[Notch信号通路与Toll样受体4在慢性丙型肝炎患者CD14+单核细胞功能上的相互作用]
[Interaction of notch signaling pathway with toll-like receptor 4 on the function of CD14+ monocytes in chronic hepatitis C patients].
作者信息
Li Y, Li L X, Zhang X J, Yuan L J, Xi F Y, Zhang H, Zhang L X
机构信息
Department of Infectious Diseases, Shaanxi Provincial People's Hospital and The Affiliated Hospital of Xi'an Medical University, Xi'an 710068, China.
Department of Gynecology, Shaanxi Provincial Hospital People's Hospital and The Affiliated Hospital of Xi'an Medical University, Xi'an 710068, China.
出版信息
Zhonghua Gan Zang Bing Za Zhi. 2019 Jul 20;27(7):527-532. doi: 10.3760/cma.j.issn.1007-3418.2019.07.010.
To observe the expressional changes in Notch signaling pathway and toll-like receptor 4 (TLR4) and their interactions on the functions of CD14(+) monocytes in chronic hepatitis C patients. A total of 24 treatment-naïve chronic hepatitis C cases and 10 healthy individuals, who visited Shaanxi Provincial People's Hospital from August to October 2017, were enrolled. Selected CD14(+) monocytes were stimulated by the Notch signaling pathway inhibitor DAPT or transfected with TLR4 siRNA, and the levels of Notch1, Notch2, Hes1 and Hes5 mRNA were detected by real-time quantitative PCR. TLR4 protein levels and phosphorylation of NF-κB was detected by Western blot. ELISA was used to detect the level of cytokines secreted from CD14(+) monocytes. A t-test or paired t-test was used for comparison between groups. The relative expression of Notch1 mRNA (3.97 ± 2.03 vs. 0.91 ± 0.76, < 0.01) and downstream of Notch signaling pathway (5.96 ± 2.31 vs. 0.99 ± 0.45, < 0.01), Hes1 mRNA and Hes5 mRNA (4.31 ± 1.05 vs. 0.84 ± 0.20, < 0.01) in CD14+ monocytes of chronic hepatitis C patients was significantly higher than that of healthy individuals. The relative expression of TLR4 mRNA (5.14 ± 1.09 vs. 1.27 ± 0.39) and protein level in CD14(+) monocytes of chronic hepatitis C patients were significantly higher than those of healthy individuals ( < 0.01). An inhibition of Notch signaling pathway with DAPT had reduced the relative expression level of TLR4 mRNA (2.58 ± 1.36 vs. 4.34 ± 1.88, < 0.05), protein expression and phosphorylation of NF-B in CD14(+) monocytes of chronic hepatitis C patients. Furthermore, the secretion level of MCP-1 [(94.32 ± 23.59) pg/ml vs. (64.07 ± 9.39) pg/ml, < 0.01] and IL-8 [(12.54 ± 4.89) pg/ml vs. (7.92 ± 3.01) pg/ml, < 0.05] was significantly reduced. TLR4 siRNA transfection reduced the expressions of Notch1 mRNA (2.09 ± 1.72 vs. 3.73 ± 1.75, < 0.05), Hes1 (2.87 ± 0.84 vs. 5.54 ± 0.97, < 0.01), and Hes5 (2.89 ± 0.93 vs. 4.51 ± 1.54, < 0.01) in CD14(+) monocytes of chronic hepatitis C patients. Interaction of Notch signaling pathway with TLR4 can promote the function of CD14(+) monocytes in chronic hepatitis C patients.
观察慢性丙型肝炎患者Notch信号通路及Toll样受体4（TLR4）的表达变化及其相互作用对CD14(+)单核细胞功能的影响。选取2017年8月至10月就诊于陕西省人民医院的24例未经治疗的慢性丙型肝炎患者及10例健康个体。选用CD14(+)单核细胞，用Notch信号通路抑制剂DAPT刺激或转染TLR4 siRNA，采用实时定量PCR检测Notch1、Notch2、Hes1和Hes5 mRNA水平。采用蛋白质印迹法检测TLR4蛋白水平及NF-κB磷酸化水平。采用酶联免疫吸附测定法检测CD14(+)单核细胞分泌的细胞因子水平。采用t检验或配对t检验进行组间比较。慢性丙型肝炎患者CD14+单核细胞中Notch1 mRNA的相对表达量（3.97±?2.03比0.91±0.76，P<0.01）及Notch信号通路下游分子（5.96±2.31比0.99±0.45，P<0.01）、Hes1 mRNA和Hes5 mRNA（4.31±1.05比0.84±0.20，P<0.01）显著高于健康个体。慢性丙型肝炎患者CD14(+)单核细胞中TLR4 mRNA的相对表达量（5.14±1.09比1.27±0.39）及蛋白水平显著高于健康个体（P<0.01）。用DAPT抑制Notch信号通路可降低慢性丙型肝炎患者CD14(+)单核细胞中TLR4 mRNA的相对表达量（2.58±1.3?6比4.34±1.88，P<0.05）、蛋白表达及NF-κB磷酸化水平。此外，MCP-1分泌水平[（94.32±23.59）pg/ml比（64.07±9.39）pg/ml，P<0.01]及IL-8[（12.54±4.89）pg/ml比（7.92±3.01）pg/ml，P<0.05]显著降低。转染TLR4 siRNA可降低慢性丙型肝炎患者CD14(+)单核细胞中Notch1 mRNA（2.09±1.72比3.73±1.75，P<0.05）、Hes1（2.87±0.84比5.54±0.97，P<0.01）及Hes5（2.89±0.93比4.51±1.54，P<0.01）的表达。Notch信号通路与TLR4相互作用可促进慢性丙型肝炎患者CD14(+)单核细胞的功能。
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