目的 探讨高糖对大鼠心肌成纤维细胞Ⅰ、Ⅲ型胶原表达的影响.方法 采用胰酶消化法和差速贴壁分离法获取心肌成纤维细胞,采用正常糖(5.5 mmol/L,对照组)和高糖(25 mmol/L,高糖组)干预.用实时荧光定量PCR(Real-time PCR)方法检测Ⅰ、Ⅲ型前胶原mRNA表达,酶联免疫吸附测定法(ELISA)观察Ⅰ、Ⅲ型胶原蛋白水平的变化.结果 与对照组比较,高糖刺激12 h可使心肌成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达增加,Ⅰ、Ⅲ前胶原mRNA表达在12 h升高,在24 h达高峰,Ⅰ型前胶原mRNA升高幅度高于Ⅲ型前胶原mRNA;高糖刺激48 h可使Ⅰ、Ⅲ型胶原蛋白表达增加,Ⅰ型胶原蛋白升高幅度高于Ⅲ型胶原.结论 高糖可促进大鼠心肌成纤维细胞合成Ⅰ、Ⅲ型胶原.%Objective To investigate the effect of high glucose on the expression of type Ⅰ and Ⅲ collagen in rat cardiac fibroblasts .Methods Rat cardiac fibroblasts were obtained by pancreatic enzyme digestion method and the differential sidewall separation method .Normal glucose (5 .5 mmol/L, control group) and high glucose(25 mmol/L, high glucose group) were adopted for conducting intervention .The mRNA expressions of type Ⅰ and Ⅲ procollagen were detected by real-time PCR and the changes of type Ⅰ and Ⅲ collagen levels were detected by ELISA .Results Compared with the control group ,the high glucose 12 h stimulation could increase the mRNA expression of Ⅰ and Ⅲ procollagen in rat cardiac fibroblasts .The mRNA expression of collagen was increased at 12 h and reached the peak at 24 h .The increasing amplitude of type Ⅰ collagen mRNA was higher than that of type Ⅲ collagen .The high glucose 48 h stimulation could increase the expression of type Ⅰ and Ⅲ collagen .The increasing amplitude of type Ⅰ collagen was higher than that of collagen type Ⅲ.Conclusion High glucose could promote the synthesis of type Ⅰ and Ⅲ collagen in rat cardiac fibroblasts .
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