Department of Environmental Sciences, COMSATS Institute of Information Technology, Abbottabad, Pakistan.
Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Biomed Pharmacother. 2016 Oct;83:881-891. doi: 10.1016/j.biopha.2016.08.002. Epub 2016 Aug 9.
Discovery of alternative and complementary regimens for HCV infection treatment is a need of time from clinical as well as economical point of views. Low cost of bioactive natural compounds production, high biochemical diversity and inexistent/milder side effects contribute to new therapies. Aim of this study is to clarify anti-HCV role of Taraxacum officinale, a natural habitat plant rich of flavonoids. In this study, methanol extract of T. officinale leaves was initially analyzed for its cytotoxic activity in human hepatoma (Huh-7) and CHO cell lines. Hepatoma cells were transfected with pCR3.1/Flagtag/HCV NS5B gene cloned vector (genotype 1a) along with T. officinale extract. Considering NS5B polymerase as potential therapeutic drug target, twelve phytochemicals of T. officinale were selected as ligands for molecular interaction with NS5B protein using Molecular Operating Environment (MOE) software. Sofosbuvir (Sovaldi: brand name) currently approved as new anti-HCV drug, was used as standard in current study for comparative analysis in computational docking screening. HCV NS5B polymerase as name indicates plays key role in viral genome replication. On the basis of which NS5B gene is targeted for determining antiviral role of T. officinale extract and 65% inhibition of NS5B expression was documented at nontoxic dose concentration (200μg/ml) using Real-time PCR. In addition, 57% inhibition of HCV replication was recorded when incubating Huh-7 cells with high titer serum of HCV infected patients along with leaves extract. Phytochemicals for instance d-glucopyranoside (-31.212 Kcal/mol), Quercetin (-29.222 Kcal/mol), Luteolin (-26.941 Kcal/mol) and some others displayed least binding energies as compared to standard drug Sofosbuvir (-21.0746 Kcal/mol). Results of our study strongly revealed that T. officinale leaves extract potentially blocked the viral replication and NS5B gene expression without posing any toxic effect on normal fibroblast cells of body.
从临床和经济角度来看,发现替代和补充 HCV 感染治疗方案是当务之急。生物活性天然化合物生产成本低、生化多样性高且副作用轻微,有助于开发新疗法。本研究旨在阐明蒲公英(一种富含类黄酮的天然栖息地植物)的抗 HCV 作用。在这项研究中,最初分析了蒲公英叶甲醇提取物对人肝癌(Huh-7)和 CHO 细胞系的细胞毒性作用。肝癌细胞与 pCR3.1/Flagtag/HCV NS5B 基因克隆载体(基因型 1a)共转染,并加入蒲公英提取物。鉴于 NS5B 聚合酶作为潜在的治疗药物靶点,使用分子操作环境(MOE)软件选择了蒲公英的 12 种植物化学物质作为与 NS5B 蛋白相互作用的配体。目前已批准索非布韦(Sovaldi:商标名)作为新型抗 HCV 药物,在本研究中用作比较分析计算对接筛选的标准。HCV NS5B 聚合酶顾名思义在病毒基因组复制中发挥关键作用。基于此,针对 NS5B 基因进行靶向,以确定蒲公英提取物的抗病毒作用,在非毒性剂量浓度(200μg/ml)下,使用实时 PCR 记录到 NS5B 表达抑制了 65%。此外,当用 HCV 感染患者的高滴度血清孵育 Huh-7 细胞时,同时加入叶提取物,记录到 HCV 复制抑制了 57%。类黄酮,例如 D-吡喃葡萄糖苷(-31.212 Kcal/mol)、槲皮素(-29.222 Kcal/mol)、木犀草素(-26.941 Kcal/mol)和其他一些物质与标准药物索非布韦(-21.0746 Kcal/mol)相比,显示出最低的结合能。我们的研究结果强烈表明,蒲公英叶提取物在不对身体正常成纤维细胞造成任何毒性作用的情况下,潜在地阻止了病毒复制和 NS5B 基因表达。