芪莲茅根汤含药血清对大鼠肾小球系膜细胞凋亡相关基因表达谱的影响  

Effects of Qilian-maogen Tang medicated serum on apoptosis related gene expression profile of rats＇ mesangial Cells

作　　者：刘建华[1] 曾金雄[1] 兰凤华[1] 杨家辉[1] 

机构地区：[1]南京军区福州总医院中医专科中心,福州350025

出　　处：《国际中医中药杂志》2007年第3期131-136,共6页International Journal of Traditional Chinese Medicine

摘　　要：目的利用cDNA基因表达谱芯片观察芪茅莲根汤含药血清干预后的大鼠肾小球系膜细胞基因表达水平的变化，通过诱导肾小球系膜细胞凋亡相关基因的表达谱变化，探讨其在凋亡信号转导途径中的机制。方法建立大鼠肾小球系膜细胞培养体系，用Trizol一步法、抽提法分别从正常对照组、含药血清组的大鼠肾小球系膜细胞中抽提mRNA，经逆转录后合成荧光分子（Cy3／Cy5）标记的cDNA探针，与含有11260个cDNA基因的大鼠12K基因表达谱芯片杂交，将图像导入分析软件Imagene进行图像分析，数据导入分析软件Genespring进行标准化处理，计算ratio值（两种荧光Cy3、Cy5的比值）。分析正常对照组、含药血清组之间差异表达的基因，对所获得的基因进行生物信息学分析。结果含药血清组与正常对照组中差异表达上调的基因147条、下调的168条；涉及参与细胞凋亡发生以及能量代谢、蛋白合成、信号转导和激活因子。与细胞凋亡相关的差异表达基因37条，占芯片基因总数的0．3286％，其中表达上调的21条（平均Ratio值为3．975），下调的16条（平均Ratio值为0．402）；生物信息学分析表明，Bcl-2、Bak1、Caspase9、Timp1、Cdk5、Pdgfra、Igfbp5、Irf1等在芪莲茅根汤含药血清诱导大鼠肾脏系膜细胞凋亡的调控中发挥了重要作用。结论芪茅莲根汤含药血清诱导肾小球系膜细胞调亡涉及多个基因，通过细胞内、外信号传导途径共同调节完成，从而调控肾小球系膜细胞的增殖。Objective To study the effects of Qilian-maogen Tang medicated serum on the gene expression profile of rat＇s mesangial cells, and to reveal the possible mechanism of signal transduction on apoptosis. Methods The mesangial cells were cultured in Qilian-maogen Tang medicated serum . Total RNA were extracted by using TRIzol from rat＇s mesangial cells both treated and untreated by Qilian-maogen Tang. Two cDNA probes were made via reverse transcription, which were labeled by Cy3-dUTP or Cy5-dUTP fluorescence dyes respectively. A microarrary chip contained 11260 rat 12K cDNA was used for hybridization. The analyses of differential expression profiles were performed by Genespring software, and then subjected to standardization and ratio analysis. Results In this study, 147 up-regulated genes and 168 down-regulated genes were detected respectively, and 37 apoptosis related genes were found among the total genes in cDNA microarray （0.3286%）. Within the 37 apoptosis related genes, 21 genes were up-regulated （average Ratio： 3.975） and 16 genes were down-regulated （average Ratio： 0.402）. Furthermore, it was also found by bioinformatics analyses that some genes within the 37 genes, such as Bc1-2、 BAK1, Caspase9, TIMP1, Cdk5, Pdgfra, Igfbp5 and Irfl should play critical roles in modulation of apoptosis of mesangial cell when induced by Qilian-maogen Tang medicated serum in vitro. Conclusion The results demonstrated that the mechanisms of apoptosis of meseangial cells induced by Qilian-maogen Tang medicated serum should be likely co-regulated by a variety of different genes, meanwhile, its signal transduction pathways are modulated through both intra and extra cellular process. The study gives a further understanding to the mechanism of the Qilian-maogen Tang in improving the kidney function of IgAN rat.

关 键 词：芪莲茅根汤 含药血清 肾小球系膜 凋亡 增殖 基因芯片 

分 类 号：R28[医药卫生—中药学]