Huang Zengfa, Cheng Cheng, Cao Beibei, Wang Jing, Wei Hui, Liu Xianzhe, Han Yu, Yang Shuhua, Wang Xiang
Department of Radiology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Cell Physiol Biochem. 2018;47(2):694-706. doi: 10.1159/000490023. Epub 2018 May 22.
BACKGROUND/AIMS: Glucocorticoid (GC)-related osteonecrosis of the femoral head (ONFH) is a common complication following administration of steroids to treat many diseases. Our previous study demonstrated that icariin (ICA) might have a beneficial effect on the bone marrow mesenchymal stem cells (BMSCs) of patients with steroid-associated osteonecrosis. In this study, we investigated the underlying mechanisms of ICA associated with the potential enhancement of osteogenesis and anti-adipogenesis in GC-related ONFH.
In vitro cell proliferation was evaluated by CCK-8 assay. Alizarin red S and alkaline phosphatase (ALP) activity were used to measure osteogenic differentiation, while adipogenic differentiation was revealed by oil red O staining and TG content assay. The expression level of osteogenesis-associated genes and PPARγ was evaluated by RT-qPCR, western blotting and immunofluorescence. A total of 30 female SD rats were randomly separated into three groups: a control group, a methylprednisolone (MPS) group and a MPS + ICA group. Serum ALP and TG (triglyceride), micro-CT scanning, histological and immunohistochemical analyses were performed in the animal model.
In the in vitro study, ICA promoted proliferation, improved osteogenic differentiation and suppressed adipogenic differentiation of BMSCs treated with MPS. The group treated with MPS and 10-6 M ICA expressed higher levels of Runx2, ALP, bone morphogenetic protein (BMP) 2, and OC and lower expression of PPARγ than the MPS group. In the in vivo study, ICA prevented bone loss in a rat model of GC-related ONFH as shown by micro-CT scanning, histological and immunohistochemical analyses.
ICA is an effective compound for promoting bone repair and preventing or delaying the progression of GC-associated ONFH in rats. This effect can be explained by its ability to improve the balance between adipogenesis and osteogenesis, indicating that ICA is an effective candidate for management of GC-associated ONFH.
背景/目的：糖皮质激素（GC）相关的股骨头坏死（ONFH）是使用类固醇治疗多种疾病后常见的并发症。我们之前的研究表明，淫羊藿苷（ICA）可能对类固醇相关性骨坏死患者的骨髓间充质干细胞（BMSC）具有有益作用。在本研究中，我们探讨了ICA与GC相关ONFH中潜在的成骨增强和抗脂肪生成相关的潜在机制。
通过CCK-8法评估体外细胞增殖。茜素红S和碱性磷酸酶（ALP）活性用于测量成骨分化，而油红O染色和甘油三酯（TG）含量测定用于揭示脂肪生成分化。通过RT-qPCR、蛋白质免疫印迹和免疫荧光评估成骨相关基因和PPARγ的表达水平。将30只雌性SD大鼠随机分为三组：对照组、甲基泼尼松龙（MPS）组和MPS + ICA组。在动物模型中进行血清ALP和TG（甘油三酯）、显微CT扫描、组织学和免疫组织化学分析。
在体外研究中，ICA促进了用MPS处理的BMSC的增殖，改善了成骨分化并抑制了脂肪生成分化。与MPS组相比，用MPS和10-6 M ICA处理的组Runx2、ALP、骨形态发生蛋白（BMP）2和OC的表达水平更高，而PPARγ的表达更低。在体内研究中，显微CT扫描、组织学和免疫组织化学分析表明，ICA可预防GC相关ONFH大鼠模型中的骨质流失。
ICA是促进大鼠骨修复、预防或延缓GC相关ONFH进展的有效化合物。这种作用可以通过其改善脂肪生成和成骨之间平衡的能力来解释，表明ICA是治疗GC相关ONFH的有效候选药物。
