Gene transfer of kringle 5 of plasminogen by electroporation inhibits corneal neovascularization.

To test the efficacy of naked plasmid that expresses human kringle 5 of plasminogen (K5) in suppressing experimental corneal neovascularization in a rat model. A eukaryotic expression plasmid encoding human K5 (pSecK5) was constructed. COS cells were transiently transfected with pSecK5 using a lipid-based transfection reagent. K5 secretion was confirmed by Western blot analysis. The effect of the secreted K5 on the proliferation of human umbilical vein endothelial cells (HUVECs) was investigated colorimetrically. Forty-three Sprague-Dawley rats were used for a corneal neovascularization suppression experiment. Corneal injury was induced by placing a disk of filter paper (immersed in 1 mol/l NaOH, 3.0 mm in diameter) on the corneal surface for 2 min. The cornea was immediately washed with saline. pSecK5 and empty plasmids were injected subconjunctivally, and square-wave electric pulses were immediately applied to the eyes. The expression of K5 was analyzed by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The extent of corneal neovascularization was evaluated by scores. The constructed plasmid could express itself in COS cells. Conditioned medium from K5-transfected COS cells apparently inhibited HUVEC proliferation, compared with conditioned medium from COS cells transfected with empty plasmid or nontransfected cells. RT-PCR and immunohistochemistry confirmed the expression of K5 in the conjunctiva and cornea. Corneal neovascularization was significantly suppressed by K5 gene transfer in rats' eyes. In a rat model, K5 gene transfer by subconjunctival injection and electroporation can effectively inhibit corneal neovascularization induced by an alkali burn.

Yu WZ ，Li XX ，She HC ，He PY ，Dong JQ ，Rui M ，Ma DL ... -  《OPHTHALMIC RESEARCH》

Acidic/neutral amino acid residues substitution in NH2 terminal of plasminogen kringle 5 exerts enhanced effects on corneal neovascularization.

Li C ，Li L ，Cheng R ，Dai Z ，Li C ，Yao Y ，Zhou T ，Yang Z ，Gao G ，Yang X ... -  《-》

Plasminogen kringle 5 inhibits alkali-burn-induced corneal neovascularization.

Plasminogen kringle 5 (K5) is a potent angiogenic inhibitor. The purpose of the present study was to evaluate the therapeutic effect of K5 on alkali-burn-induced corneal neovascularization (NV) and to investigate its mechanism of action. Corneal NV was induced in rabbits by NaOH. The rabbits received eye drops containing K5 or vehicle alone, four times per day. Corneal NV and inflammation were monitored every other day with a slit lamp microscope, and the length of the vessels in the cornea and the area of NV were measured. Vascular endothelial growth factor (VEGF) was determined by immunohistochemical and Western blot analyses. The TUNEL assay was used to assess the apoptosis of endothelial cells. The effects of K5 on primary bovine aortic endothelial cells (BAECs) were determined by MTT assay, flow cytometry, transmission electron microscopy, and DNA fragmentation assay. Alkali-burn-induced progressive corneal NV and inflammation in the cornea. K5 delayed the onset of corneal NV (P < 0.05) and decreased NV areas (P < 0.05) in a dose-dependent manner. K5 treatment, after the formation of corneal NV, induced regression of newly formatted vessels in the cornea. K5 decreased the inflammatory index in the corneas at different time points after the alkali burn. Corneal VEGF levels were reduced by K5 treatment. K5 inhibits proliferation and induces apoptosis in BAECs. Topical application of K5 may have therapeutic potential for the chemical burn-induced corneal NV and inflammation. The inhibitory effect of K5 on corneal NV may be by downregulation of VEGF expression.

Zhang Z ，Ma JX ，Gao G ，Li C ，Luo L ，Zhang M ，Yang W ，Jiang A ，Kuang W ，Xu L ，Chen J ，Liu Z ... -  《INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE》

Inhibition of corneal neovascularization with plasmid pigment epithelium-derived factor (p-PEDF) delivered by synthetic amphiphile INTeraction-18 (SAINT-18) vector in an experimental model of rat corneal angiogenesis.

Kuo CN ，Yang LC ，Yang CT ，Lai CH ，Chen MF ，Chen CY ，Chen CH ，Wu PC ，Kou HK ，Chen YJ ，Hung CH ，Tsai CB ... -  《-》

Corneal transduction to inhibit angiogenesis and graft failure.

Murthy RC ，McFarland TJ ，Yoken J ，Chen S ，Barone C ，Burke D ，Zhang Y ，Appukuttan B ，Stout JT ... -  《INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE》