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SOCS1 和 SOCS3 基因在干扰素治疗和直接作用抗病毒药物治疗丙型肝炎患者中的表达。

Expression of SOCS1 and SOCS3 Genes in Interferon-Treated and Direct-Acting Antiviral Drugs-Treated Hepatitis C Patients.

机构信息

1 Molecular Virology Laboratory Centre for Applied Molecular Biology (CAMB), University of the Punjab , Lahore, Pakistan .

2 Divison of Molecular Virology, Center of Excellence in Molecular Biology (CEMB), University of the Punjab , Lahore, Pakistan .

出版信息

J Interferon Cytokine Res. 2018 Jun;38(6):255-260. doi: 10.1089/jir.2017.0138.

Abstract

Genetics of host plays a significant role in susceptibility and pathogenesis of disease. During hepatitis C virus (HCV) infection, HCV proteins interfere with interferon (IFN) signaling pathways and upregulate transcription of suppressor of cytokine signaling 1 and 3 genes (SOCS1 and SOCS3), which results in impaired immune response. In this study, we evaluated relative expression of SOCS1 and SOCS3 in untreated HCV patients and patients treated with 2 different treatment strategies that are, (IFN therapy and direct-acting antiviral (DAA) drug regimen. To study gene expression, peripheral blood mononuclear cells (PBMCs) were isolated by using Histopaque. Total RNA was extracted from PBMCs by using BIOzol. Nine microgram of total RNA from each sample was used and reverse transcribed into single-stranded complementary DNA (cDNA) by using M-MLV reverse transcriptase (Invitrogen). The synthesized cDNA was diluted to a final concentration of 500 ng/μL. This diluted cDNA was further used for expression analysis of SOCS1and SOCS3 genes using Rotor Gene Q Real-Time PCR Detection System (QIAGEN). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was amplified as a housekeeping gene. We found that the SOCS1 expression in IFN and DAA-treated patient groups was 5.4 fold and 1.2 fold, respectively, high compared with the healthy controls (IFN versus healthy, P = 0.019 and DAA versus healthy, P = 0.91), whereas the SOCS3 expression in IFN and DAA-treated patient groups was 3.7 fold and 2 fold, respectively, high in comparison with the expression in healthy controls (IFN versus healthy, P = 0.025 and DAA versus healthy, P = 0.03). We also found a significant difference in the relative expression of SOCS1 and SOCS3 in DAAs-treated and IFN/ribavirin (RBV)-treated and untreated individual. We concluded that by targeting HCV proteins with DAAs, SOCS1, and SOCS3 transcription can be more effectively normalized compared to the treatment with IFN/RBV therapy.

摘要

宿主遗传学在疾病的易感性和发病机制中起着重要作用。在丙型肝炎病毒 (HCV) 感染期间,HCV 蛋白干扰干扰素 (IFN) 信号通路,并上调细胞因子信号转导抑制因子 1 和 3 基因 (SOCS1 和 SOCS3) 的转录,导致免疫反应受损。在这项研究中,我们评估了未经治疗的 HCV 患者和接受两种不同治疗策略的患者(IFN 治疗和直接作用抗病毒 (DAA) 药物治疗)中 SOCS1 和 SOCS3 的相对表达。为了研究基因表达,使用 Histopaque 分离外周血单核细胞 (PBMC)。使用 BIOzol 从 PBMC 中提取总 RNA。从每个样本中提取 9μg 的总 RNA,并使用 M-MLV 逆转录酶 (Invitrogen) 将其逆转录为单链互补 DNA (cDNA)。合成的 cDNA 稀释至终浓度为 500ng/μL。进一步使用 Rotor Gene Q 实时 PCR 检测系统 (QIAGEN) 对 SOCS1 和 SOCS3 基因的表达进行分析。甘油醛-3-磷酸脱氢酶 (GAPDH) 被扩增为管家基因。我们发现,与健康对照组相比,IFN 和 DAA 治疗组的 SOCS1 表达分别高 5.4 倍和 1.2 倍(IFN 与健康对照组,P=0.019;DAA 与健康对照组,P=0.91),而 SOCS3 在 IFN 和 DAA 治疗组的表达分别比健康对照组高 3.7 倍和 2 倍(IFN 与健康对照组,P=0.025;DAA 与健康对照组,P=0.03)。我们还发现,在 DAA 治疗组、IFN/利巴韦林 (RBV) 治疗组和未治疗组中,SOCS1 和 SOCS3 的相对表达存在显著差异。我们得出的结论是,通过用 DAA 靶向 HCV 蛋白,可以更有效地使 SOCS1 和 SOCS3 的转录正常化,而不是用 IFN/RBV 治疗。

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