Effect Of Intestinal Fungal Dysbiosis On Corneal Wound Healing In Mice

Object:To investigate the effect of intestinal fungal dysbiosis induced by antifungal drug on corneal wound healing in mice.Methods:80 male C57BL/6J mice without eye disease were selected under slit lamp microscope and divided into two groups randomly: amphotericin B treated(Amph)group and control(Ctrl)group.The Amph group was supplemented with amphotericin B to induce intestinal fungal dysbiosis and the Ctrl group was given a normal diet.After 4 weeks of feeding,fecal samples of two groups of mice were collected for intestinal fungal ITS sequencing,and corneal epithelial trauma was performed on the two groups of mice after identifying the disorder of intestinal fungal flora.Mice were anesthetized,and the central corneal epithelium was demarcated with a 2-mm trephine and then removed the corneal epithelium mechanically using a Golf Club Spud under a dissecting microscope.After the trauma,corneal epithelium was stained with 2% sodium fluorescein solution to evaluate the corneal wound area dynamically.Immunofluorescence staining was used to observe and quantitatively count the changes of corneal epithelial dividing cells,??T cells and neutrophils.HE staining was used to assess the change of corneal thickness.Results:Compared with Ctrl group: 1.Amphotericin B treatment resulted in dysbiosis of intestinal fungal in mice.2.The number of dividing cells in the Amph group decreased at 0h,6h,12 h,18h,24 h,30h,36 h and 42 h after trauma compared with that in the control group and the rate of reepithelization and wound repair were delayed.P < 0.05 at each time point,the difference was statistically significant.3.??T cells and neutrophils were significantly decreased in the Amph group,P < 0.05 was compared between the two groups.4.At 24 h,48h and 96 h after corneal trauma,the thickness of corneal epithelium in the Amph group became thinner,all P < 0.05,and the differences at each time point were statistically significant.Conclusion:Intestinal fungal dysbiosis delays the rate of reepithelization after corneal wound healing of mice,leading to a weaker inflammatory response,reducing the ability of repair after corneal trauma,and thins the thickness of corneal epithelium.