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目的检测门静脉高压症(PHT)脾亢脾和正常脾巨噬细胞的差异表达基因,为从基因水平观察巨噬细胞在门静脉高压症脾亢发生中的作用奠定基础。方法提取门静脉高压症脾亢脾巨噬细胞和正常脾巨噬细胞的总RNA,分别用标有荧光素的dCTP反转录制备cDNA探针,将探针与含有14112点cDNA的Biostar-H140s cDNA表达谱芯片杂交后扫描荧光强度,上述方法重复3次,从而筛选出恒定的差异表达基因。结果3张芯片分别检测到896、1 330和898个差异表达基因。恒定的差异表达基因共有121个,占总基因数的0．86%。其中表达上调的已知基因有21个,表达下调的已知基因有73个。差异表达基因涉及离子通道和运输蛋白、细胞周期蛋白类、细胞骨架和运动、细胞受体、细胞信号和传递蛋白、代谢、免疫相关等多个方面。结论筛选出的差异表达基因,对了解脾脏巨噬细胞在门静脉高压症脾亢发生中的作用有重要意义。
Objective To detect differentially expressed genes of splenic hypersplenism and normal splenic macrophages in patients with portal hypertension (PHT), and lay a foundation for the observation of the role of macrophages in the pathogenesis of portal hypertrophy and hypersplenism. Methods The total RNA of splenic macrophages and splenic macrophages of splenic hypertrophy in portal hypertension was extracted. The cDNA probes were reverse transcribed with dCTP labeled with fluorescein, and the probes were ligated with Biostar-H140s cDNA containing 14112-point cDNA After the hybridization of the expression profiling chip, the fluorescence intensity was scanned, and the above method was repeated three times to screen for a constant differentially expressed gene. Results 896, 1 330 and 898 differentially expressed genes were detected by 3 chips respectively. A total of 121 differentially expressed genes, accounting for 0.86% of the total number of genes. There were 21 known genes whose expression was up-regulated, and 73 genes were down-regulated. Differentially expressed genes involved in ion channels and transport proteins, cyclin, cytoskeleton and movement, cell receptors, cell signaling and transfer protein, metabolism, immune related and many other aspects. Conclusion The differentially expressed genes screened out are of great significance for understanding the role of splenic macrophages in the pathogenesis of hypersplenism in patients with portal hypertension.