AGEs诱导软骨细胞凋亡和影响细胞外基质变化的研究

Advanced Glycation End Products induced apoptosis and expression of extracellular matrix in mouse chondrocytes

目的 :明确晚期糖基化终末产物(advanced glycation end products,AGEs)诱导小鼠关节软骨细胞凋亡情况和影响细胞外基质变化。方法:体外分离培养小鼠软骨关节细胞,给予不同浓度AGEs共培养后,分别于24 h和48 h时间点观察细胞凋亡情况;通过Western blot方法检测Ⅰ型胶原、MMP-3、MMP-9以及p53蛋白的表达情况。结果:不同浓度AGEs处理软骨细胞24 h后,与对照组比较,细胞凋亡率出现升高(P<0.05,n=3);与BSA组比较,200 mg/L浓度的AGEs组细胞凋亡率明显升高(P<0.05,n=3),并且随着AGEs浓度和培养时间增加,软骨细胞的凋亡率也增加。200 mg/L的浓度AGEs处理软骨细胞48 h后,Ⅰ型胶原蛋白的表达显著下降(P<0.05,n=3);另外,50、100、200 mg/L的浓度AGEs处理软骨细胞48 h后,p53、MMP-3、MMP-9的表达显著高于对照组和BSA组(P<0.05,n=3)。结论 :AGEs能诱导小鼠软骨细胞凋亡,并刺激软骨细胞p53、MMP-3、MMP-9表达增多,降低Ⅰ型胶原蛋白表达。

Objective：To investigate the effects of advanced glycation end products（AGEs） on apoptosis and approach the mechanism. Methods：The primary cultured mouse chondrocytes were incubated with different concentration of AGEs for 24 h or 48 h. Apoptosis of chondrocytes was detected by FCW ;expression of Col Ⅰ , MMP-3, MMP-9 and p53 was measured by Western blot. Results：After the chondrocytes were treatment with AGEs （50,100,200 mg/L） for 24 h,the apoptosis was increased compared with the control groups;the maximum stimulation of chondrocytes was 200 mg/L AGEs. After the chondrocytes were treatment with AGEs （50,100,200 mg/L） for 48 h, expression of Col was significamtly decreased （P〈0.05,n=3） while the levels of MMP-3, MMP-9 and p53 were increased,compared to control groups and BSA group. Conclusion：AGEs can increase the apoptosis of mouse chondrocytes in a dose-dependent manner, thus damage in chondrocytes via decrease of the expression of extracellular matrix.