热休克蛋白47在青光眼滤过泡瘢痕化中作用及其机制的研究

Glaucoma is a leading cause of irreversible blindness throughout the world. Extraocular filtering surgery is one of the main operations to treat patients with glaucoma. Successful glaucoma filtering procedures are characterized by the functional filtering blebs. Over-proliferation of Tenon capsule fibroblasts is a dominating reason resulting in scarring of the blebs, which is the most common cause of failure in glaucoma filtering surgery. Antimetabolites used clinically to prevent bleb scarring, however, serious complications happened. Heat shock protein 47 (HSP47) is closely related to fibrosis and scar formation. Inhibiting the expression of HSP47 could alleviate fibrosis. On the basis of our preliminary studies, we firstly proposed, both at home and abroad, that HSP47 may become a new candidate molecule of antifibrotic treatment in the filtering bleb scarring. This study is going to detect HSP47 levels in peripheral blood, aqueous humor, tears, and Tenon tissue of patients with glaucoma, and to analyse the role and its mechanisms of HSP47 in the process of glaucoma filtering bleb scarring. Construction of HSP47 specific dsRNA carrier, in vitro and in vivo studies will be finished in order to understand the effect and side reactions of siRNA targeting down HSP47 gene in inhibiting bleb scarring. A promising drug may be explored to improve the success rate of glaucoma surgery, and a new approach would be found to prevent bleb scarring after glaucoma filtration surgery.

青光眼是第一位不可逆性致盲眼病。眼外滤过术是治疗青光眼的主要手术，其成功与否与功能性滤过泡的形成相关。Tenon囊成纤维细胞过度增殖是导致滤过泡瘢痕化的主要原因，往往导致青光眼滤过术失败。虽然抗代谢药物已在临床用于抑制滤过泡瘢痕化，但会引起严重不良反应。热休克蛋白47( HSP47）与纤维化发生、发展和瘢痕形成密切相关,抑制HSP47可以减弱滤过泡纤维化程度。在前期研究基础之上，我们在国内外率先提出HSP47有可能成为防治青光眼滤过泡瘢痕化的新靶点。本项目通过检测青光眼患者外周血、眼房水、眼结膜囊泪液及瘢痕化滤过泡组织中HSP47的水平，分析HSP47在青光眼滤过泡瘢痕化中的作用及机制；构建HSP47特异性dsRNA载体进行离体实验和在体研究，了解siRNA靶向下调HSP47基因在抑制青光眼滤过泡瘢痕化中的效果和副作用，从而为防治青光眼滤过泡瘢痕化和提高青光眼滤过术的成功率探索新途径。

青光眼是全球第一位不可逆性致盲眼病。眼外滤过术是治疗青光眼的主要手术，其成功与否与功能性滤过泡的形成相关。Tenon囊成纤维细胞过度增殖是导致滤过泡瘢痕化的主要原因，往往导致青光眼滤过术失败。虽然抗代谢药物已在临床用于抑制滤过泡瘢痕化，但会引起严重不良反应。热休克蛋白47( HSP47）与纤维化发生、发展和瘢痕形成密切相关,抑制HSP47可以减弱滤过泡纤维化程度。我们在国内外率先提出HSP47有可能成为防治青光眼滤过泡瘢痕化的新靶点。通过研究发现，HSP47在青光眼患者外周血中的表达与正常对照组无差异，但在房水中的表达增高，并且在青光眼患者滤过泡瘢痕化组织中的表达较正常结膜组织增加。通过分离胎鼠眼部Tenon组织，成功培养了Tenon囊成纤维细胞，通过抑制Tenon囊成纤维细胞中HSP47的表达，成纤维细胞中胶原蛋白的表达水平下降。通过制作成年大鼠结膜滤过泡模型，并在结膜下注射HSP47 shRNA质粒，研究发现该质粒可抑制HSP47基因表达下调，结膜滤过泡中I型胶原和III型胶原蛋白的表达水平下降。形态学观察发现，结膜下注射HSP47 shRNA质粒可减轻滤过泡瘢痕化，这为防治青光眼滤过泡瘢痕化和提高青光眼滤过术的成功率探要索新途径。