Tat PTD–endostatin: A novel anti-angiogenesis protein with ocular barrier permeability via eye-drops

Autor: Fengshan Wang, Yi Qu, Haining Tan, Zhiwei Li, Yan-Na Cheng, Guoying Mu, Yan Li, Xinke Zhang
Rok vydání: 2015
Předmět:
Male
Time Factors
genetic structures
Angiogenesis
Recombinant Fusion Proteins
Biophysics
Neovascularization
Physiologic
Administration
Ophthalmic
Angiogenesis Inhibitors
Chick Embryo
macromolecular substances
Biology
Eye
Biochemistry
Chorioallantoic Membrane
Permeability
Cell Line
Flow cytometry
medicine
Animals
Molecular Biology
Cell Proliferation
Retina
Dose-Response Relationship
Drug
medicine.diagnostic_test
Cell growth
Endothelial Cells
Biological Transport
Molecular biology
Choroidal Neovascularization
eye diseases
Endostatins
Cell biology
Mice
Inbred C57BL
Endothelial stem cell
Disease Models
Animal
Chorioallantoic membrane
Choroidal neovascularization
medicine.anatomical_structure
Intravitreal Injections
cardiovascular system
Pinocytosis
sense organs
Ophthalmic Solutions
medicine.symptom
Endostatin
Zdroj: Biochimica et Biophysica Acta (BBA) - General Subjects. 1850:1140-1149
ISSN: 0304-4165
Popis: Background Endostatin, a specific inhibitor of endothelial cell proliferation and angiogenesis, has been proved to have effects on ocular neovascular diseases by intraocular injection. In order to increase its permeability to ocular barriers and make it effective on fundus oculi angiogenesis diseases via non-invasive administration (eye drops), endostatin was fused to Tat PTD via a genetic engineering method. Methods Most of the Tat PTD– endostatin was expressed as inclusion bodies in Escherichia coli, so pure and active Tat PTD–endostatin was prepared by a series of operations, including inclusion body denaturation, refolding and chromatography. The anti-angiogenesis activity of Tat PTD–endostatin was investigated by cell proliferation experiments and chick embryo chorioallantoic membrane assay. In addition, its translocating ability and concrete entry mechanism into cells were also investigated by fluorescence microscope and flow cytometry. The penetrating ability to ocular barriers was also studied by immunohistochemistry. A mouse choroidal neovascularization model was established to investigate the pharmacodynamics of Tat PTD–endostatin. Results The obtained Tat PTD–endostatin had excellent anti-angiogenesis activity and was superior to Es in cellular translocating. Macropinocytosis may be the dominant route of entry of Tat PTD–endostatin into cells. Tat PTD–endostatin could cross ocular barriers and arrive at the retina after eye-drop administration. In addition, it displayed inhibitory effects on choroidal neovascularization via eye drops. Conclusions Tat PTD–endostatin possessed excellent ocular penetrating ability and anti-angiogenesis effects. General significance Tat PTD is a promising ocular delivery tool, and Tat PTD–endostatin is a potential drug for curing fundus oculi angiogenesis diseases.
Databáze: OpenAIRE
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