Effects of arginine-glycine-aspartic acid peptide hydrogels on the activation of Tenon capsule fibroblasts and Yes-associated protein signaling
10.13389/j.cnki.rao.2024.0131
- VernacularTitle:RGD多肽水凝胶对Tenon囊成纤维细胞活化和YAP信号的影响
- Author:
Yao LÜ
1
;
Liang LIANG
;
Mian QIN
;
Ruping JIANG
;
Xia WU
Author Information
1. 443003 湖北省宜昌市,宜昌市中心人民医院眼科,三峡大学第一临床医学院
- Keywords:
Tenon capsule fibroblasts;
arginine-glycine-aspartic acid peptide hydrogel;
extracellular matrix;
Yes-as-sociated protein;
scar formation
- From:
Recent Advances in Ophthalmology
2024;44(9):686-691
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of arginine-glycine-aspartic acid(RGD)peptide hydrogels at differ-ent concentrations on the activation of Tenon capsule fibroblasts(HTF)and the expression of Yes-associated protein(YAP).Methods Three concentrations(0.5％,1.0％,2.0％)of RGD peptide hydrogels were prepared.Their internal microstructures were observed under a transmission electron microscope,and their elastic moduli(E)were measured using the rheological test.SD male rat models with conjunctival injury were established and divided into a blank group(no treatment),a low-concentration RGD group(subconjunctival injection of 0.5％RGD peptide hydrogel),a medium-con-centration RGD group(subconjunctival injection of 1.0％RGD peptide hydrogel),and a high-concentration RGD group(subconjunctival injection of 2.0％RGD peptide hydrogel).After one week,tissues were collected,and collagen fiber pro-liferation in the rat conjunctiva was observed after Masson staining.Immunohistochemistry was performed to detect the ex-pression of YAP and α-smooth muscle actin(α-SMA)in the rat conjunctiva.Scar formation cell models were constructed by stimulating HTF activation with transforming growth factor-β2(TGF-β2).According to experimental requirements,the cells were divided into a control group,a 0.5％RGD hydrogel group,a 1.0％RGD hydrogel group,a 2.0％RGD hydrogel group,and a normal group,and co-cultured for 24 hours.The relative expression levels of YAP,connective tissue growth factor(CTGF),α-SMA,fibronectin(FN),and type Ⅰ collagen(Col Ⅰ)proteins in HTF were detected by Western blot.The localization of cytoskeletal protein(F-actin)and YAP protein expression was observed under a confocal laser scanning mi-croscope.Results Electron microscopy showed that the crosslinking density of the nanofibers inside the RGD peptide hydrogel increased with the rise of the peptide concentration.Rheological measurements indicated that the E values of the 0.5％,1.0％,and 2.0％RGD peptide hydrogels were approximately 0.067 kPa,0.150 kPa,and 2.170 kPa,respectively.Masson staining results revealed that the area of collagen fiber proliferation in the conjunctiva of rats in the medium-concen-tration RGD group was significantly lower than that in the blank group(P＜0.05),while it showed no significant difference in the low-concentration and high-concentration RGD groups compared with the blank group(both P＞0.05).Immunohis-tochemistry results showed that the relative expression levels of YAP and α-SMA proteins in the conjunctiva of rats in the low-,medium-,and high-concentration RGD groups decreased compared to the blank group,and the decrease was the most significant in the medium-concentration RGD group(all P＜0.05).Cell immunofluorescence staining results indicated that the green fluorescence of F-actin was attenuated,and the red fluorescence of YAP was translocated from the nucleus to the cytoplasm in the 0.5％,1.0％,and 2.0％RGD hydrogel groups compared to the control group.Western blot results showed that the relative expression levels of YAP,CTGF,α-SMA,FN,and Col Ⅰ proteins in HTF were lower in the 0.5％,1.0％,and 2.0％RGD hydrogel groups compared to the control group,with the 1.0％RGD hydrogel group showing the most significant reduction(all P＜0.05).Conclusion With the optimal concentration of 1.0％,the RGD peptide hydro-gel can inhibit YAP protein expression and nuclear translocation and reduce HTF activation and the expression of fibrotic proteins,thus alleviating scar formation.
