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Peer-reviewedCC BY 4.0International Journal of Molecular Sciences

Recombinant Human Lysozyme Attenuates Hyperosmolarity-Induced Dry Eye Disease by Suppressing Oxidative Stress and Modulating NLRP3 Inflammasome Signaling

  • Yuqing Jiang1
  • Weili Hu1
  • Lingxiao Yi1
  • Minal Thacker1
  • Qiyun Wu2,3
  • Yuan Li2
  • En Gao2
  • Jingfang Bian1,3
  • Chi Wai Do1,3
  • Thomas Chuen Lam1,3
  • Man Sau Wong1,4,5
  • Liping Zhou1,2,5
5 affiliations
  1. 1Centre for Eye and Vision Research (CEVR), 17W Hong Kong, Science Park, Hong Kong
  2. 2PolyU–HuiKang Joint Laboratory, The Hong Kong Polytechnic University, Kowloon, Hong Kong
  3. 3School of Optometry, The Hong Kong Polytechnic University, Kowloon, Hong Kong
  4. 4Department of Food Science and Nutrition, The Hong Kong Polytechnic University, Kowloon, Hong Kong
  5. 5Research Centre for Chinese Medicine Innovation, The Hong Kong Polytechnic University, Kowloon, Hong Kong
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Abstract

Given the escalating global prevalence of dry eye disease (DED) and the inadequacy of current palliative management, this study was to evaluate thetherapeutic potential of recombinant lysozyme (rlysozyme) and investigate underlying mechanisms. Human corneal epithelial cells (HCEC) were subjected to hyperosmotic stress and treated with graded rlysozyme. Cell viability, wound healing, barrier integrity, oxidative stress, cytokines, and mitochondrial function were evaluated by MTS and scratch assays, immunofluorescent staining, ELISA, qRT-PCR, and flow cytometry. Results: No cytotoxicity was observed at any of the tested concentrations (2.76 × 10−9–2.76 × 10−4 M) in HCEC. In hyperosmolarity-induced DED cells, rlysozyme at the highest concentrations (2.76 × 10−4 M) significantly increased cell viability by 12.30% (p < 0.001) and promoted corneal repair by 45.04% (p < 0.05), which were comparable to those of 0.4% hyaluronic acid (the positive control). It also improved via restoring ZO-1 organization. Mechanistically, qRT-PCR results revealed that rlysozyme reduced mRNA expression of IL-1β (by 28.7%), TNF (by 39.2%) and key NLRP3 inflammasome components, including NLRP3 (by 62.0%) and PYD and CARD domain-containing protein gene(PYCARD, 21.4%), while caspase-1 (CASP1) and IL-6 expression remained unchanged. Moreover, oxidative stress was attenuated, as evidenced by significant decreases in reactive oxygen species (ROS, by 59.1%), 8-hydroxy-2-deoxyguanosine levels (8-OHdG, by 23.6%) and total matrix metalloproteinase-9 (MMP-9, by 64.3%) protein levels (p < 0.001), while mitochondrial membrane potential was assessed qualitatively. In conclusion, this study demonstrates thatrlysozyme effectively enhanced corneal epithelial cell viability, promoted wound healing, and restored ZO-1 structural organization without inducing cytotoxicity. These protective effects were associated with downregulation of NLRP3 inflammasome-related genes and attenuation of oxidative stress, supporting further investigation of its therapeutic potential in DED.

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Cite

BibTeXRIS
BibTeX
@article{jiang2026recombinant,
  title = {{Recombinant Human Lysozyme Attenuates Hyperosmolarity-Induced Dry Eye Disease by Suppressing Oxidative Stress and Modulating NLRP3 Inflammasome Signaling}},
  author = {Yuqing Jiang and Weili Hu and Lingxiao Yi and Minal Thacker and Qiyun Wu and Yuan Li and En Gao and Jingfang Bian and Chi Wai Do and Thomas Chuen Lam and Man Sau Wong and Liping Zhou},
  journal = {International Journal of Molecular Sciences},
  year = {2026},
  month = oct,
  doi = {10.3390/ijms27209297},
  url = {https://www.mdpi.com/1422-0067/27/20/9297},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Recombinant Human Lysozyme Attenuates Hyperosmolarity-Induced Dry Eye Disease by Suppressing Oxidative Stress and Modulating NLRP3 Inflammasome Signaling
AU  - Yuqing Jiang
AU  - Weili Hu
AU  - Lingxiao Yi
AU  - Minal Thacker
AU  - Qiyun Wu
AU  - Yuan Li
AU  - En Gao
AU  - Jingfang Bian
AU  - Chi Wai Do
AU  - Thomas Chuen Lam
AU  - Man Sau Wong
AU  - Liping Zhou
PY  - 2026
DA  - 2026/10/01
JO  - International Journal of Molecular Sciences
DO  - 10.3390/ijms27209297
UR  - https://www.mdpi.com/1422-0067/27/20/9297
AB  - Given the escalating global prevalence of dry eye disease (DED) and the inadequacy of current palliative management, this study was to evaluate thetherapeutic potential of recombinant lysozyme (rlysozyme) and investigate underlying mechanisms. Human corneal epithelial cells (HCEC) were subjected to hyperosmotic stress and treated with graded rlysozyme. Cell viability, wound healing, barrier integrity, oxidative stress, cytokines, and mitochondrial function were evaluated by MTS and scratch assays, immunofluorescent staining, ELISA, qRT-PCR, and flow cytometry. Results: No cytotoxicity was observed at any of the tested concentrations (2.76 × 10−9–2.76 × 10−4 M) in HCEC. In hyperosmolarity-induced DED cells, rlysozyme at the highest concentrations (2.76 × 10−4 M) significantly increased cell viability by 12.30% (p < 0.001) and promoted corneal repair by 45.04% (p < 0.05), which were comparable to those of 0.4% hyaluronic acid (the positive control). It also improved via restoring ZO-1 organization. Mechanistically, qRT-PCR results revealed that rlysozyme reduced mRNA expression of IL-1β (by 28.7%), TNF (by 39.2%) and key NLRP3 inflammasome components, including NLRP3 (by 62.0%) and PYD and CARD domain-containing protein gene(PYCARD, 21.4%), while caspase-1 (CASP1) and IL-6 expression remained unchanged. Moreover, oxidative stress was attenuated, as evidenced by significant decreases in reactive oxygen species (ROS, by 59.1%), 8-hydroxy-2-deoxyguanosine levels (8-OHdG, by 23.6%) and total matrix metalloproteinase-9 (MMP-9, by 64.3%) protein levels (p < 0.001), while mitochondrial membrane potential was assessed qualitatively. In conclusion, this study demonstrates thatrlysozyme effectively enhanced corneal epithelial cell viability, promoted wound healing, and restored ZO-1 structural organization without inducing cytotoxicity. These protective effects were associated with downregulation of NLRP3 inflammasome-related genes and attenuation of oxidative stress, supporting further investigation of its therapeutic potential in DED.
SN  - 1422-0067
ER  -