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目的 定量检测转化生长因子 β1(TGF β1)和TGF β2在大鼠正常肝脏中肝细胞和汇管区间质细胞中的表达水平。方法 激光显微切割技术分离大鼠正常肝脏中的肝细胞和结缔组织细胞 ,实时荧光定量聚合酶链反应 (PCR)技术分析TGF β1和TGF β2的mRNA含量。 结果 激光显微切割技术成功地将肝脏结缔组织细胞从肝细胞中分离。TGF β1的mRNA含量在肝细胞中是TGF β2的 11倍左右 ,在肝脏汇管区间质细胞中接近TGF β2的 5倍左右。 结论 激光捕获显微切割技术结合实时荧光定量PCR技术能够针对性地精确分析不同的和极少量的细胞中基因表达 ,在肝脏中TGF β1的合成多于TGF β2。
Objective To quantitatively detect the expression of transforming growth factor β1 (TGF β1) and TGF β2 in hepatocytes and portal epithelial cells in normal rat liver. Methods Laser microdissection was used to separate the hepatocytes and connective tissue cells from normal liver of rats. The mRNA levels of TGF β1 and TGF β2 were analyzed by real-time fluorescence quantitative polymerase chain reaction (PCR). Results Laser microdissection successfully isolated hepatic connective tissue cells from hepatocytes. The mRNA level of TGF-β1 is about 11-fold that of TGF-β2 in hepatocytes and approximately 5-fold that of TGF-β2 in the hepatic portal cell compartments. Conclusion Laser capture microdissection combined with real-time fluorescence quantitative PCR can accurately and accurately analyze the gene expression in different and very few cells, and the synthesis of TGFβ1 in the liver is more than TGFβ2.