摘要目的 探讨黄芪皂甙Ⅳ(XGA)对大鼠心肌成纤维细胞胶原影响及其量效和时效关系.方法 采用胶原酶(胰蛋白酶消化法分离大鼠心肌成纤维细胞(FBC),建立FBC培养系统.在FBC培养系统内加入不同质量浓度和不同作用时间的XGA,提取RNA后用反转录PCR(RT-PCR)法检测Ⅰ、Ⅲ、Ⅳ型胶原,基质金属蛋白酶(MMP-1、-2、-9)及其抑制剂(TIMP-1、-2)mRNA的表达水平.结果 予不同水平和不同作用时间XGA后, FBC培养系统RT-PCR产物凝胶电泳显示有Ⅰ、Ⅲ、Ⅳ型胶原,MMP-1、-2、-9、TIMP-1和-2 mRNA表达;与予XGA前比较,Ⅰ、Ⅲ、Ⅳ型胶原,TIMP-1、-2 mRNA表达水平有所下降,而MMP-1、-2、-9 mRNA表达水平有所上升,且随着XGA剂量增加或作用时间延长而渐下降或上升.Ⅰ、Ⅲ、Ⅳ型胶原,TIMP-1、-2 mRNA表达水平与XGA的剂量和作用时间呈负相关(r=-0.927～-0.637 P=0～0.024); MMP-1、-2、-9 mRNA表达水平与XGA剂量和作用时间呈正相关(r=0.672～0.962 P=0～0.034).结论 XGA可减少心肌成纤维细胞胶原形成,其机制可能为抑制胶原的合成、增加胶原降解.

abstractsObjective To investigate the dose- and time-effects of astragaloside Ⅳ(XGA) on collagen of myocardial fibroblasts in rats.Methods The myocardial fibroblasts of rats were separated by collagenase and trypsinase digestive method,and the cell culture system was established. After XGA in different concentrations and at different time points was administered in fibroblast culture systems,the mRNA expression levels of collagen,matrix metalloproteinases(MMP)-1,-2,-9,tissue inhibitor of metalloproteinase(TIMP)-1 and -2 were measured with reverse transcription-polymerase chain reaction(RT-PCR) test.Results After XGA administration with different doses and at different time points was adminstered,the gel electrophoresis product of RT-PCR in fibroblast culture system expressed the mRNA of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2;but the mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 decreased and the mRNA expression levels of MMP-1,-2,and -9 increased compared to those before XGA administration;the mRNA expression of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2 decreased or increased gradually with the increase of doses and the prolonged time of XGA use.The mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 were negatively related to the doses and action times of XGA(r=-0.927 to -0.637 P= 0 to 0.024);and the mRNA expression levels of MMP-1,-2 and -9 were positively related to the doses and action times of XGA(r=0.672 to 0.962 P=0 to 0.034).Conclusion XGA can markedly reduce the collagen formation of myocardial fibroblasts in rats,and its mechanisms are related to the inhibiting of collagen synthesis and the increase of collagen degradation.