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Updated: Oct 27, 2025
In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model
Published on: January 21, 2018
Inhibitory effects of safranal on laser-induced choroidal neovascularization and human choroidal microvascular
Qin-Xiao1, Yao-Yao Sun2, Zhan-Jun Lu1
1Department of Ophthalmology, Affiliated Hospital of Inner Mongolia University for Nationalities, Tongliao 028007, Inner Mongolia Autonomous Region, China.
Aim:
To determine the effects of safranal on choroidal neovascularization (CNV) and oxidative stress damage of human choroidal microvascular endothelial cells (HCVECs) and its possible mechanisms.
Methods:
Forty-five rats were used as a laser-induced CNV model for testing the efficacy and safety of safranal (0.5 mg/kg·d, intraperitoneally) on CNV. CNV leakage on fluorescein angiography (FA) and CNV thickness on histology was compared. HCVECs were used for a H2O2-induced oxidative stress model to test the effect of safranal in vitro. MTT essay was carried to test the inhibition rate of safranal on cell viability at different concentrations. Tube formation was used to test protective effect of safranal on angiogenesis at different concentrations. mRNA transcriptome sequencing was performed to find the possible signal pathway. The expressions of different molecules and their phosphorylation level were validated by Western blotting.
Results:
On FA, the average CNV leakage area was 0.73±0.49 and 0.31±0.11 mm2 (P=0.012) in the control and safranal-treated group respectively. The average CNV thickness was 127.4±18.75 and 100.6±17.34 µm (P=0.001) in control and safranal-treated group. Under the condition of oxidative stress, cell proliferation was inhibited by safranal and inhibition rates were 7.4%-35.4% at the different concentrations. For tube formation study, the number of new branches was 364 in control group and 35, 42, and 17 in 20, 40, and 80 µg/mL safranal groups respectively (P<0.01). From the KEGG pathway bubble graph, the PI3K-AKT signaling pathway showed a high gene ratio. The protein expression was elevated of insulin receptor substrate (IRS) and the phosphorylation level of PI3K, phosphoinositide-dependent protein kinase 1/2 (PDK1/2), AKT and Bcl-2 associated death promoter (BAD) was also elevated under oxidative stress condition but inhibited by safranal.
Conclusion:
Safranal can inhibit CNV both in vivo and in vitro, and the IRS-PI3K-PDK1/2-AKT-BAD signaling pathway is involved in the pathogenesis of CNV.
Insights
Safranal effectively inhibits choroidal neovascularization (CNV) in both animal models and cell cultures. The study reveals safranal
Area of Science:
- Ophthalmology
- Molecular Biology
- Pharmacology
Background:
- Choroidal neovascularization (CNV) is a major cause of vision loss.
- Oxidative stress plays a significant role in the pathogenesis of CNV.
- Understanding the molecular mechanisms underlying CNV is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the therapeutic effects of safranal on CNV.
- To evaluate safranal's impact on oxidative stress in human choroidal microvascular endothelial cells (HCVECs).
- To elucidate the underlying molecular signaling pathways involved in safranal's action.
Main Methods:
- A laser-induced CNV rat model was used to assess safranal's efficacy and safety.
- HCVECs were subjected to hydrogen peroxide (H2O2)-induced oxidative stress for in vitro studies.
- Cell viability (MTT assay), tube formation, and mRNA transcriptome sequencing were performed.
- Western blotting was employed to validate key protein expressions and phosphorylation levels.
Main Results:
- Safranal significantly reduced CNV leakage and thickness in vivo.
- In vitro, safranal inhibited cell proliferation and tube formation in a dose-dependent manner.
- Transcriptome analysis identified the PI3K-AKT signaling pathway as a key target.
- Safranal modulated the expression and phosphorylation of proteins within the IRS-PI3K-PDK1/2-AKT-BAD pathway.
Conclusions:
- Safranal demonstrates significant inhibitory effects on CNV progression, both in vivo and in vitro.
- The IRS-PI3K-PDK1/2-AKT-BAD signaling pathway is implicated in CNV pathogenesis and is a target of safranal.
- Safranal holds potential as a therapeutic agent for treating CNV.
