Phenotype changes of subchondral plate osteoblasts based on a rat model of ovariectomy-induced osteoarthritis
Phenotype changes of subchondral plate osteoblasts based on a rat model of ovariectomy-induced osteoarthritis
复制标题
基于卵巢切除诱发骨关节炎大鼠模型的软骨下板成骨细胞表型变化
DOI：
10.21037/atm.2020.03.93
复制
发表时间：
2020-04-01
影响因子：
--
通讯作者：
Leng, Huijie
中科院分区：
文献类型：
--
作者：
Jiang, Ai;Gao, Shan;Leng, Huijie
骨关节炎（OA）是绝经后妇女的常见病。与其他类型的OA相比，去卵巢（OVX）大鼠的软骨下骨可能在软骨退变中起更重要的作用。OVX大鼠软骨下成骨细胞如何变化仍不清楚。了解OVX软骨下骨成骨细胞的变化可能有助于阐明OVX-OA的发病机制。方法雌性SD大鼠随机分为假手术组（n=20）和去卵巢组（n=20）。每1 ~ 2周检测血清碱性磷酸酶（ALP）和I型胶原C端肽（CTX-I）水平。所有大鼠在术后第9周处死。测定大鼠体重和子宫湿重。采用显微计算机断层扫描（micro-CT）分析膝关节显微结构，甲苯胺蓝染色评价软骨侵蚀。采用kit-8细胞计数法检测Sham组和OVX组软骨下成骨细胞的增殖能力，逆转录聚合酶链反应（RT-PCR）检测成骨基因的表达，碱性磷酸酶（ALP）染色和茜素红染色检测成骨细胞的分化和矿化能力。结果卵巢切除后第2周血清ALP和CTX-I水平明显升高。术后第9周，去卵巢大鼠体重明显增加，子宫重量明显下降。与Sham大鼠相比，OVX大鼠表现出显著的软骨下骨改变和软骨侵蚀。成骨分化早期标志物（ALP、I型胶原、Runx 2）mRNA水平在OVX大鼠中增强，但晚期标志物（骨钙素）无显著差异。去卵巢大鼠成骨细胞ALP活性升高，矿化能力下降。结论去卵巢大鼠软骨下成骨细胞的增殖、分化和矿化能力与正常对照组不同。
Background Osteoarthritis (OA) is prevalent in postmenopausal women. Subchondral bone in ovariectomized (OVX) rats might play a more important role in cartilage degeneration compared with other types of OA. How subchondral osteoblast changes in OVX rats is still unclear. Understanding of osteoblast changes obtained from OVX subchondral bone might be helpful to clarify pathogenesis of OVX-OA. Methods Female Sprague-Dawley rats were randomly divided into two groups: Sham (n=20) and OVX (n=20). Serum levels of Alkaline phosphatase (ALP) and C-telopeptide of type I collagen (CTX-I) were measured every one or two weeks. All rats were executed at week 9 post surgery. The weight of rats and the wet weight of uterus were assessed. Micro-computed Tomography (micro-CT) was used to analyze the knee microstructure, and toluidine blue staining was employed to evaluate cartilage erosion. Subchondral osteoblast proliferation ability by cell counting kit-8 assay, osteogenic genes expressions by reverse transcription polymerase chain reaction (RT-PCR), differentiation and mineralization ability by ALP staining and alizarin red staining were evaluated and compared between Sham and OVX. Results Ovariectomy induced significant increases of serum ALP and CTX-I as early as at week 2. At week 9 after surgery, the body weight of OVX rats was significantly increased, and uterus weight of OVX rats was remarkably decreased. OVX rats demonstrated significant subchondral bone change and cartilage erosion compared with Sham rats. mRNA levels of early markers of osteogenic differentiation (ALP, type I collagen, Runx2) were enhanced in OVX rats, but the late marker (osteocalcin) was not significantly different. ALP activity of osteoblasts increased, but the mineralization capacity decreased in OVX rats. Conclusions Subchondral osteoblasts in OVX rats exhibited different proliferation, differentiation and mineralization abilities from normal counterparts.