家族聚集性慢性HBV感染者外周血单个核细胞自噬相关基因表达谱研究

Expression of autophagy modulating genes in PBMCs from familial clustering patients with chronic virus hepatitis B

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 Evidences show that the autophagy is involved in the chronic hepatitis B infection. In order to address this issue from a molecular standpoint, expression of an array of gene coding for key autophagy associated genes in peripheral mononu- clear cells were measured and analyzed. Methods cRNA prepared from PBMCs in a family of clustering HBV infection including 11 CHB patients and 9 healthy spouses was hybridized to high-density oligouncleotide arrays （HG-U133A 2.0 Human Gene- Chips, Affymetrix）, which interrogate the expression of ^-22 000 human ESTs. Primary image obtained from scanner was analyzed with DNT software package. Results 19 autophagy-modulating genes out of 22 000 ESTs were identified differently. Among the 19 genes, 12 displayed increased expression and 7 genes decreased expression in CHB compared with those in healthy control. Functional analyses show that those genes were closely involved in the initiation, elongation, sequestration, infusion and degrada- tion. Conclusion A differential expression of genes related autophagy between patients with chronic hepatitis B and healthy indi- vidual were indented, a decreased autophagy in PBMCs was observed, which may be contributed to lymphocyte senescence.