Divison of Molecular Virology, National Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore, Pakistan.
Hazara University, Mansehra, Pakistan.
Arch Virol. 2020 Apr;165(4):809-822. doi: 10.1007/s00705-020-04564-z. Epub 2020 Feb 27.
Oxidative stress is the process by which reactive molecules and free radicals are formed in cells. In this study, we report the blood-based gene expression profile of oxidative stress and antioxidant genes for identifying surrogate markers of liver tissue in chronic hepatitis C (CHC) patients by using real-time PCR. A total of 144 untreated patients diagnosed with CHC having genotype 3a and 20 healthy controls were selected for the present study. Liver biopsy staging and grading of CHC patients were performed using the METAVIR score. Total RNA was extracted from liver tissue and blood samples, followed by cDNA synthesis and real-time PCR. The relative expression of genes was calculated using the ΔΔCt method. The expression profile of 84 genes associated with oxidative stress and antioxidants was determined in liver tissue and blood samples. In liver tissue, 46 differentially expressed genes (upregulated, 27; downregulated, 19) were identified in CHC patients compared to normal samples. In blood, 61 genes (upregulated, 51; downregulated; 10) were significantly expressed in CHC patients. A comparison of gene expression in liver and whole blood showed that 20 genes were expressed in a similar manner in the liver and blood. The expression levels of commonly expressed liver and blood-based genes were also correlated with clinical factors in CHC patients. A receiver operating curve (ROC) analysis of oxidative stress genes (ALB, CAT, DHCR24, GPX7, PRDX5, and MBL2) showed that infections in patients with CHC can be distinguished from healthy controls. In conclusion, blood-based gene expression can reflect the behavior of oxidative stress genes in liver tissue, and this blood-based gene expression study in CHC patients explores new blood-based non-invasive biomarkers that represent liver damage.
氧化应激是细胞内形成反应性分子和自由基的过程。在这项研究中,我们通过实时 PCR 报告了氧化应激和抗氧化基因的基于血液的基因表达谱,以鉴定慢性丙型肝炎(CHC)患者肝组织的替代标志物。共选择了 144 名未经治疗的诊断为 CHC 的基因型 3a 患者和 20 名健康对照者进行本研究。使用 METAVIR 评分对 CHC 患者进行肝活检分期和分级。从肝组织和血液样本中提取总 RNA,然后进行 cDNA 合成和实时 PCR。使用 ΔΔCt 法计算基因的相对表达。确定了与氧化应激和抗氧化剂相关的 84 个基因在肝组织和血液样本中的表达谱。在肝组织中,与正常样本相比,CHC 患者中有 46 个差异表达基因(上调 27 个;下调 19 个)。在血液中,CHC 患者中有 61 个基因(上调 51 个;下调 10 个)显著表达。肝和全血中基因表达的比较表明,20 个基因在肝和血液中以相似的方式表达。常用的肝和基于血液的基因的表达水平也与 CHC 患者的临床因素相关。氧化应激基因(ALB、CAT、DHCR24、GPX7、PRDX5 和 MBL2)的接收器工作曲线(ROC)分析表明,CHC 患者的感染可以与健康对照者区分开来。总之,基于血液的基因表达可以反映肝组织中氧化应激基因的行为,这项针对 CHC 患者的基于血液的基因表达研究探索了新的基于血液的非侵入性生物标志物,代表了肝损伤。