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摘要:目的比较原发性肾病综合征患儿和健康儿童外周血单个核细胞基因表达谱的差异,筛选原发性肾病综合征相关基因,绘制原发性肾病综合征的分子肖像,系统阐明原发性肾病综合征发生的分子机制。方法分离6例原发性肾病综合征患儿和6例与之匹配的健康儿童的外周血单个核细胞,用TRIzol一步法提取外周血单个核细胞的总RNA,再用逆转录-PCR法以Cy5和Cy3分别标记原发性肾病综合征患儿和健康儿童的mRNA而成为cDNA探针。将两种探针混合后与点有8096条基因的芯片杂交,经严格洗涤后用GenePix4000B扫描仪扫描杂交信号,用GenePixPro3.0软件分析扫描结果。结果8096条被检基因中共有418条在PNS患儿和健康儿童的PBMC中存在差异表达,其中128条基因在PNS患儿的PBMC中表达升高,290条基因表达降低,102条基因在本研究的6对PNS患儿和健康儿童配伍对照中均存在差异表达。这102条基因根据功能可分为10类:免疫相关基因、细胞骨架相关基因、信号转录相关基因、细胞周期及增殖凋亡相关基因、癌基因和抑癌基因、基因复制和表达相关基因、离子通道和转运蛋白相关基因、受体相关基因、代谢相关基因及其它功能尚不清楚的基因。结论原发性肾病综合征的发生、发展是许多相互作用的基因共同作用的结果。
Abstract: Objective To compare the gene expression profiles of peripheral blood mononuclear cells in children with primary nephrotic syndrome and healthy children, screen the related genes of primary nephrotic syndrome and draw the molecular portrait of primary nephrotic syndrome. Molecular mechanism of nephrotic syndrome. Methods Peripheral blood mononuclear cells from 6 children with primary nephrotic syndrome and 6 matched healthy children were isolated. The total RNA was extracted from peripheral blood mononuclear cells by one-step TRIzol method and then analyzed by reverse transcription-PCR Cy5 and Cy3 were used as cDNA probes to label the mRNA of children with primary nephrotic syndrome and healthy children, respectively. The two probes were mixed with 8096 gene chip hybridization point, after a rigorous washing with the GenePix4000B scanner scanning hybridization signal, using the GenePixPro3.0 software analysis of the scan results. Results A total of 418 out of 8096 test samples were differentially expressed in PBMCs of children with PNS and healthy children, of which 128 were increased in PBMCs, 290 were decreased, 102 were in this The study of 6 pairs of PNS children and healthy children compatibility control there are differences in expression. These 102 genes can be classified into 10 categories according to their function: immune related genes, cytoskeleton related genes, signal transduction related genes, cell cycle and proliferation related genes, oncogenes and tumor suppressor genes, gene replication and expression related genes, ions Channel and transporter-related genes, receptor-related genes, metabolic-related genes, and other genes whose function is unclear. Conclusions The occurrence and development of primary nephrotic syndrome are the result of the interaction of many interacting genes.