乙型肝炎病毒对HepG2细胞IL-17R信号通路的影响

目的:研究乙型肝炎病毒(hepatitis B virus,HBV)重组腺病毒对HepG2细胞的IL-17R和接头蛋白Act1表达的影响,以及HBV对IL-17诱导NF-B活化的影响.方法:采用实时荧光定量PCR(real-time PCR)检测HepG2细胞的IL-17、IL-17R和Act1的mRNA表达;蛋白免疫印迹法(Western blot)检测IL-17R和Act1的蛋白表达;免疫荧光检测NF-B核移位;ELISA检测上清的IL-17含量.结果:各组HepG2细胞培养上清液中均未检测到IL-17且亦未检测HepG2细胞有IL-17的mRNA表达;HBV重组腺病毒组的IL-17R mRNA和蛋白的表达明显低于相应浓度对照组(0.68±0.02vs0.89±0.03,0.33±0.06vs0.81±0.01,0.12±0.01vs0.86±0.05,P<0.05;蛋白:0.84±0.12vs1.01±0.13,0.56±0.09vs1.01±0.08,0.24±0.08vs0.98±0.05),且呈剂量和时间依赖性.但HBV重组腺病毒组与对照组比较,对HepG2细胞接头蛋白Act1的mRNA和蛋白表达水平无明显影响;同时HBV重组腺病毒能抑制IL-17R诱导Hep G2细胞的NF-B活化.但HBV重组腺病毒与对照组比较,对接头蛋白Act1在mRNA和蛋白表达水平上影响无明显变化;同时HBV重组腺病毒能抑制IL-17R诱导Hep G2细胞的NF-B活化.结论:HBV重组腺病毒可降低HepG2细胞的IL-17R mRNA和蛋白的表达,抑制IL-17R诱导Hep G2细胞的NF-B活化,对HepG2细胞的IL-17R信号通路发挥抑制作用.

Influence of HBV on the interleukin-17 receptor signaling pathway in HepG2 cells

AIM: To investigate whether hepatitis B virus influences the expression of interleukin-17 (IL-17) receptor and the adaptor Act1 in HepG2 cells. METHODS: The mRNA and protein expression of IL-17 receptor and the adaptor Act1 was determined by real-time PCR and Western blot, respectively. NF-kappa B nuclear translocation was detected by immunofluorescence. The content of IL-17 in cell supernatants was measured by ELISA. RESULTS: IL-17 was not detected in the culture supernatants of HepG2 cells, and IL-17 mRNA expression was not detected in HepG2 cells. The mRNA and protein expression of IL-17 receptor was significantly lower in HepG2 cells infected with HBV recombinant adenovirus than in cells infected with corresponding concentration of wild adenovirus (mRNA: 0.68±0.02 0.89±0.03, 0.33±0.06 vs 0.81±0.01, 0.12±0.01 vs 0.86±0.05, all P < 0.05; protein: 0.84±0.12 vs 1.01±0.13, 0.56± 0.09 vs 1.01±0.08, 0.24±0.08 vs 0.98±0.05, all P < 0.05), and the impact of HBV on the IL-17 receptor expression was dose-and time-dependent. However, HBV had no significant impact on the expression of adaptor Act1. Furthermore, HBV recombinant adenovirus inhibited NF-B activation induced by IL-17 receptor. CONCLUSION: Our results indicate that HBV recombinant adenovirus can down-regulate the mRNA and protein expression of IL-17 receptor, inhibit NF-B activation induced by IL-17R, and thereby prevent the IL-17 receptor signaling pathway in HepG2 cells.