Antifungal and Anti-inflammatory Effect of Punicalagin on Murine Aspergillus fumigatus Keratitis

ABSTRACT

Purpose

This study aimed to investigate the anti-inflammatory effect and antifungal effect of punicalagin in murine fungal keratitis.

Methods

We used in vitro and in vivo protocols to assess the anti-inflammatory effect and antifungal effect of punicalagin. In vitro, time kill and mycelial stain were done. In vivo, murine fungal keratitis was established and treated with PBS or PUN. Clinical scores were taken on days 1, 3, and 5 post infection. The mRNA and protein levels of inflammatory factors were detected by RT-PCR and Western blot, and the number and location of macrophages were analyzed by flow cytometry and immunofluorescence. Also, fungal plate counting was used to assess the antifungal effect. The DCFH-DA fluorescence probe detected the ROS level.

Results

In vitro, PUN showed activity against A.fumigatus. (A.F.), with MIC90 values of 250 μg/ml, and significantly reduced A.F. biofilm formation (p < .001). In vivo, the mouse fungal keratitis model after punicalagin treatment exhibited less disease, lower clinical scores (p < .05), lower reduced macrophage infiltrate (p < .001), and fungal load (p < .001) than those treated with PBS. Treatment with punicalagin also reduced the mRNA expression and protein level of pro-inflammatory factors. At the cellular level, PUN significantly reduced the mRNA expression of inflammatory factors and ROS production caused by the stimulation of mycelia in RAW264.7 (p < .001).

Conclusions

The results show that punicalagin is beneficial in the treatment of murine fungal keratitis. The mechanism of its anti-inflammatory effect was synthetical, including antifungal activity, an inhibitory effect of proinflammatory factor and macrophages, and anti-oxidation.

KEYWORDS:

• Fungal keratitis
• Punicalagin
• A.fumigatus
• mice
• macrophage

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article or its supplementary materials.

Other data that support the findings of this study are available from the corresponding author, Guiqiu Zhao, upon reasonable request.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the National Natural Science Foundation of China [no. 89100824].