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目的:应用基因二代测序技术检测肝星状细胞(hepatic stellate cell,HSC)在压力作用后基因表达谱改变及可能机制.方法:分离培养原代肝星状细胞,培养14 d加载10 mmHg的压力1 h,提取总RNA,逆转录cDNA,用DEG-Seq对实验组与对照组进行比较,筛选得到差异基因.结果:肝星状细胞中979条基因出现差异表达,其中14条基因表达差异最为显著,10条基因表达上调,4条基因表达下调,这些基因功能与合成代谢、免疫应答、细胞凋亡等相关.结论:应用基因二代测序技术筛选出压力作用下肝星状细胞差异表达的基因,其可能通过调控这些基因来适应外界压力刺激.
OBJECTIVE: To detect the changes of gene expression profiles of hepatic stellate cells (HSC) under pressure and to explore the possible mechanisms by gene sequencing.Methods: Primary hepatic stellate cells (HSCs) were isolated and cultured and cultured for 10 days Pressure for 1 h, total RNA was extracted, cDNA was reverse transcribed, and DEG-Seq was used to compare the experimental group with the control group.Results: 979 genes were differentially expressed in hepatic stellate cells, of which 14 genes were different The expression of 10 genes was up-regulated and the expression of 4 genes was down-regulated.The functions of these genes were associated with anabolism, immune response, apoptosis and so on.Conclusion: Gene expression of differentially expressed hepatic stellate cells Of genes that may adapt to external stress stimuli by regulating these genes.