Expression of hot shock protein 47 in pre-retinal membrane of proliferative vitreoretinopathy and the influence of transforming growth factor-β<sub>2</sub> on expression of hot shock protein 47 in retinal pigment epithelial cell_Chinese Journal of Ocular Fundus Diseases

Authors：

YuanLili , ZhangDongchang ,  LiXiaorong , 卢亚楠

Shanxi Eye Hospital, Taiyuan 030002, China;

Corresponding author：

LiXiaorong, Email: Xiaorli@163.com

Keywords：

Vitreoretinopathy, Proliferative/physiopathology; Retinal pigment epithelium; HSP47 heat-shock proteins; Transforming growth factor beta2

DOI：

10.3760/cma.j.issn.1005-1015.2015.04.016

Video：

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Objective To 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. Methods Pre-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. Results A 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). Conclusion TGF-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.

Citation： YuanLili, ZhangDongchang, LiXiaorong. Expression of hot shock protein 47 in pre-retinal membrane of proliferative vitreoretinopathy and the influence of transforming growth factor-β₂ on expression of hot shock protein 47 in retinal pigment epithelial cell. Chinese Journal of Ocular Fundus Diseases, 2015, 31(4): 377-381. doi: 10.3760/cma.j.issn.1005-1015.2015.04.016 Copy

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1. Ryan SJ.视网膜(第3卷)[M].黎晓新,赵家良,译.天津:天津科技翻译出版公司,2011: 2209-2211.
2. Mony S,Lee SJ,Harper JF,et al.Regulation of Na,K-ATPase β1-subunit in TGF-β₂-mediated epithelial-to-mesenchymal transition in human retinal pigmented epithelial cells[J].Exp Eye Res,2013,115:113-122.
3. Kakugawa T,Yokota S,Ishimatsu Y,et al.Serum heat shock protain 47 levels are elevated in idiopathic pulmonary fibrosis[J].Cell Stress Chaperones,2013,18(5): 581-590.
4. Kitamura H,Yamamoto S,Nakase H,et al.Role of heat shock protain in indestinal fibrosis of experimental colitis[J].Biochem Biophys Res Commun,2011,404(2):599-604.
5. Xiao HB,Liu RH,Ling GH,et al.HSP47 regulates ECM accumulation in renal proximal tubular cells induced by TGF-β1 through ERK1/2 and JNK MAPK pathyways[J].Am J Physiol Renal Physiol,2012,303(5):757-765.
6. 吕明良,李敏.转化生长因子β1对人视网膜色素上皮细胞热休克蛋白47及血管内皮生长因子表达的影响[J].中华眼底病杂志,2010,26(6):573-574.
7. Lee J,Ko M,Joo CK.Rho plays a key role in TGF-beta induced cytoskeletal rearrangement in human retinal pigment epithelium[J].J Cell Physiol,2008,216(2): 520-526.
8. Walsh MF,Ampasala DR,Hatfield J,et al.Transforming growth factor-β stimulates intestinal epithelial focal adhesion kinase synthesis via smad-and p38-dependent mechanisms[J].Am J Pathol,2008,173(2):385-399.
9. Rubtsov YP,Rudensky AY.TGF beta signalling in control of T-cell-mediated self-reactivity[J].Nat Rev Immunol,2007,7(6):443-453.
10. Taguchi T, Nazneen A,Al-Shihri AA,et al.Heat shock protain 47:a novel biomarker of phenotypically altered collagen-producing cells[J].Acta Histochem Cytochem, 2011,44(2):35-41.
11. Sauk JJ,Nikitakis N,Siavash H.Hsp47 a novel collagen binding serpin chaperone,autoantigen and therapeutic target[J].Front Biosci,2005,10:107-118.
12. Sandbo N,Lau A,Kach J,et al.Delayed stress fiber formation mediates pulmonary myofibroblast differentiation in response to TGF-β[J].Am J Physiol Lung Cell Mol Physiol,2011,301(5):656-666.
13. Philips N,Samuel P,Parakandi H,et al.Beneficial regulation of fibrillar collagens, heat shock protein-47, elastin fiber components, transforming growth factor-β1, vascular endothelial growth factor and oxidative stress effects by copper in dermal fibroblasts[J].Connect Tissue Res,2012,53(5):373-378.
14. Wang Z,Li L.Adenovirus-mediated RNA interference against collagen-specific molecular chaperone 47-KDa heat shock protein suppresses scar formation on mouse wounds[J].Cell Biol Int,2008,32(5):484-493.

Chinese Journal of Ocular Fundus Diseases

Expression of hot shock protein 47 in pre-retinal membrane of proliferative vitreoretinopathy and the influence of transforming growth factor-β₂ on expression of hot shock protein 47 in retinal pigment epithelial cell

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Chinese Journal of Ocular Fundus Diseases

Expression of hot shock protein 47 in pre-retinal membrane of proliferative vitreoretinopathy and the influence of transforming growth factor-β2 on expression of hot shock protein 47 in retinal pigment epithelial cell

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Expression of hot shock protein 47 in pre-retinal membrane of proliferative vitreoretinopathy and the influence of transforming growth factor-β₂ on expression of hot shock protein 47 in retinal pigment epithelial cell