内质网应激对膝骨关节炎大鼠关节软骨细胞的影响

发布时间：2018-12-05 19:03

【摘要】：背景:研究认为,内质网应激与糖尿病、扩张性心肌病、神经退行性疾病等许多慢性疾病的发生相关,而它与骨关节炎的发生也密切相关。目的:分析内质网应激对大鼠关节软骨细胞生存状态的影响,以及内质网应激在体内对大鼠骨关节炎发生发展的影响。方法:在体外分离培养大鼠关节软骨细胞,应用10 mg/L衣霉素建立内质网应激模型。Western Blot检测内质网应激相关蛋白葡萄糖调节蛋白78和C/EBP同源蛋白的表达,并分别应用CCK-8和Annexin V-FITC(流式法)检测软骨细胞增殖活性及细胞凋亡情况。体内实验选取SD大鼠15只,行前交叉韧带切断、内侧半月板切除术制作膝关节骨关节炎模型,于关节腔分别注射内质网应激激动剂衣霉素、抑制剂牛磺熊去氧胆酸和空白对照PBS,4周后应用苏木精-伊红染色评估各组关节退变情况。结果与结论:(1)应用衣霉素刺激大鼠关节软骨细胞后,葡萄糖调节蛋白78和C/EBP同源蛋白的表达均显著升高;同时,软骨细胞的细胞增殖活性在衣霉素刺激后逐渐下降,而细胞凋亡率则明显升高;(2)在大鼠骨关节炎模型中,大体标本及苏木精-伊红染色结果均显示,关节腔注射衣霉素组骨关节炎进展加速,而关节腔注射牛磺熊去氧胆酸组骨关节炎进展减慢;(3)结论:内质网应激的过度激活导致体外培养的大鼠关节软骨细胞增殖活性降低及细胞凋亡增加,这可能是导致骨关节炎发生发展的重要机制之一;应用内质网应激抑制剂牛磺熊去氧胆酸可以有效地减少内质网应激导致的骨关节炎的进展。
[Abstract]:Background: endoplasmic reticulum stress is associated with many chronic diseases such as diabetes mellitus dilated cardiomyopathy neurodegenerative diseases and osteoarthritis. Aim: to analyze the effect of endoplasmic reticulum stress on the survival state of rat articular chondrocytes and the effect of endoplasmic reticulum stress on the occurrence and development of osteoarthritis in vivo. Methods: rat articular chondrocytes were isolated and cultured in vitro. Endoplasmic reticulum stress (ER) stress model was established by 10 mg/L. Western Blot to detect the expression of endoplasmic reticulum stress-related protein glucose-regulated protein 78 and C/EBP homologous protein. The proliferative activity and apoptosis of chondrocytes were detected by CCK-8 and Annexin V-FITC. In vivo, 15 SD rats were treated with anterior cruciate ligament transection and medial meniscectomy to make the osteoarthritis model of knee joint, and the endoplasmic reticulum stress agonist, chlamycin, was injected into the articular cavity. The deoxycholic acid (Taurodeoxycholic acid) inhibitor and blank control group (PBS,4) were used to evaluate joint degeneration by hematoxylin-eosin staining. Results and conclusion: (1) the expression of glucose-regulated protein 78 and C/EBP homologous protein increased significantly in articular chondrocytes of rats stimulated by itamycin. At the same time, the proliferation activity of chondrocytes decreased gradually after stimulation of itamine, while the apoptosis rate increased significantly. (2) in the rat model of osteoarthritis, gross specimens and hematoxylin-eosin staining showed that the progress of osteoarthritis was accelerated in the group of intraarticular injection of itamycin, but the progression of osteoarthritis was slowed down in the group of intraarticular injection of ursodeoxycholic acid. (3) conclusion: the excessive activation of endoplasmic reticulum stress leads to the decrease of proliferation activity and the increase of apoptosis in cultured rat articular chondrocytes, which may be one of the important mechanisms leading to the occurrence and development of osteoarthritis. Endoplasmic reticulum stress inhibitor taurodeoxycholic acid can effectively reduce the progress of osteoarthritis induced by endoplasmic reticulum stress.
【作者单位】：北京大学第一医院;
【基金】：北京市自然科学基金青年基金项目(7144252)~~
【分类号】：R684.3

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