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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-2mRNA表达;与予XGA前比较,Ⅰ、Ⅲ、Ⅳ型胶原,TIMP-1、-2mRNA表达水平有所下降,而MMP-1、-2、-9mRNA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、-2mRNA表达水平与XGA的剂量和作用时间呈负相关(r=-0.927~-0.637P=0~0.024);MMP-1、-2、-9mRNA表达水平与XGA剂量和作用时间呈正相关(r=0.672~0.962P=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。
Objective To investigate the effect of astragaloside IV (XGA) on the collagen of rat cardiac fibroblasts and its dose-effect and time-effect relationship. Methods FBC cultured in rat cardiac fibroblasts (FBC) was obtained by trypsin digestion method. FBC culture system was established. FBC culture system was added with different concentrations of XGA and different reaction time. After RNA extraction, reverse transcription PCR was used. RT-PCR assay was used to detect the expression levels of type I, III, IV collagenase, matrix metalloproteinase (MMP-1, -2, -9) and its inhibitor (TIMP-1, -2) mRNA. After different time of action XGA, RT-PCR product gel electrophoresis of FBC culture system showed the expression of type I, III, IV collagen, MMP-1, -2, -9, TIMP-1 and -2 mRNA; compared with pre-XGA, Collagen types I, III, IV, and TIMP-1, -2 mRNA levels decreased, while MMP-1, -2, -9 mRNA levels increased, and gradually decreased with increasing XGA dose or duration of action. The levels of collagen type I, III, IV, and TIMP-1, -2 mRNA expression were negatively correlated with dose and duration of action of XGA (r = -0.927 - -0.637P = 0.024), and MMP-1, -2, The expression level of -9 mRNA was positively correlated with XGA dose and duration of action (r=0.672-0.962P=0-0.034).Conclusion XGA can reduce collagen formation in cardiac fibroblasts, and its mechanism may be inhibition of collagen. Synthesis, increased collagen degradation.