甲泼尼龙对鸡关节软骨细胞自噬的影响

Effects of methylprednisolone on chicken chondrocyte autophagy

为探究甲泼尼龙(MP)对关节软骨细胞自噬的影响,机械分离肉鸡膝关节软骨组织,剪碎后,0.25%胰蛋白酶消化30 min,在0.1%的透明质酸酶和0.2%的Ⅱ型胶原酶中消化12 h,于含10%胎牛血清的培养液中进行细胞培养。采用甲苯胺蓝、阿利新蓝染色法和PCR扩增鉴定关节软骨细胞,采用MP和三甲基腺嘌呤(3-MA)处理鸡关节软骨细胞。结果显示,MP促进软骨细胞自噬相关基因的显著表达,并降低细胞活性,3-MA对MP诱导的关节软骨细胞自噬有保护作用。本研究建立了原代肉鸡关节软骨细胞培养方法,初步揭示了MP及3-MA对软骨细胞自噬的作用机理。

This study aimed to investigate the effects of methylprednisolone(MP)on articular chondrocyte autophagy.Articular cartilage tissue from broilers’knee-joint was mechanically isolated,cut into pieces,digested with 0.25%trypsin for 30 min,and then 0.1%hyaluronidase and 0.2%collagenase typeⅡfor 12 h.The culture medium supplemented with 10%fatal bovine serun(FBS)was used for the cell culture.The articular chondrocytes were identified via toluidine blue,alcian blue staining,and PCR amplification respectively.Articular chondrocytes were treated with MP and 3-methyladenine(3-MA).The results showed that MP significantly promoted the expressions of autophagy-related genes in chondrocytes and decreased the cell viability,furthermore,3-MA had a protective effect on MP-induced autophagy in articular chondrocytes.This study established a method of primary chicken articular chondrocyte culture,and preliminarily revealed the mechanism of MP and 3-MA on chondrocyte autophagy.