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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 explore the effect of TNF-α inhibitor on hepatitis C virus-infected hepatocytes. Methods: The hepatocellular carcinoma cell line Huh7 was cultured in vitro and divided into untreated group, 6-amino-4- (4-phenoxyphenylethylamino) quinazoline, QNZ treatment group, caffeic acid phenethyl ester ) Treatment group and Lenalidomide treatment group, the concentration gradient was set at 100μmol / L to 1.28nmol / L in each group and diluted by 5 times. CCK8 assay was used to detect the effect of each inhibitor on cell proliferation Huh7 cells were treated with inhibitors and then infected with Hepatitis C virus. Fluorescence quantitative PCR and immunofluorescence were used to detect the intracellular viral infection. RESULTS: The absorbance (A) values at 450 nm in 100 μmol / L QNZ and 1.040 ± 0.014 cells treated with 100 μmol / L CAPE were significantly lower than those in untreated (1.190 ± 0.040, P = 0.000). Lenalidomide concentration of the treatment group (100μmol / L treatment group, 1.250 ± 0.016) cell proliferation levels compared with the untreated group (P = 0.306). In the antiviral activity, the level of HCV RNA in 100 nmol / L Lenalidomide treated group [(3.79 ± 0.88) × 105] and QNZ treated group [(6.64 ± 1.11) × 105] was significantly lower than that in untreated group [(1.51 ± 0.45) × 10 ~ 6, P = 0.000], while CAPE treatment group [(1.61 ± 0.15) × 10 ~ 6] had the same level of HCV RNA as untreated group (P = 0.383). Conclusion: Lenalidomide, a TNF-αinhibitor, is less toxic to cells and can inhibit the infection of hepatitis C virus at the cellular level.