目的:探讨自噬在骨关节炎(OA)软骨细胞中的表达及意义。方法收集临床 OA 及正常骨关节软骨标本各6例,分为 OA 组和对照组。免疫组化法检测自噬相关蛋白微管相关蛋白轻链3(LC3)和 Beclin-1在软骨组织中表达情况。分离关节软骨细胞并在低氧环境下培养,采用瞬时转染绿色荧光蛋白 LC3(GFP-LC3)在激光共聚焦显微镜下观察各组软骨细胞自噬变化,并用 Western blot 法检测软骨细胞中 LC3蛋白表达情况。结果 OA 组软骨组织中 LC3蛋白和Beclin-1蛋白的表达显著高于对照组(P <0.05);在 OA 组发现大量自噬颗粒形成,OA 组 GFP-LC3细胞阳性率显著高于对照组;OA 组软骨细胞中 LC3蛋白从 LC3-Ⅰ向 LC3-Ⅱ转换表达显著高于对照组。结论自噬在 OA 软骨细胞退变的过程中起着重要作用,为 OA 发病机制的研究提供新的方向。
Objective To explore the autophagy expression and examine its significance in chondrocytes from a de-generate of osteoarthritis (OA). Methods Cartilage samples were obtained from 12 hospitalized patients in our hospital. They were divided into the OA group (n = 6) and the control group (n = 6). Expressions of light chain3 (LC3) and Beclin-1 protein which were main markers of autophagy were analyzed in cartilage tissues by using im-munohistochemistry (IHC). Cartilage chondrocytes were isolated and cultured in a hypoxia environment in vivo. The chondrocytes which were transfected with green fluorescent protein-light chain3 (GFP-LC3) plasmid were ex-amined under a laser scanning confocal microscopy. Western blot was used to observe the LC3 protein expression in chondrocytes. Results In cartilage tissues LC3 and Beclin-1 protein expressions were markedly increased in OA chondrocytes. A large number of autophagy particle formations were found in the OA group, the percentage of chon-drocytes with GFP-LC3 puncta structures was higher in the OA group than the control group. The results revealed a significant increase in the expression of LC3-II protein in the OA group. Conclusion Autophagy plays an impor-tant pathogenic role in the process of OA chondrocytes degeneration. It offers a new research direction for the patho-genesis of OA.