摘要
目的:探索TNF-α抑制剂对丙型肝炎病毒感染肝细胞的影响。方法:体外培养肝癌细胞株Huh7细胞,分为未处理组、NFκB抑制剂[6-amino-4-(4-phenoxyphenylethylamino)quinazoline,QNZ]处理组、咖啡酸苯乙酯(caffeic acid phenethyl ester,CAPE)处理组和TNF-α抑制剂来那度胺(Lenalidomide)处理组,每组设置浓度梯度为100μmol/L至1.28 nmol/L,5倍倍比稀释,通过CCK8法检测各抑制剂对细胞增殖的影响;Huh7细胞经抑制剂处理后,再以丙型肝炎病毒感染,通过荧光定量PCR和免疫荧光法检测细胞内病毒的感染水平。结果:100μmol/L QNZ处理组(0.730±0.056)和100μmol/L CAPE处理组(1.040±0.014)细胞在450 nm处的吸光度(absorbance,A)值均低于未处理组(1.190±0.040,P=0.000)。Lenalidomide各浓度处理组(100μmol/L处理组,1.250±0.016)细胞增殖水平与未处理组相当(P=0.306)。在抗病毒活性方面,100 nmol/L Lenalidomide处理组[(3.79±0.88)×105]和QNZ处理组[(6.64±1.11)×105]HCV RNA水平明显低于未处理组[(1.51±0.45)×10~6,P=0.000],而CAPE处理组[(1.61±0.15)×10~6]与未处理组HCV RNA水平相当(P=0.383)。结论:TNF-α抑制剂Lenalidomide对细胞毒性较小,能够在细胞水平上抑制丙型肝炎病毒的感染。
Objective：To research the effect of TNF-α inhibitors on hepatitis C virus（HCV） infection in hepatocytes. Methods：Hep- atoma Huh7 cells were cultured in vitro. Huh7 cells divided into four groups：untreated group,NFKB inhibitors [6-amino-4-（4-phe- noxyphenylethylamino） quinazoline, QNZ] treated group,caffeic acid phenethyl ester （CAPE）） treated group and TNF-α inhibitors （Lenalidomide） treated group. Concentrations of the inhibitors were from 100 μmol/L to 1.28 nmol/L with 5-fold serial dilution in each group. Cell proliferation was observed via CCK8 methods. Huh7 cells pre-treated with inhibitors were followed with HCV infection. The intracellular HCV RNA and protein levels were determined by quantitative PCR and immunofluorescence assay. Results：The ab- sorbance values at 450 nm of 100 μmol/L QNZ treatment group（0.730 ± 0.056） and 100 μmol/L CAPE treatment group（1.040 ± 0.014） were lower than those of untreated group（1.190 ± 0.040,P=0.000）. The proliferation level of each concentration of Lenalido- mide treatment group（100 μmol/L treatment group, 1.250± 0.016） was similar with untreated group（P=0.306）. In terms of antiviral activity,the HCV RNA levels of 100 nmol/L Lenalidomide treatment group[（3.79 ± 0.88） × 10^5] and QNZ treatment group[（6.64 ± 1.11 ） ×10^5] were significantly lower than those of untreated group[（ 1.51± 0.45） × 10^6, P=0.000]. CAPE treatment group[（ 1.61 ± 0.15） ×10^6] had a similar HCV RNA level with untreated group（P=0.383）. Conclusion ：TNF-α inhibitor Lenalidomide is less toxic to cells and can inhibit hepatitis C virus infection in hepatocytes.
出处
《重庆医科大学学报》
CSCD
北大核心
2017年第11期1488-1493,共6页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81301445)
院管课题基金资助项目(编号:2013YG-B055)
