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目的探讨寡聚脱氧核苷酸(Cp G-ODN)体外对流感病毒复制的抑制作用。方法 Cp G-ODN刺激人喉癌上皮细胞(Hep-2),采用荧光定量PCR观察Toll样受体9(TLR9)及细胞因子的表达情况;Cp G-ODN体外预处理Hep-2细胞后感染流感病毒,采用荧光定量PCR法检测细胞内流感病毒拷贝数,了解不同剂量和不同时间Cp GODN预处理对Hep-2细胞中流感病毒复制影响。结果 Cp G-ODN处理72 h后,Hep-2细胞内TLR9 mRNA相对表达量(18.6±3.6)最高(P<0.05);细胞因子白细胞介素-6(IL-6)、干扰素-β(IFN-β)和肿瘤坏死因子-α(TNF-α)的mRNA表达高峰也出现在72 h,分别为(96.2±21.4)、(33.7±5.4)和(17.2±3.4)(均P<0.05)。各Cp GODN预处理组病毒拷贝数均低于对照组(P<0.05),并呈剂量和时间依赖关系,其中Cp G-ODN终浓度5μmol/L预处理Hep-2细胞24 h抑制流感病毒复制效果最明显(P<0.05)。结论 Cp G-ODN作为一种新型的免疫调节剂,可诱发机体免疫效应,抑制流感病毒复制。
Objective To investigate the inhibitory effect of CpG ODN on influenza virus replication in vitro. Methods Hep-2 cells were stimulated with CpG-ODN and the expression of TLR9 and cytokines were detected by real-time PCR. CpG-ODN was used to infect Hep-2 cells in vitro Influenza virus, Fluorescent quantitative PCR was used to detect the intracellular influenza virus copy number to understand the different doses and different time Cp GODN pretreatment of Hep-2 cells in the replication of influenza virus. Results The expression of TLR9 mRNA in Hep-2 cells was highest at 18 h (P <0.05) after CpG-ODN treatment for 72 h (P <0.05). The levels of IL-6 and IFN- The mRNA expression peak of IFN-β and TNF-α also appeared at 72 h (96.2 ± 21.4), (33.7 ± 5.4) and (17.2 ± 3.4), respectively (all P <0.05) . The CpGODN pretreatment group virus copy number were lower than the control group (P <0.05), and dose-dependent and time-dependent manner, CpG-ODN final concentration of 5μmol / L Pretreatment Hep-2 cells inhibit influenza virus replication The effect is most obvious (P <0.05). Conclusion Cp G-ODN, as a novel immunomodulator, can induce the immune response and inhibit the replication of influenza virus.