Xian Wu1, Guiqiu Zhao2, Jing Lin1, Nan Jiang1, Cui Li1, Liting Hu1, Xudong Peng1, Qiang Xu1, Qian Wang1, Hui Li1, Yan Zhang1. 1. Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China. 2. Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, China. Electronic address: guiqiuzhao@126.com.
Abstract
OBJECTIVE: To observe the production mechanism of surfactant protein D (SP-D) in human corneal epithelial cells (HCECs) when infected by Aspergillus fumigatus (A. fumigatus) hyphae, and explore whether SP-D can inhibit the cell activations through toll-like receptor 4 signaling pathway during fungal infection. METHODS: mRNA and protein expressions of SP-D were evaluated in HCECs after stimulation by A. fumigatus, with or without pretreatment of TLR4 inhibitor (CLI-095) by real time PCR and Western blot. The expression levels of inflammatory cytokines IL-1β and IL-8 evaluated when pretreated with SP-D antibody or recombinant human SP-D in fungi-stimulated HCECs by real time PCR and ELISA, IL-1β and IL-8 expressions were also detected in A. fumigatus-stimulated HCECs that pretreated with CLI095 or MyD88 inhibitor (Pepinh-MYD) and recombinant human SP-D. RESULTS: mRNA and protein levels of SP-D increased after stimulation of A. fumigatus for 16h and 20h respectively. The upregulation of SP-D could be inhibited by CLI-095. mRNA and protein expressions of IL-1β and IL-8 decreased significantly when pretreated HCECs with recombinant human SP-D for 4h before A. fumigatus stimulation, while IL-1β and IL-8 increased when pretreated with SP-D antibody for 1h. Pretreatment of CLI095 or Pepinh-MYD can increase the expressions of IL-1β and IL-8 mRNA and protein in HCECs induced by recombinant human SP-D and A. fumigatus. CONCLUSIONS: SP-D can be stimulated by TLR4 during A. fumigatus infection. Recombinant human SP-D can inhibit the expression of inflammatory cytokines through TLR4 signaling pathway.
OBJECTIVE: To observe the production mechanism of surfactant protein D (SP-D) in human corneal epithelial cells (HCECs) when infected by Aspergillus fumigatus (A. fumigatus) hyphae, and explore whether SP-D can inhibit the cell activations through toll-like receptor 4 signaling pathway during fungal infection. METHODS: mRNA and protein expressions of SP-D were evaluated in HCECs after stimulation by A. fumigatus, with or without pretreatment of TLR4 inhibitor (CLI-095) by real time PCR and Western blot. The expression levels of inflammatory cytokines IL-1β and IL-8 evaluated when pretreated with SP-D antibody or recombinant humanSP-D in fungi-stimulated HCECs by real time PCR and ELISA, IL-1β and IL-8 expressions were also detected in A. fumigatus-stimulated HCECs that pretreated with CLI095 or MyD88 inhibitor (Pepinh-MYD) and recombinant humanSP-D. RESULTS: mRNA and protein levels of SP-D increased after stimulation of A. fumigatus for 16h and 20h respectively. The upregulation of SP-D could be inhibited by CLI-095. mRNA and protein expressions of IL-1β and IL-8 decreased significantly when pretreated HCECs with recombinant humanSP-D for 4h before A. fumigatus stimulation, while IL-1β and IL-8 increased when pretreated with SP-D antibody for 1h. Pretreatment of CLI095 or Pepinh-MYD can increase the expressions of IL-1β and IL-8 mRNA and protein in HCECs induced by recombinant humanSP-D and A. fumigatus. CONCLUSIONS:SP-D can be stimulated by TLR4 during A. fumigatusinfection. Recombinant humanSP-D can inhibit the expression of inflammatory cytokines through TLR4 signaling pathway.
Authors: Yan Zhou; Jianghua Ming; Yaming Li; Xianjin Du; Ming Deng; Bin He; Jianlin Zhou; Guirong Wang; Shiqing Liu Journal: Biochem Biophys Res Commun Date: 2017-10-31 Impact factor: 3.575