关节软骨细胞线粒体功能障碍与软骨退变的关系

摘要/Abstract

摘要： 目的：探求骨关节炎患者软骨细胞中线粒体功能状况及线粒体功能与软骨退变的关系。方法：收集2012年4月至10月在北京大学第一医院因骨关节炎（osteoarthritis，OA）行膝关节置换术患者的关节软骨10例。根据Outerbridge分级对所取软骨组织进行透射电子显微镜观察。获取正常及OA软骨细胞，用透射电子显微镜观察2组软骨细胞线粒体形态。定量检测正常及OA软骨细胞线粒体呼吸链复合体酶1,2,2+3,4及ATP合酶活力，用JC-1法检测2组软骨细胞线粒体膜电位。获取10例正常软骨细胞并将其分为正常对照组和鱼藤酮处理组，比较2组细胞线粒体超微结构、线粒体膜电位、细胞凋亡及Ⅱ型胶原含量。结果：软骨组织块及软骨细胞电子显微镜均发现病变的软骨细胞线粒体发生肿胀，外膜破裂，嵴破坏、消失。OA软骨细胞线粒体膜电位下降，红绿荧光比为1.50，较正常细胞（2.58）低。OA软骨细胞呼吸链复合酶1,2,2+3,4及ATPase活力都较正常组低，但差异无统计学意义（P=0.109,0.197,0.098,0.169,0.145）。鱼藤酮处理的细胞线粒体出现类似于OA软骨细胞的形态变化。JC-1法示正常软骨细胞红绿荧光比为2.58；鱼藤酮组细胞为1.78。细胞凋亡检测示正常软骨细胞凋亡率为4.38%，鱼藤酮组细胞为7.53%。正常软骨细胞Ⅱ型胶原含量为(72.9±24.3) μg/L，鱼藤酮组为(44.63±7.11) μg/L，较正常组低（P=0.044）。结论： OA软骨细胞中存在线粒体功能障碍，破坏软骨细胞线粒体功能会导致软骨细胞凋亡率增加，分泌Ⅱ型胶原的能力下降，从而促进软骨退变。

关键词: 骨关节炎, 软骨细胞, 线粒体, 鱼藤酮

Abstract: Objective：To evaluate the effect of chondrocyte mitochondrial dysfunction on the development of cartilage degeneration. Methods: In the study, 10 cartilage samples of the knee joint were collected during total knee arthroplasty surgery because of OA from April to October of 2012 in Peking University First Hospital. All the tissues were taken from transmission electron microscope (TEM) observation grouped by Outerbridge classification. Then, TEM observation, quantitative detection of mitochondrial respiratory chain enzyme complex 1,2,2+3,4 and ATPase activity, detection of the mitochondrial membrane potential by JC-1 method were taken with cultured normal and OA chondrocytes. Healthy chondrocytes from 10 normal cartilage samples were divided into 2 groups: the normal control group and rotenone group. The ultrastuctrure alterations of mitochondria, mitochondrial membrane potential, apoptosis rate and collagen Ⅱ content were compared. Results: With the aggravation of cartilage degeneration, mitochondria swelling, outer membrane rupture, cristae destruction and disappearance were observed in both the tissue and cell TEM examinations. JC-1 staining showed a decreased membrane potential in OA chondrocytes which had a lower red/green fluorescence ratio of 1.50 than that of the normal chondrocytes of 2.58. mitochondrial respiratory chain (MRC) enzyme complex 1,2,2+3,4 and ATPase activity of the OA chondrocytes also represented a decreased tendency compared with the normal chondrocytes although the difference was not significant (P=0.109,0.197,0.098,0.169,0.145). The mitochondria in the Ro group cells showed OA-like changes morphologically by TEM detection. JC-1 staining showed a decreased mitochondrial membrane potential in the Ro group chondrocytes which had a lower red/green fluorescence ratio of 1.78 than that of the normal ones of 2.58. Apoptosis examination represented a higher apoptosis rate of 7.53% in the Ro group chondrocytes than that of the normal ones of 4.38%. Collagen Ⅱ content of the chondrocytes in the Ro group was (44.63±7.11) μg/L , significantly lower than (72.88±24.3) μg/L in the control group (P=0.044). Conclusion: Mitochondrial function is impaired in OA chondrocyte. Mitochondrial function destruction results in an increased chondrocyte apoptosis rate and a decreased collagen Ⅱ secretion.

Key words: Osteoarthritis, Chondrocytes, Mitochondria, Rotenone

刘恒, 曹永平, 崔云鹏, 杨昕, 孟志超, 王瑞. 关节软骨细胞线粒体功能障碍与软骨退变的关系[J]. 北京大学学报（医学版）, 2014, 46(5): 760-765.

LIU Heng, CAO Yong-Ping, CUI Yun-Peng, YANG Xin, MENG Zhi-Chao, WANG Rui. Effect of mitochondrial dysfunction of chondrocyte on cartilage degeneration[J]. Journal of Peking University(Health Sciences), 2014, 46(5): 760-765.

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