Role of mesenchymal stem cells on differentiation in steroid-induced avascular necrosis of the femoral head
Role of mesenchymal stem cells on differentiation in steroid-induced avascular necrosis of the femoral head
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DOI：
10.3892/etm.2016.3991
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发表时间：
2017-02-01
影响因子：
2.7
通讯作者：
Guo, Li
中科院分区：
文献类型：
--
作者：
Wang, Tiansheng;Teng, Shoufa;Guo, Li
已知类固醇抑制间充质干细胞（MSC）的成骨分化和减少骨形成，同时伴随诱导类固醇诱导的股骨头缺血性坏死（SANFH）。本研究的目的是评估骨髓间充质干细胞在SANFH分化中的作用，并在兔模型中研究SANFH的病理生物学机制。对照组、创伤诱导ANFH（TANFH）组和SANFH组的MSC在含10%胎牛血清的低糖完全Dulbecco改良Eagle培养基中孵育。用苏丹III染色MSC中的脂肪细胞的数量，并使用光学显微镜计数。分别采用定量聚合酶链反应和蛋白质印迹分析法测定脂肪特异性422（AP 2）、过氧化物酶体增殖物激活受体-γ（PPAR γ）、RUNX 2、I型胶原（Col I）和miR-103在MSC中的mRNA和蛋白质表达水平。采用放射免疫分析法和试剂盒检测骨髓间充质干细胞骨钙素（OC）、碱性磷酸酶（ALP）和甘油三酯（TG）的活性。在SANFH组的MSC中，AP 2和PPAR γ的mRNA和蛋白表达水平增加，而RUNX 2和Col I的mRNA和蛋白表达水平降低。SANFH组MSCs中OC和ALP活性降低，TG活性升高。此外，SANFH组MSCs中miR-103的表达升高。常规培养3周后，SANFH组MSC群中脂肪细胞的数量增加。因此，本研究结果提示，SANFH中MSCs的成骨分化减弱，脂肪分化增强，这为SANFH相关的病理变化提供了新的解释。
Steroids are known to inhibit osteogenic differentiation and decrease bone formation in mesenchymal stem cells (MSCs), while concomitantly inducing steroid-induced avascular necrosis of the femoral head (SANFH). The aim of the present study was to evaluate the function of MSCs on differentiation in SANFH and investigate the pathobiological mechanisms underlying SANFH in a rabbit model. MSCs in the control, trauma-induced ANFH (TANFH) and SANFH groups were incubated with low-glucose complete Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. A number of adipocytes in the MSCs were stained with Sudan III and counted using a light microscope. The mRNA and protein expression levels of the adipose-specific 422 (AP2), peroxisome proliferator-activated receptor-gamma (PPAR gamma), RUNX2, collagen type I (Col I) and miR-103 in the MSCs were determined using quantitative polymerase chain reaction and western blot analysis, respectively. In addition, the activities of osteocalcin (OC), alkaline phosphatase (ALP) and triglyceride (TG) in MSCs were analyzed using radioimmunoassay and determination kits. In the MSCs of the SANFH group, the mRNA and protein expression levels of AP2 and PPAR gamma were increased, while those of RUNX2 and Col I were reduced. Furthermore, the levels of OC and ALP activity in the MSCs of the SANFH group were decreased, and the activity of TG in the MSCs of the SANFH group was increased. In addition, the expression of miR-103 in the MSCs of the SANFH group was elevated. Following routine culture of the MSCs for 3 weeks, the number of adipocytes among the MSC population of the SANFH group was increased. Therefore, the results of the present study suggest that the osteogenic differentiation of MSCs in the SANFH was mitigated, while fat differentiation was promoted, which provides a novel explanation for the pathological changes associated with SANFH.