Experimental inhibition of corneal neovascularization by endostatin gene transfection in vivo
Ping Zhang1, Dezheng Wu, Jian Ge
1Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Chian. zping66@163.net.
Objective:
To investigate endostatin gene therapy of rat corneal neovascularization induced by acid cauterization.
Methods:
pBlast-hEndostatin and pBlast-Mcs were identified by digestion with Nhe Iand Sal I, by PCR reaction, by sequence, and then by alignment of PCR products with the gene Bank using NCBIBLAST software. They were then purified with QIAGEN Endofree plasmid maxi kit. Rat corneal neovascularization models were made with 75% AgNO(3) and 25% KNO(3) cauterization. The treatment method was subconjunctive injection of the pBlast-hEndostatin with the control of pBlast-Mcs.
Results:
pBlast-hEndostatin was found to contain the human endostatin gene. The rat corneal neovascularization induced by acid cauterization was significantly suppressed after subconjunctive injection of the pBlast-hEndostatin with inhibition rates of 37%, 40.2%, and 42.8% respectively on the sixth, tenth, and fifteenth day. The inhibition rate for the density of corneal neovascularization was 40%. However, no inhibition effect on the length of the neovascularization and corneal inflammatory cells was observed. Corneal neovascularization areas were positively correlated with edema and corneal opacity.
Conclusions:
The plasmid of pBlast-hEndostatin contained the human endostatin gene. The rat corneal neovascularization induced by acid cauterization can be partially inhibited by subconjunctive injection of the pBlast-hEndostatin mediated by liposomes. Endostatin produced by transfected fibroblast cells directly inhibits corneal neovascularization. This is not caused by inflammatory reaction inhibition.
Insights
Gene therapy using endostatin (pBlast-hEndostatin) partially inhibited rat corneal neovascularization. This novel approach shows promise for treating eye conditions by directly targeting abnormal blood vessel growth in the cornea.
Area of Science:
- Ophthalmology
- Gene Therapy
- Molecular Biology
Background:
- Corneal neovascularization is a significant cause of vision impairment.
- Acid cauterization is a common method for inducing experimental corneal neovascularization in rats.
- Gene therapy offers a potential therapeutic strategy for neovascular eye diseases.
Purpose of the Study:
- To evaluate the efficacy of endostatin gene therapy in a rat model of corneal neovascularization.
- To determine the inhibitory effect of pBlast-hEndostatin on acid-induced corneal neovascularization.
Main Methods:
- Construction and purification of pBlast-hEndostatin plasmid containing the human endostatin gene.
- Induction of corneal neovascularization in rats using a standardized acid cauterization method.
- Subconjunctival injection of pBlast-hEndostatin or control plasmid (pBlast-Mcs) for treatment.
Main Results:
- Successful identification and purification of the pBlast-hEndostatin plasmid.
- Significant suppression of corneal neovascularization density by 40% after endostatin gene therapy.
- Inhibition rates of 37-42.8% observed on days 6, 10, and 15.
- No significant effect on neovascularization length or corneal inflammatory cells.
Conclusions:
- The pBlast-hEndostatin plasmid effectively delivered the human endostatin gene.
- Subconjunctival injection of pBlast-hEndostatin partially inhibits experimental corneal neovascularization.
- Endostatin directly inhibits corneal neovascularization, independent of anti-inflammatory effects.
