Basic And Clinical Studies Of Conjunctival Fibrosis After Trabeculectomy

Part 1 The effect of thrombospondin-1 on the prognosis of trabeculectomyPurpose:To evaluate whether the level of thrombospondin-1(TSP-1)in aqueous humor can predict the prognosis of trabeculectomy in patients with primary angle closure glaucoma(PACG).Methods:This case-control study:involved 26 patients with PACG who experienced a failed trabeculectomy(case group)and 78 age-and sex-matched patients with PAGG who underwent successful trabeculectomy(control group).Aqueous humor was collected at the time of trabeculectomy and tested for TSP-1 levels with an en2yme-linked immunosorbent assay method.Logistic regression modeling was used to assess the risk factors for failed trabeculectomy.Results:The mean TSP-1 aqueous concentrations were significantly higher in the case group of 26 patients(20.67 ± 9.79 ng/ml)than the control group of 78 patients(5.17 ± 2.29 ng/ml)(P<0.001).Logistic regression analysis revealed TSP-1 level as an independent risk factor for a failed trabeculectomy.Conclusion:The aqueous humor TSP-1 level is higher in PACG eyes with failed trabeculectomy than with successful trabeculectomy at one year.The aqueous humor TSP-1 level is an independent risk factor associated with failed trabeculectomy.Part 2 Inhibitory effect of relaxin 2 on the activation of human Tenon’s fibroblasts induced by TGF-β1 and its mechanismPurpose:The aim of this study was to investigate the effect of relaxin 2 on proliferation,migration,and the production of extracellular matrix(ECM)protein of humanTenon’s fibroblasts(HTFs),and to assess the possible mechanism of it.Methods:Primarycultured HTFs,obtained from patientsundergoing strabismus surgery,were treated with relaxin 2,TGF-β1,TGF-β1+relaxin 2,or vehicle control.Cell proliferation was measured using a Cell Counting Kit-8 assay.Cell migration was investigatedwith a scratch-wound assay.The expression ofa-SMA,MMP2,Collegen I,fibronectin and the phosphorylation of Smad wereestimated using reversetranscription-PCR,western blot,Enzyme-linked immunosorbent assay(ELISA)or immunofluorescenceanalysis.Results:Primary HTFswere culturedsuccessfully.Primary HTFs were characterized by immunocytofluorescence staining withvimentin,fibroblast surface protein,but not stain with cytokeratin.TGF-β1significantlyincreased the expression of a-SMA mRNA and protein inHTFs.TGF-β1 also significantly increased the synthesis of fibronectin and collagensin HTFs,increased the proliferation and migration of HTFs,and significantly decreased the synthesis of MMP2;however relaxin2 treatment distinctly attenuated the TGF-β1-induced cell proliferation and migrationability of HTFsand production of ECMproteins.Relaxin2 also inhibited TGF-β1 induced the expression of the phosphorylation of Smad.Conclusions:Our resultsshowed that relaxin 2 attenuated TGF-β1-induced cell proliferation,migration,and ECM synthesis in HTFs.This function is s mainly by downregulating the relaxin 2/TGF-β/Smad/MMP2/collagenand fibronectin signaling pathways.This finding implies that relaxin 2 may be helpful in reducing the bleb fibrosis.