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目的应用寡核苷酸基因芯片技术,研究家族聚集性慢性HBV感染者外周血单个核细胞(PBMCs)自噬相关基因的表达谱,并进行功能分析。方法:在聚集性HBV感染家族中,选取患者9例和对照11例,提取PBMCs中的RNA,与涵盖2.2万个EST的寡核苷酸表达谱基因芯片U133A 2.0杂交,通过Affymetrix扫描仪和DNT分析软件比较患病组与对照组PBMC免疫相关基因的的表达谱,获得基因的相对表达比值;对差异表达基因进行功能分析。结果:在2.2万个EST中,192个与自噬相关,初筛出19个自噬相关差异表达基因,7个基因表达上调,17个表达下调,差异表达基因主要与自噬的起始、延伸、包裹和降解过程有关;功能分析发现慢性HBV感染者PBMCs的自噬水平下降。结论:慢性HBV感染者的PBMCs的自噬基因存在差异表达,其噬水平下降,可能与淋巴细胞老化过程有关。
Objective To study the expression profiles of autophagy-related genes in peripheral blood mononuclear cells (PBMCs) from familial chronic hepatitis B patients with chronic hepatitis B virus infection by oligonucleotide microarray. Methods: Nine patients and 11 controls were recruited in the cluster of HBV infection. RNA was extracted from PBMCs and hybridized with U133A 2.0, an oligonucleotide microarray DNA microarray covering 22,000 ESTs. Affymetrix scanners and DNT Analysis software was used to compare the expression profiles of PBMC immune-related genes in the diseased group and the control group, and the relative expression ratio of the genes was obtained. The differentially expressed genes were analyzed. Results: Of the 22,000 ESTs, 192 were related to autophagy. Nineteen autophagy-related differentially expressed genes were initially screened, seven were up-regulated and 17 were down-regulated. The differentially expressed genes were mainly associated with the initiation of autophagy, Extension, parcel and degradation process; functional analysis found that chronic HBV infected PBMCs decreased autophagy levels. CONCLUSION: Autophagy genes of PBMCs from patients with chronic HBV infection are differentially expressed and their levels are decreased, which may be related to the aging process of lymphocytes.