Fu Shuhao, Lu Zhaozeng, Ye Wen
Department of Ophthalmology, Huashan Hospital, Fudan University, Shanghai 200040, China.
Evid Based Complement Alternat Med. 2022 Jul 12;2022:7729827. doi: 10.1155/2022/7729827. eCollection 2022.
To assess Interlukin-11 (IL11) expression in the tears of patients after filtration surgery and to find out its pro-transdifferentiational and pro-fibrotic functions and mechanisms on subconjunctival human Tenon's capsule fibroblasts (HTFs) induced by transforming growth factor beta1 (TGF-1).
Tears were collected from glaucoma patients with or without filtration surgery. The expression of IL11 in tears was examined by enzyme-linked immunosorbent assay. Primary HTFs were prepared as an expansion culture of human Tenon's explants from patients undergoing cataract surgery. TGF-1 and IL11 were used to stimulate the cultured HTFs. Quantitative RT-PCR and western blot analyzed the roles of TGF-1 in IL11 and DNA methyltransferase (DNMT) expression and the effects of IL11 on collagen-1A1 and -smooth muscle actin expression. The effects of IL11 on human HTFs' migration were tested via the scratch-wound assay. MassARRAY platform of Sequenom was applied for analyzing the quantitative methylation of the IL11 promoter region.
Our data presented significantly high levels of IL11 in the tears of patients who underwent filtration surgery with uncontrolled intraocular pressure (IOP) compared with those who underwent filtration surgery with controlled IOP. The up-regulation of IL11 was related to TGF-1. We also found that TGF- induced IL11 up-regulation in the HTFs, which activates the HTFs and enhanced the translation of the pro-fibrotic protein expression. This is correlated with inhibiting the activity and expression of DNMTs and demethylating the IL11 promoter. Therefore, IL11 may be an ideal target to be regulated to control the filtering pathway scar formation.
评估滤过性手术后患者泪液中白细胞介素-11（IL11）的表达，并探究其对转化生长因子β1（TGF-1）诱导的人结膜下Tenon囊成纤维细胞（HTFs）的促转分化和促纤维化功能及机制。
收集有或无滤过性手术的青光眼患者的泪液。采用酶联免疫吸附测定法检测泪液中IL11的表达。从白内障手术患者的人Tenon囊外植体中制备原代HTFs并进行扩增培养。用TGF-1和IL11刺激培养的HTFs。定量逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法分析TGF-1对IL11和DNA甲基转移酶（DNMT）表达的作用，以及IL11对Ⅰ型胶原α1（collagen-1A1）和α-平滑肌肌动蛋白表达的影响。通过划痕试验检测IL11对人HTFs迁移的影响。应用Sequenom公司的MassARRAY平台分析IL11启动子区域的定量甲基化。
我们的数据显示，与眼压得到控制的滤过性手术患者相比，眼压未得到控制的滤过性手术患者泪液中IL11水平显著升高。IL11上调与TGF-1有关。我们还发现，TGF-1可诱导HTFs中IL-11上调，激活HTFs并增强促纤维化蛋白表达的翻译。这与抑制DNMTs的活性和表达以及使IL11启动子去甲基化相关。因此，IL11可能是调控以控制滤过通道瘢痕形成的理想靶点。
