. Optimal osteogenesis was achieved with α-MEM containing 0. 25mmol/L AsAP, 10mmol/L β-GP, 10nmol/L dexamethasone and 50nmol/L FK506. Conclusion FK506 has powerful osteogenic ablility. It can be considered as an osteogenic agent to repair bene defects.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2006年第10期775-778,共4页
Chinese Journal of Trauma
基金
国家自然科学基金(30271262)
上海市卫生局科技发展基金(054076)
