Rosiglitazone prevents fibrosis after glaucoma filtration surgery by promoting autophagy: in vitro and in vivo

Abstract

Purpose : To investigate the influence of rosiglitazone on fibrosis after glaucoma filtration surgery (GFS) through the experiment of human Tenon’s fibroblasts (HTFs) and rabbit GFS model to access the possible mechanism.

Methods : Cultured human Tenon’s fibroblasts were pretreated in rosiglitazone before being stimulated with transforming growth factor β1 (TGF-β1). Expression of associated targets of autophagy and fibrosis were examined by real-time quantitative RT-PCR, western blot analysis (WB), transmission electron microscope (TEM), immunofluorescence. Trabeculectomy has performed on 30 rabbits that intraocular pressure was in a stable range of 25mmHg to 35mmHg. The operated eyes were followed up within 28 days. After the surgery, immunohistochemistry was performed on D14 and D28, Western blot analysis was performed on D28.

Results : The experimental group had the least expression of Collagen1, SP1, CTGF, α-SMA, P62 and higher expression of Beclin1, LC3, Vps34, and Atg14L. After GFS on rabbits eyes, the IOP of was significantly lower. The level of Collagen1, SP1, CTGF, α-SMA expression also was reduced in the sclera and conjunctival areas in the rosiglitazone group.

Conclusions : Rosiglitazone can inhibit the formation of scarring though activate autophagy by Beclin1/Vps34 pathway

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

HTFs were transfected by SensGFP-StubRFP-LC3 lentivirus (green and red) and were stained by Hochest (blue). The green pots stand for the number of autophagosomes and the red pots stand for the number of autolysosomes. The TGF-β1 group has more red pots than the control group. Also, rosiglitazone + TGF-β1 and rapamycin + TGF-β1 group significantly have much more green and red pots than the vehicle group (DMSO+ TGF-β1 group).

 

Appearances of filtration blebs in the four groups 7 days, 14 days, 21 days and 28 days after GFS. 7 days after surgery, a diffuse bleb was observed in each eye of four groups. In the surgery only group, bleb was gradually disappeared. It is very apparent that the bleb was totally scarring in the surgery the only group on 21 days after GFS. This phenomenon can also find in the group of 5-fu. However, in the groups of RGS and MMC, the blebs were also exited on 28 days after GFS.