Objective To investigate whether rAAV-NTF2 can decrease the permeability of outer blood-retina barrier and its possible effect on VEGF in high glucose condition in vitro.

Method RPE cell cultured from adult healthy donor eyes according to established method. NTF2 gene was cloned into adeno-associated virus vector pSNAV and we obtained high-titer rAAV2-NTF2, then, we estimated the transfection-positive RPE cell rate by laser confocal scanning microscope. The Outer blood-retina barrier model was constructed by human-RPE culture on Transwell membrane.  Tight junction (TJ) was observed with transmission electron microscopy (TEM) and evaluated by Transepithlial electrical resistance (TER) measurement and permeation rate of FITC-albumin .TJ associated proteins (Claudin1, Oclaudin, ZO-1) and VEGF were detected by Real-time PCR and Western blot and immunochemistry staining.
Result human-RPE cultured on Transwell membrane can form outer blood-retina barrier model successfully. After transfected rAAV-NTF2, the cells down-regulated VEGF expression and more TJ formed observed in TEM.  rAAV-NTF2 increased TER and decreased the permeability of outer blood-retina barrier significantly .TJ associated proteins (Claudin1, Oclaudin, ZO-1,etc) also have significantly different from the control.
conclusion  NTF2 can protect the outer blood-retina barrier from disfunction induced by high glucose in vitro.