CAJ | 학술논문

目的：研究ASIC1a(acid-sensing ion channel 1a)对大鼠关节软骨细胞基质代谢及MAPK信号通路表达的影响。方法 SD大鼠关节软骨细胞分离、培养与鉴定,建立软骨细胞ASIC1a 表达沉默模型。将软骨细胞分为正常组( pH 7.4)、pH 6.0酸化组、以及ASIC1a特异性阻滞剂PcTx-1、表达沉默组和非特异性阻滞剂Amiloride处理的酸化组,对二甲基亚甲蓝分光法检测大鼠关节软骨细胞培养上清中GAG的含量,氯胺T法检测 Hyp的含量, ELISA法检测 MMP-2、TIMP-2的含量,Western blot 法检测酸化及阻断 ASIC1a 后ERK1/2、p38 MAPK 磷酸化蛋白的表达。结果激活ASIC1a明显抑制大鼠关节软骨细胞GAG、Hyp和TIMP-2代谢水平( P<0.01),对MMP-2的代谢水平降低抑制作用较弱(P<0.01),且ASIC1a能引起ERK1/2、p38 MAPK磷酸化水平升高,阻断 ASIC1a后,磷酸化水平升高受到明显抑制(P<0.01)。结论 ASIC1a参与了酸诱导的大鼠关节软骨细胞基质代谢的失衡,其机制可能与激活 ERK1/2和 p38 MAPK磷酸化有关。
목적：연구ASIC1a(acid-sensing ion channel 1a)대대서관절연골세포기질대사급MAPK신호통로표체적영향。방법 SD대서관절연골세포분리、배양여감정,건립연골세포ASIC1a 표체침묵모형。장연골세포분위정상조( pH 7.4)、pH 6.0산화조、이급ASIC1a특이성조체제PcTx-1、표체침묵조화비특이성조체제Amiloride처리적산화조,대이갑기아갑람분광법검측대서관절연골세포배양상청중GAG적함량,록알T법검측 Hyp적함량, ELISA법검측 MMP-2、TIMP-2적함량,Western blot 법검측산화급조단 ASIC1a 후ERK1/2、p38 MAPK 린산화단백적표체。결과격활ASIC1a명현억제대서관절연골세포GAG、Hyp화TIMP-2대사수평( P<0.01),대MMP-2적대사수평강저억제작용교약(P<0.01),차ASIC1a능인기ERK1/2、p38 MAPK린산화수평승고,조단 ASIC1a후,린산화수평승고수도명현억제(P<0.01)。결론 ASIC1a삼여료산유도적대서관절연골세포기질대사적실형,기궤제가능여격활 ERK1/2화 p38 MAPK린산화유관。
Aim TostudytheroleofASIC1aonthe matrix turnover and MAPK expression of the rat articu-lar chondrocytes with extracellular acidosis. Methods ArticularchondrocyteswereisolatedfromSprague-Dawley rats, and their phenotype was determined by toluidine blue and immunocytochemical staining. The GAG content of cell culture supernatant was deter-mined by dimethyl-methylene blue spectrophotometric assay, while Hyp content by chloramine T assay. ELISA assay was used to measure MMP-2 , TIMP-2 content. Furthermore, the ERK1/2, p38 MAPK phos-phorylation protein expression levels were tested by Westernblotassay.Results ASIC1acontributedto the effect of GAG, Hyp and TIMP-2 levels reduction induced by extracellular acidification, while the effect of MMP-2 was weaker. Moreover, ASIC1a could in-crease the ERK1/2 , p38 MAPK phosphorylation pro-teinexpressionlevels.Conclusion ASIC1acould regulate rat articular chondrocytes matrix turnover via ERK1/2 and p38 MAPK signaling pathway, and there-by inhibit the rat articular cartilage damage induced by acidosis.