Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: Corneal damage in T2DM mice is associated with deficient GDNF expression. Body weight ( A ) and blood glucose levels ( B ) were significantly elevated in T2DM mice ( n = 6). ( C ) Corneal mechanical sensitivity was markedly reduced in the diabetic group ( n = 6). Representative images ( D ) and quantitative clinical scores ( E ) demonstrating significantly increased corneal fluorescein staining in T2DM mice ( n = 12). ( F, G ) Expression levels of GDNF were significantly downregulated in the corneas of T2DM mice ( n = 6). ( H ) Quantitative analysis revealing a significant decrease in corneal epithelial thickness in T2DM mice ( n = 6). ** P < 0.01; *** P < 0.001; T2DM, type 2 diabetes mellitus.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Expressing, Sensitivity, Staining, Analysis

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF promotes proliferation and inhibits apoptosis in HCECs by modulating the PI3K/AKT pathway. ( A ) The proliferative viability of HCECs under diverse culture conditions, assessed via the CCK-8 assay ( n = 5). ( B, C ) Representative flow cytometry plots B and quantitative analysis C of HCEC apoptosis levels across different treatments ( n = 3). Representative micrographs ( D ) and quantification of the relative migratory capacity ( E ) of HCECs subjected to various culture conditions, as determined by the scratch wound assay ( n = 5). ns, not significant; * P < 0.05; *** P < 0.001; HG, high glucose; PA, palmitic acid.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: CCK-8 Assay, Flow Cytometry, Analysis, Scratch Wound Assay Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF alleviates ER stress in HCECs via activation of the PI3K/AKT signaling pathway. Representative Western blot bands ( A ) and corresponding quantitative analysis ( B ) of PI3K/AKT signaling pathway proteins and ER stress markers in HCECs across different culture conditions ( n = 3). ( C ) Representative transmission electron microscopy images revealing the ultrastructural morphological alterations of the ER in HCECs under the corresponding treatments. Magnified images of the boxed regions are shown in the right panels. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Activation Assay, Western Blot, Analysis, Transmission Assay, Electron Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF ameliorates corneal epithelial damage and accelerates wound healing in T2DM mice. ( A ) Representative ocular surface images showing corneal fluorescein staining and epithelial wound closure dynamics in mice under different in vivo interventions. ( B ) Quantitative statistical analysis of the corneal fluorescein staining scores ( n = 12). ( C ) Quantitative analysis of the relative corneal epithelial wound closure rates over time ( n = 5). *** P < 0.001; T2DM, type 2 diabetes mellitus.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Staining, In Vivo, Analysis

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF promotes corneal nerve regeneration and restores sensory function in T2DM mice. ( A ) Representative whole-mount immunofluorescence images of the corneal subbasal nerve plexus in mice across varying conditions. Scale bar = 500 µm. ( B ) Quantitative statistical analysis of the corneal nerve fiber density ( n = 6). ( C ) Evaluation and statistical analysis of corneal mechanical sensitivity among the experimental in vivo groups ( n = 6). ** P < 0.01; *** P < 0.001; T2DM, type 2 diabetes mellitus.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Immunofluorescence, Analysis, Sensitivity, In Vivo

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF attenuates corneal epithelial ER stress in T2DM mice via the PI3K/AKT pathway. ( A , B ) Protein expression levels of the PI3K/AKT signaling pathway components and ER stress markers in the murine corneas under varying in vivo conditions ( n = 3). ( C ) Representative transmission electron microscopy images illustrating the ultrastructural alterations of the ER within the murine corneal epithelium. Magnified images of the boxed regions are shown in the right panels. * P < 0.05; ** P < 0.01; T2DM, type 2 diabetes mellitus.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Expressing, In Vivo, Transmission Assay, Electron Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: GDNF mitigates corneal neural ER stress in T2DM mice through the PI3K/AKT pathway. Representative Western blot bands ( A ) and corresponding quantitative statistical analysis ( B ) of PI3K/AKT pathway proteins and ER stress markers in the trigeminal ganglia of mice subjected to different treatments ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001; T2DM, type 2 diabetes mellitus.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Western Blot, Analysis

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting Endoplasmic Reticulum Stress With Exogenous GDNF Promotes Diabetic Corneal Epithelial and Neural Repair via PI3K/AKT Signaling

doi: 10.1167/iovs.67.12.24

Figure Lengend Snippet: Mechanistic paradigm of GDNF mediated ocular surface repair. GDNF rescues delayed epithelial surface regeneration and stimulates nerve regrowth within the diabetic cornea. These profound therapeutic outcomes are mechanistically driven by the suppression of endoplasmic reticulum stress via PI3K/AKT pathway engagement. ER, endoplasmic reticulum.

Article Snippet: To mimic the T2DM microenvironment in vitro, HCECs were exposed to glucolipotoxic conditions for 48 hours using a medium supplemented with 30 mM glucose and 300 μM palmitic acid (PA; Proteintech, China), either in the presence or absence of exogenous GDNF (100 ng/mL; MedChemExpress).

Techniques: Sensor Regeneration