来源：风湿病与关节炎,2021,10（4）:1-4,31.

自噬在骨关节炎软骨细胞退变中表达的研究（基础研究）

王　浩，陈群超，殷丽丽

　　【摘　要】目的：探究不同退变阶段骨关节炎自噬因子ULK1、Beclin1和LC3B在软骨细胞中的表达。方法：根据Kellgren-Lawrence影像学诊断标准，收集正常软骨样品，中期、晚期骨关节炎软骨样品；体外分离、培养软骨细胞，甲苯胺蓝和Ⅱ型胶原免疫组化染色用于鉴定软骨细胞，运用荧光定量PCR及WB技术分别从mRNA及蛋白水平检测ULK1、Beclin1和LC3B的表达。结果：甲苯胺蓝染色显示，软骨细胞核为深蓝色，胞质中有大量异色蓝紫色颗粒。胶原蛋白的免疫荧光染色显示，细胞外基质为棕色，细胞核染为黄褐色。荧光定量PCR及WB结果显示，正常软骨细胞中ULK1、Beclin1、LC3B mRNA及蛋白水平稳定表达，随着骨关节炎程度加重，其表达量逐渐减少，细胞中表达量最低的是晚期骨关节炎组，3组间比较，差异有统计学意义（P < 0.05）。结论：随着骨关节炎的加重，软骨细胞自噬相关因子的表达降低。自噬是维持正常软骨细胞稳态的保护机制，对调控骨关节炎的发展有重要的作用。

　　【关键词】　骨关节炎；软骨细胞；自噬；退变

Study on the Expression of Autophagy in Osteoarthritis Chondrocyte Degeneration

WANG Hao,CHEN qun-chao,YIN Li-li

　　【ABSTRACT】Objective:To investigate the expressions of autophagy factors ULK1,Beclin1 and LC3B in chondrocytes of osteoarthritis at different stages of degeneration.Methods:According to the Kellgren-Lawrence imaging diagnostic criteria,normal cartilage samples,mid-term and late osteoarthritis cartilage samples were collected.Chondrocytes were isolated and cultured in vitro;toluidine blue and typeⅡ collagen immunohistochemical staining were used to identify chondrocytes;and the expressions of ULK1,Beclin1 and LC3B were detected by fluorescent quantitative PCR and WB Technology respectively from mRNA and protein levels.Results:Toluidine blue staining showed that the nucleus of cartilage was dark blue,and there were a large number of heterochromatic blue purple granules in the cytoplasm.Immunofluorescence staining of collagen showed that the extracellular matrix was brown and the nucleus was yellowish brown.The results of fluorescence quantitative PCR and WB showed that ULK1,Beclin1 and LC3B were stably expressed in normal chondrocytes at mRNA and protein levels.With the aggravation of osteoarthritis,the expression level gradually decreased.The lowest expression level was in the advanced osteoarthritis group,and the difference was statistically significant

（P < 0.05）.Conclusion:With the aggravation of osteoarthritis,the expression of autophagy related factors in chondrocytes decreased.Autophagy is a protective mechanism to maintain normal chondrocyte homeostasis,and plays an important role in regulating the development of osteoarthritis.

　　【Keywords】 osteoarthritis;chondroc-yte;autophagy;degeneration