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Tear film instability is associated with weakened colocalization between occludin and MUC5AC in scopolamine-induced dry eye disease (DED) rats

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Abstract

Purpose

Dry eye disease (DED) is a disease with tear film instability because of multiple factors. This study was conducted to explore roles of occludin and MUC5AC in tear film instability in DED rat model.

Methods

A total of 20 SD rats were divided into DED group (n = 10) and normal control (NC) group (n = 10). DED rat model was established by subcutaneously injecting with scopolamine hydrobromide. Clinical examinations, including tear breakup time (tBUT), Schirmer’s test and corneal fluorescein staining, were conducted to determine corneal functions. Transmission electron microscopy was used to measure the ultrastructures of corneal epithelial cells. Western blotting assay was used to identify occludin expression in corneal tissues of DED rats. Real-time PCR (RT-PCR) was performed to verify gene transcription of occludin and MUC5AC. Colocalization between occludin and MUC5AC was identified with confocal fluorescence microscopy.

Results

Tear breakup time was significantly shorter, and corneal fluorescein staining score was predominantly higher in DED rats compared to those in normal rats (P < 0.05). Normal rats showed a steady tear secretion throughout the whole experiments, while DED rats showed a dramatic reduction on day 14. DED rats demonstrated ultrastructural damage of Golgi apparatus and endoplasmic reticulum in corneal epithelial cells. Occludin and MUC5AC expressions were significantly downregulated in corneal tissue of DED rats compared with those of normal rats (P < 0.05). Percentage of occludin-MUC5AC-colocalized corneal epithelial cells in DED rats was significantly less compared with those in normal rats (P < 0.01).

Conclusions

Tear film stability was damaged in scopolamine-induced DED rats because of the weakened colocalization between occludin and MUC5AC molecule. This study would provide a potential clue for the pathogenesis and a promising theoretical basis for clinical work of DED.

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Funding

This study was granted by the National Natural Science Foundation of China (Youth Program) (Grant No. NSFC81400382), Shaanxi Innovation Capability Support Program (Grant No. 2016KJXX-82) and Scientific Research Talent Project of Xi'an Health Commission (Grant No. 2022yb19).

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Authors and Affiliations

  1. Xi’an Center for Disease Control and Prevention, No. 599, Xiying Road, Xi’an, 710054, China

    Kun Xu

  2. Xi’an No. 1 Hospital, Shaanxi Institute of Ophthalmology, Shaanxi Key Laboratory of Ophthalmology, Clinical Research Center for Ophthalmology Diseases of Shaanxi Province, The First Affiliated Hospital of Northwest University, Xi’an, China

    Kun Xu, Xian-ning Liu, Hong-bing Zhang, Xiu-ping Zhu & Xian-jiao Zhang

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  1. Kun Xu
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Contributions

All authors read and approved final manuscript. XK had the idea for article, drafted the manuscript, performed literature, critically revised manuscript and made the final approval; XNL, HBZ, XPZ and XJZ performed the literature search and data analysis.

Corresponding author

Correspondence to Kun Xu.

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The authors declare no competing interests.

Ethical approval

This research has been approved by the Ethical Committee of the Xi’an Center for Disease Control and Prevention (Approval No. 2021-001). This study was performed based on the guidance of Declaration of Helsinki.

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Xu, K., Liu, Xn., Zhang, Hb. et al. Tear film instability is associated with weakened colocalization between occludin and MUC5AC in scopolamine-induced dry eye disease (DED) rats. Int Ophthalmol 43, 463–473 (2023). https://doi.org/10.1007/s10792-022-02443-x

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