Lysosomotropic agents as HCV entry inhibitors
Lysosomotropic agents as HCV entry inhibitors
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DOI：
10.1186/1743-422x-8-163
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发表时间：
2011-04-12
期刊：
影响因子：
4.8
通讯作者：
Riazuddin, Sheikh
中科院分区：
文献类型：
--
作者：
Ashfaq, Usman A.;Javed, Tariq;Riazuddin, Sheikh
HCV有两种包膜蛋白E1和E2，它们通过两种主要途径进入细胞：在质膜上的直接融合和受体介导的内吞作用。HCV包膜蛋白的融合由内体内的低pH触发。Lysosomotropic agents（LA）如氯喹和氯化铵（NH4Cl）是弱碱，以质子化形式渗透溶酶体并增加细胞内pH。为了研究LA（氯喹和NH4Cl）对pH依赖性内吞作用的抗病毒作用，制备了1a和3a基因型的HCV假颗粒（HCVpp）并用于感染肝细胞。采用MTT细胞增殖实验检测氯喹和氯化铵对肝细胞的毒性作用。为了进行氯喹和NH4Cl的抗病毒筛选，在存在或不存在不同浓度的氯喹和NH4Cl的情况下，用3a和1a基因型的HCVpp感染肝细胞，并通过使用发光计测定荧光素酶活性。结果表明，氯喹和NH4Cl分别在50 μ M和10 mM浓度下显示出超过50%的病毒感染性降低。这些结果表明，通过增加溶酶体pH值在融合步骤抑制HCV将是治疗慢性HCV的更好选择。
HCV has two envelop proteins named as E1 and E2 which play an important role in cell entry through two main pathways: direct fusion at the plasma membrane and receptor-mediated endocytosis. Fusion of the HCV envelope proteins is triggered by low pH within the endosome. Lysosomotropic agents (LA) such as Chloroquine and Ammonium chloride (NH4Cl) are the weak bases and penetrate in lysosome as protonated form and increase the intracellular pH. To investigate the antiviral effect of LA (Chloroquine and NH4Cl) on pH dependent endocytosis, HCV pseudoparticles (HCVpp) of 1a and 3a genotype were produced and used to infect liver cells. The toxicological effects of Chloroquine and NH4Cl were tested in liver cells through MTT cell proliferation assay. For antiviral screening of Chloroquine and NH4Cl, liver cells were infected with HCVpp of 3a and 1a genotype in the presence or absence of different concentrations of Chloroquine and NH4Cl and there luciferase activity was determined by using a luminometer. The results demonstrated that Chloroquine and NH4Cl showed more than 50% reduction of virus infectivity at 50 mu M and 10 mM concentrations respectively. These results suggest that inhibition of HCV at fusion step by increasing the lysosomal pH will be better option to treat chronic HCV.