Li Yongwei, Chen Lu, Wang Chunxia, Chen Jianshe, Zhang Xiaoqian, Hu Yue, Niu Xiaobin, Pei Dongxu, He Zhiqiang, Bi Yongyi
Wuhan University School of Public HealthWuhan 430071, China; Department of Clinical Laboratory, Henan Province Hospital of Traditional Chinese MedicineZhengzhou 450002, China.
Beijing University of Chinese Medicine Beijing 100029, China.
Am J Transl Res. 2016 Dec 15;8(12):5619-5627. eCollection 2016.
This study aims to explore the role of extra-cellular matrix metalloproteinase inducer (EMMPRIN) in the drug resistance of the pseudomonas aeruginosa (PA). The BALB/c mice were transfected with PA, then the mice were infected with the siRNA of EMMPRIN to silence the EMMPRIN gene. The EMMPRIN mRNA and protein were detected by using RT-PCR and western blot, respectively. In order to examine the function of EMMPRIN in drug resistance of PA, the BALB/c and C57BL/6 mice were treated with EMMPRIN siRNA. The cytokines, EMMPRIN and MMP9 were examined by the RP-PCR and ELISA, respectively, undergoing the silence of EMMPRIN siRNA. Moreover, the western blot assay was also used to test the phosphorylated MAPK in the murine macrophages after silenced by the EMMPRIN siRNA. The EMMPRIN was activated, with lipopolysaccharide stimulation and treated with the MAPK inhibitor, to evaluate whether the MAPK participates in the EMMPRIN-triggered drug resistance. The results indicated that the EMMPRIN expression was elevated in the infected BALB/c at 3 or 5 days post-infection. Silence of EMMPRIN Enhanced the Production of pro-inflammatory cytokines in PA keratitis. Silence of EMMPRIN significantly up-regulated Th1-type cytokines IFN-γ, IL-12, and IL-18, but down-regulated Th2-type cytokines IL-4, IL-5, and IL-10. MMP9 was increased in the cells with rEMMPRIN treatment. EMMPRIN inhibits pro-inflammatory cytokine production via a MAPK signaling pathway. In conclusion, EMMPRIN promotes host resistance against pseudomonas aeruginosa infection via MAPK signaling pathway.
本研究旨在探讨细胞外基质金属蛋白酶诱导剂（EMMPRIN）在铜绿假单胞菌（PA）耐药性中的作用。将PA转染到BALB/c小鼠体内，然后用EMMPRIN的小干扰RNA（siRNA）感染小鼠，使EMMPRIN基因沉默。分别采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测EMMPRIN的信使核糖核酸（mRNA）和蛋白质。为了研究EMMPRIN在PA耐药性中的功能，用EMMPRIN siRNA处理BALB/c和C57BL/6小鼠。分别采用实时荧光定量聚合酶链反应（RP-PCR）和酶联免疫吸附测定（ELISA）检测在EMMPRIN siRNA沉默后细胞因子、EMMPRIN和基质金属蛋白酶9（MMP9）。此外，还采用蛋白质免疫印迹法检测EMMPRIN siRNA沉默后小鼠巨噬细胞中磷酸化的丝裂原活化蛋白激酶（MAPK）。用脂多糖刺激激活EMMPRIN并用MAPK抑制剂处理，以评估MAPK是否参与EMMPRIN引发 的耐药性。结果表明，感染后3天或5天，感染的BALB/c小鼠中EMMPRIN表达升高。EMMPRIN沉默增强了PA角膜炎中促炎细胞因子的产生。EMMPRIN沉默显著上调1型辅助性T细胞（Th1）型细胞因子干扰素-γ（IFN-γ）、白细胞介素-12（IL-12）和白细胞介素-18（IL-18），但下调2型辅助性T细胞（Th2）型细胞因子白细胞介素-4（IL-4）、白细胞介素-5（IL-5）和白细胞介素-10（IL-10）。用重组EMMPRIN（rEMMPRIN）处理的细胞中MMP9增加。EMMPRIN通过MAPK信号通路抑制促炎细胞因子的产生。总之，EMMPRIN通过MAPK信号通路促进宿主抵抗铜绿假单胞菌感染。
