ObjectiveTo observe the expression of heat shock protein 47 (HSP47) and transforming growth factor-β2(TGF-β2) in vitreous specimens and epiretinal membranes of patients with proliferative vitreoretinopathy diseases. MethodsVitreous specimens and epiretinal membranes were obtained from 48 patients (48 eyes) with proliferative vitreoretinopathy (PVR) and 50 patients (50 eyes) with proliferative diabetic retinopathy (PDR). Vitreous specimens and internal limiting membranes were collected from 20 patients (20 eyes) with idiopathic macular hole (IMH) as control group. The expression of HSP47 and TGF-β2 in the vitreous specimens was evaluated using enzyme linked immunosorbent assay. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in epiretinal membrane and internal limiting membrane specimens were observed for immunohistochemical staining method. The correlation between the positive expression of HSP47 and TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR and PDR were analyzed. ResultsThe expression of HSP47 in vitreous specimens of patients with PVR, PDR and IMH were (212.35±23.32), (231.30±26.79), (171.06±28.91) pg/ml, respectively. The expression of TGF-β2 in vitreous specimens of patients with PVR, PDR and IMH were (1919.96±318.55), (1939.39±177.57), (1194.61±234.20) pg/ml, respectively. The expression of HSP47, TGF-β2 in the vitreous specimens of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=12.952, 34.532;P < 0.01). The epiretinal membrane of patients with PVR and PDR showed markedly increased expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the cytoplasm and extracellular matrix. The expression of HSP47 and typeⅢcollagen was negative and the expression of TGF-β2 was weakly positive and the expression of typesⅠcollagen was positive in internal limiting membrane of patients with IMH. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the epiretinal membrane of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=13.469, 18.752, 12.875, 20.358; P < 0.01). The expression of HSP47 was positively correlated with the positive expression of TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR (r=0.475, 0.556, 0.468; P < 0.05) and PDR (r=0.484, 0.589, 0.512; P < 0.05). ConclusionsThis study showed increased consistent expression of HSP47 and TGF-β2 in vitreous and epiretinal membrane specimens of patients with PVR and PDR. Both HSP47 and TGF-β2 were expressed in the cytoplasm and extracellular matrix. HSP47 and TGF-β2 may be involved in the pathological process of PDR and PVR by promoting collagen synthesis.
ObjectiveTo observe the expression of hot shock protein 47 (HSP47) in pre-retinal membrane of proliferative vitreoretinopathy (PVR) and the influence of transforming growth factor-β2 (TGF-β2) on the expression of HSP47 in retinal pigment epithelial (RPE) cell. MethodsPre-retinal membranes were collected and observed by hematoxylin-eosin, Masson and immunohistochemical staining. Cultured ARPE-19 cells were treated with TGF-β2 at serial concentration (0, 1, 5, 10 ng/ml) and time (0, 12, 24, 48 hours), respectively. And then the mRNA and protein expressions of HSP47 and Col-Ⅰ were measured by fluorescence quantitative reverse transcription polymerase chain reaction and Western blot at the same time. ResultsA lot of epithelial cells with pigmental particles were observed in pre-retinal membranes of PVR, much accumulated collagen protein was observed in the specimens, and HSP47 positive expression was bserved in cytoplasm and stroma of most of the epithelioid cells. Compared with 0 ng/ml group, the expressions of HSP47 mRNA in ARPE-19 were up-regulated by 1.32, 2.35, 1.85 fold, significant differences were observed in all groups (F=27.21, P<0.05); the expressions of protein were up-regulated by 2.33, 2.89, 2.60 fold, significant differences were observed in all groups (F=39.78, P<0.05). The expressions of Col-Ⅰ mRNA were up-regulated by 1.29, 1.52, 2.11 fold, significant differences were observed in all groups (F=23.45, P<0.05); the expressions of protein were up-regulated by 1.18, 1.49, 2.11 fold and significant differences were observed in all groups (F=29.10, P<0.05). Compared with 0 hour group, the expressions of HSP47 mRNA were up-regulated by 1.56, 1.84, 2.86 fold in ARPE-19 cells stimulated by 5 ng/ml TGF-β2 for 12, 24 and 48 hours, and the differences were all significant (F=31.56, P<0.05); the expressions of protein were up-regulated by 2.08, 2.37, 2.80 fold, and the differences were all significant (F=49.18, P<0.05). The expressions of Col-Ⅰ mRNA were up-regulated by 1.57, 1.86, 2.78 fold and the differences were all significant (F=54.43, P<0.05), the expressions of protein were up-regulated by 1.38, 1.59, 2.16 fold and the differences were all significant (F=42.52, P<0.05). ConclusionTGF-β2 may play a role in the pathologic process of PVR by promoting the expression of HSP47 and then increasing the synthesis and accumulation of Col-Ⅰ.