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| 胶原蛋白酶诱发兔膝关节骨性关节炎软骨组织中HIF2α表达上调 |
| Up-regulation of HIF2α in collagenase-induced knee osteoarthritis in rabbits |
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| DOI：10.3969/j.issn.1006.7108.2015.11.005 |
| 中文关键词: 骨性关节炎 膝关节 软骨 HIF2α |
| 英文关键词:Osteoarthritis Knee joint Cartilage HIF2α |
| 基金项目:国家自然科学基金（81330043);国家自然科学基金（81472139);北京市优秀人才青年骨干个人项目（2014000021469G221） |
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| 中文摘要: |
| 目的 研究低氧诱导因子HIF2α在胶原蛋白酶诱发兔膝关节骨性关节炎（Osteoarthritis, OA )模型中的表达情况，为下一步以HIF2α为靶点进行OA靶向治疗研究提供实验基础。方法 取健康16周龄雄性日本大耳白兔24只，随机分为磷酸盐缓冲液（PBS)对照组和胶原蛋白酶诱发OA ( Collagenase-induced OA, CIOA)组。利用右侧膝关节腔内注射II型胶原蛋白酶 (剂量为0. 5 mg)方法建立膝关节OA模型。给药后24周处死动物,采用大体形态观察与HE、甲苯胺蓝染色观察关节软骨退变程度，应用微型CT( Micro-CT)观察软骨下骨改变情况，采用免疫组织化学（immunohistochemistry ,IHC)、蛋白质免疫印迹 (Western Wotting, WB)和实时荧光定量PCR( quantitative real time PCR, qRT-PCR)检测软骨中 HIF2α 表达情况。结果 大体形态观察结果显示，CIOA组关节面粗糙、糜烂，与溃疡形成,关节囊增厚，滑膜增生。组织病理学检测结果显示，CIOA组关节软骨缺损，软骨细胞数量减少，排序紊乱，有细胞簇聚现象。Micro-CT结果显示，CIOA组软骨下骨可见大量骨赘形成,关节间隙狭窄。IHC结果显示，CIOA组关节软骨中HIF2α蛋白阳性细胞数增加；WB结果显示，CIOA组关节软骨组织中HIF2α蛋白表达显著增加；而qRT-PCR结果显示，HIF2α mRNA表达显著降低。结论 在成功建立膝关节OA模型基础上，发现HIF2α在胶原蛋白酶诱发兔膝关节OA软骨中表达上调，其表达调控可能在转录后水平进行。我们的实验结果提示HIF2α特异性抑制剂可能被用于缓解OA发生。 |
| 英文摘要: |
| Objective To study the expression of hypoxia-inducible factor 2α (HIF2α) in a rabbit collagenase-induced knee osteoarthritis (OA) model. The study may provide experimental evidence that HIF2α serves as a therapeutic target for the treatment of OA of the knee in the future. Methods A total of 24 16-week healthy male Japanese white rabbits with long ears were randomly divided into collagenase-induced OA (CIOA) group and phosphate buffered saline (PBS) control group. Rabbits received intra-articular injection of 0.5 mg type II collagenase to establish OA. After 24 weeks of injection, the rabbits were sacrificed. The severity of the articular cartilage degeneration was observed using HE and toluidine blue staining. The subchondral bone was observed using micro-CT. The expression of HIF2α in articular cartilage was detected using immunohistochemistry (IHC), Western blotting (WB)，and quantitative real time PCR (qRT-PCR). Results Macroscopic results showed a spectrum of lesions in the knee joints in CIOA group，including superficial and deep clefts, erosion, and ulceration of articular cartilage, thickening of the joint capsule, and synovial hyperplasia, Histopathological examination confirmed the loss of articular cartilage of CIOA group, showing a decrease in the number and organization of chondrocytes, and the formation of chondrocyte clusters. Micro-CT demonstrated osteophyte formation and joint space narrowing in the collagenase-treated knee joints. The IHC results showed an increase in the number of HIF2α-positive chondrocytes. WB results showed an increase of HIF2α protein in the articular cartilage of collagenase-treated knee joints. The mRNA level of HIF2α decreased as detected by qRT-PCR. Conclusion HIF2α is up-regulated in a rabbit model of collagenase-induced knee OA, and its expression is regulated at the post-transcriptional level. Our results suggest that HIF2α inhibitors should be applied to treat knee OA. |
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