二甲双胍对鸡巨噬细胞HD11天然免疫相关基因表达影响及其抗禽白血病病毒能力分析

EFFECT OF METFORMIN ON THE EXPRESSION OF INNATE IMMUNERELATED GENES AND THE ABILITY OF ANTI-AVIAN LEUCOSIS VIRUS IN CHICKEN MACROPHAGES HD11

本研究旨在探讨二甲双胍(metformin)处理对鸡巨噬细胞HD11天然免疫相关基因表达及禽白血病病毒(Avian leukosisvirus,ALV)复制水平的影响。用2mmol/L二甲双胍处理鸡巨噬细胞HD11,处理48h后收集细胞,荧光定量PCR(RT-qPCR)检测天然免疫基因和内源性反转录病毒的表达。另外,将J亚群禽白血病病毒(J subgroup Avian leukosis virus,ALV-J)(MOI=5)接种于细胞板中,病毒感染细胞24 h后进行二甲双胍处理, 48 h后收集HD11细胞和上清分别进行RT-qPCR检测和ELISA(enzyme linked immunosorbent assay)检测。结果表明,二甲双胍显著上调HD11细胞天然免疫相关基因TLR3(P<0.01)、IFIH1(P<0.01)、IRF7(P<0.01)、STAT1(P<0.05)、IFN-α(P<0.01)、IFN-β(P<0.01)表达水平,并激活了鸡内源性反转录病毒的表达,抑制ALV-J复制水平。综上,二甲双胍可激活鸡巨噬细胞的天然免疫反应,从而提高鸡抗禽白血病病毒的能力。本研究在体外实验中验证二甲双胍具有抑制ALV增殖的作用,对家禽业的疾病防控有一定的应用价值。

The aim of the study was to explore the effect of metformin on the expression of the innate immune-related genes and avian leukosis virus replication levels in chicken macrophages HD11. The HD11 cells were treated with 2 mmol/L metformin. After 48 h,cells were collected and the expression of innate immune-related genes and endogenous retroviruses were measured by fluorescence quantitative PCR(RT-qPCR). In addition, HD11 cells were infected with J subgroup avian leukosis virus(ALV-J)(MOI = 5). The cells were treated with metformin for 24 hours and collected for RT-qPCR and supernatant was collected after 48 hours for ELISA testing. The results show that metformin significantly activated the expression of innate immune-related genes TLR3(P<0.01), IFIH1(P<0.01), IRF7(P<0.01), STAT1(P<0.05), IFN-α(P<0.01), IFN-β(P< 0.01) and endogenous retroviruses. The level of avian leukosis virus replication decreased significantly. Therefore, metformin activated the innate immune responses of the chicken macrophages and enhanced the antiviral ability.