Medical College, Qingdao University, Qingdao, China.
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Eye Institute of Shandong First Medical University, 5 Yan'erdao Road, Qingdao, 266071, China.
BMC Ophthalmol. 2021 Dec 5;21(1):419. doi: 10.1186/s12886-021-02186-w.
Topical application of β-blocker eye drops induces damage to the ocular surface in clinical. However, the mechanism involved remains incompletely understood. The purpose of this study was to investigate the influence and mechanism of β-blocker eye drops on corneal epithelial wound healing.
Corneal epithelial wound healing models were constructed by epithelial scraping including in the limbal region and unceasingly received eye drops containing 5 mg/mL β-blocker levobunolol, β1-adrenoceptor (β1AR)-specific antagonist atenolol or β2-adrenoceptor (β2AR)-specific antagonist ICI 118, 551. For the migration assay, the murine corneal epithelial stem/progenitor cells (TKE2) were wounded and subsequently incubated with levobunolol, atenolol, or ICI 118, 551. The proliferation and colony formation abilities of TKE2 cells treated with levobunolol, atenolol, or ICI 118, 551 were investigated by CCK-8 kit and crystal violet staining. The differentiation marker Cytokeratin 3 (CK3), the stem cell markers-Cytokeratin 14 (CK14) and Cytokeratin 19 (CK19), and corneal epithelium regeneration-related signaling including in Ki67 and the phosphorylated epithelial growth factor receptor (pEGFR) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) were assessed by immunofluorescence staining.
Levobunolol and ICI 118, 551 impaired corneal wound healing, decreased the expressions of CK3, CK14, and CK19 after limbal region scraping in vivo and reduced the migration and proliferation of TKE2 in vitro, whereas atenolol had no significant effect. Moreover, levobunolol and ICI 118, 551 inhibited corneal wound healing by mediating the expression of Ki67, and the phosphorylation of EGFR and ERK1/2 in the limbal and regenerated corneal epithelium.
β-blocker eye drops impaired corneal wound healing by inhibiting the β2AR of limbal stem cells, which decreased corneal epithelial regeneration-related signaling. Therefore, a selective β1AR antagonist might be a good choice for glaucoma treatment to avoid ocular surface damage.
局部应用β受体阻滞剂滴眼可导致临床眼表损伤。然而，其涉及的机制尚不完全清楚。本研究旨在探讨β受体阻滞剂滴眼对角膜上皮伤口愈合的影响及其机制。
通过包括在角膜缘区的上皮刮除构建角膜上皮伤口愈合模型，并持续滴注含有 5mg/ml 的β受体阻滞剂左布诺洛尔、β1 肾上腺素能受体（β1AR）特异性拮抗剂阿替洛尔或β2 肾上腺素能受体（β2AR）特异性拮抗剂 ICI 118,551 的眼药水。对于迁移实验，将鼠角膜上皮干细胞/祖细胞（TKE2）刮伤，然后用左布诺洛尔、阿替洛尔或 ICI 118,551 孵育。通过 CCK-8 试剂盒和结晶紫染色法研究 TKE2 细胞经左布诺洛尔、阿替洛尔或 ICI 118,551 处理后的增殖和集落形成能力。通过免疫荧光染色评估角膜上皮再生相关信号包括 Ki67 和磷酸化表皮生长因子受体（pEGFR）和磷酸化细胞外信号调节激酶 1/2（pERK1/2）以及分化标志物细胞角蛋白 3（CK3）、干细胞标志物细胞角蛋白 14（CK14）和细胞角蛋白 19（CK19）的表达。
左布诺洛尔和 ICI 118,551 损害角膜伤口愈合，体内角膜缘刮除后降低 CK3、CK14 和 CK19 的表达，并减少 TKE2 的体外迁移和增殖，而阿替洛尔则无明显作用。此外，左布诺洛尔和 ICI 118,551 通过调节 Ki67 的表达以及角膜缘和再生角膜上皮中的 EGFR 和 ERK1/2 的磷酸化来抑制角膜伤口愈合。
β受体阻滞剂滴眼通过抑制角膜缘干细胞的β2AR 抑制角膜上皮再生相关信号，从而损害角膜伤口愈合。因此，选择性β1AR 拮抗剂可能是治疗青光眼以避免眼表损伤的良好选择。
