The Role of Wnt/β-Catenin Signaling and K-Cadherin in the Regulation of Intraocular Pressure.
The Role of Wnt/β-Catenin Signaling and K-Cadherin in the Regulation of Intraocular Pressure.
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DOI：
10.1167/iovs.17-21964
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发表时间：
2018-03-01
影响因子：
4.4
通讯作者：
Clark AF
中科院分区：
文献类型：
--
作者：
Webber HC;Bermudez JY;Millar JC;Mao W;Clark AF
小梁网（TM）中的Wnt/β-catenin信号是维持正常眼压（IOP）所必需的，尽管其背后的机制尚不清楚。我们假设Wnt/β-catenin信号通过β-catenin对钙粘蛋白连接的影响来调节IOP。用100 ng/ml Wnt3a、1 μg/ml sFRP1或两者同时作用于非瘤性人TM （NTM）细胞4 ~ 48小时。对细胞进行β-连环蛋白、总钙粘蛋白或钙粘蛋白同种异型的免疫染色。分离膜蛋白或全细胞裂解物进行Western免疫印迹，并检测钙粘蛋白同型体。提取RNA合成cDNA， qPCR分析cadherin表达。部分NTM细胞在平板上培养进行细胞阻抗测定。Ad5。将编码K-cadherin和/或sFRP1的CMV重组腺病毒注射到4- 6月龄雌性BALB/cJ小鼠的眼睛中（n = 8-10）。连续35天评估有意识IOPs。经Wnt3a处理后，总cadherin表达增加，β-catenin在TM细胞膜和TM细胞间形成的突起上积累。qPCR结果显示，Wnt3a显著增加了NTM细胞中K-cadherin的表达（P < 0.01, n = 3）， Western免疫印迹检测显示，Wnt3a增加了NTM细胞中K-cadherin的表达，而sFRP1的加入抑制了这种表达。细胞阻抗分析显示，Wnt3a处理增加了跨细胞耐药性，抗k -cadherin siRNA降低了跨细胞耐药性（P < 0.001, n = 4-6）。我们的体内研究表明，K-cadherin可显著降低sfrp1诱导的高眼压（P < 0.05, n = 6）。Western免疫印迹也显示K-cadherin减轻了sfrp1诱导的小鼠前节段β-catenin的减少。我们的研究结果表明，钙粘蛋白通过Wnt/β-catenin通路在TM稳态和IOP调控中发挥重要作用。
Wnt/β-catenin signaling in the trabecular meshwork (TM) is required for maintaining normal intraocular pressure (IOP), although the mechanism(s) behind this are unknown. We hypothesize that Wnt/β-catenin signaling regulates IOP via β-catenin's effects on cadherin junctions. Nonglaucomatous primary human TM (NTM) cells were treated with or without 100 ng/ml Wnt3a, 1 μg/ml sFRP1, or both for 4 to 48 hours. Cells were immunostained for β-catenin, total cadherins, or cadherin isoforms. Membrane proteins or whole-cell lysates were isolated for Western immunoblotting and probed for cadherin isoforms. RNA was extracted for cDNA synthesis and qPCR analysis of cadherin expression. Some NTM cells were cultured on electric plates for cell impedance assays. Ad5.CMV recombinant adenoviruses encoding K-cadherin, and/or sFRP1 were injected into eyes of 4- to 6-month-old female BALB/cJ mice (n = 8–10). Conscious IOPs were assessed for 35 days. Upon Wnt3a treatment, total cadherin expression increased and β-catenin accumulated at the TM cell membrane and on processes formed between TM cells. qPCR showed that Wnt3a significantly increased K-cadherin expression in NTM cells (P < 0.01, n = 3), and Western immunoblotting showed that Wnt3a increased K-cadherin in NTM cells, which was inhibited by the addition of sFRP1. Cell impedance assays showed that Wnt3a treatment increased transcellular resistance and anti-K-cadherin siRNA decreased transcellular resistance (P < 0.001, n = 4–6). Our in vivo study showed that K-cadherin significantly decreased sFRP1-induced ocular hypertension (P < 0.05, n = 6). Western immunoblotting also showed that K-cadherin alleviated sFRP1-induced β-catenin decrease in mouse anterior segments. Our results suggest that cadherins play important roles in the regulation of TM homeostasis and IOP via the Wnt/β-catenin pathway.
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影响因子：
4.4
作者：
Filla MS;Schwinn MK;Sheibani N;Kaufman PL;Peters DM
通讯作者：
Peters DM
影响因子：
4.4
作者：
Mao, Weiming;Liu, Yang;Clark, Abbot F.
通讯作者：
Clark, Abbot F.
DOI：
10.1136/bjophthalmol-2015-307223
发表时间：
2016-01
期刊：
The British journal of ophthalmology
影响因子：
--
作者：
Kapetanakis VV;Chan MP;Foster PJ;Cook DG;Owen CG;Rudnicka AR
通讯作者：
Rudnicka AR
DOI：
10.1073/pnas.94.13.6770
发表时间：
1997-06-24
影响因子：
11.1
作者：
Finch, PW;He, X;Rubin, JS
通讯作者：
Rubin, JS
影响因子：
4.4
作者：
Mao, Weiming;Millar, J. Cameron;Clark, Abbot F.
通讯作者：
Clark, Abbot F.