The anti-inflammatory regulation of calcitonin gene-related peptide in mouse Aspergillus fumigatus keratitis
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Cui Li and Gui-Qiu Zhao. Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong Province, China. yankelicui@126.com; zhaoguiqiu_good@126.com

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Supported by National Natural Science Foundation of China (No.81870632); the Youth National Natural Science Foundation of China (No.81800800; No.81700800); the Natural Science Foundation of Shandong Province (No.ZR2017BH025).

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    Abstract:

    AIM: To analyze the impact of calcitonin gene-related peptide (CGRP) in mouse keratitis after Aspergillus fumigatus (A. fumigatus) infection. METHODS: C57BL/6 mice were treated subconjunctivally with different concentrations of exogenous CGRP, and BALB/c mice were treated with CGRP8-37 (a CGRP antagonist) before corneas were infected with A. fumigatus. The cornea was assessed under the slit-lamp and the clinical score was recorded. The mRNA levels of IL-1β, TNF-α, IL-6, and MIP-2 were detected by quantitative real-time polymerase chain reaction (PCR), while the protein level of IL-1β was determined by Western blotting. In vitro, RAW264.7 cells were used to investigate NLRP3 and IL-1β expression induced by A. fumigatus after the pretreatment of exogenous CGRP or CGRP8-37. Cytokines expression in RAW264.7 cells was evaluated by real-time PCR and Western blotting. RESULTS: Using exogenous CGRP resulted in down-regulated synthesis of IL-1β and MIP-2 stimulated by A. fumigatus in C57BL/6 mice keratitis, and the synthesis of IL-1β, MIP-2 and IL-6 was up-regulated in BALB/c mice corneas after the pretreatment with CGRP8-37. Pretreatment with exogenous CGRP and CGRP8-37 did not influence TNF-α mRNA levels either in BALB/c or C57BL/6 mice keratitis. The levels of NLRP3 and IL-1β were both reduced in A. fumigatus stimulated-macrophages after treatment with exogenous CGRP. And CGRP8-37 pretreatment would increase NLRP3 and IL-1β levels. CONCLUSION: CGRP may alleviate the inflammatory reaction in mice keratitis after infection with A. fumigatus. The anti-inflammatory effect may be related to the inhibition of NLRP3 expression by CGRP.

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Ming Xu, Cui Li, Gui-Qiu Zhao, et al. The anti-inflammatory regulation of calcitonin gene-related peptide in mouse Aspergillus fumigatus keratitis. Int J Ophthalmol, 2020,13(5):701-707

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Publication History
  • Received:November 12,2019
  • Revised:March 09,2020
  • Adopted:
  • Online: March 30,2020
  • Published: