Huashan Hospital, Department of Ophthalmology, Fudan University, Shanghai, China.
Invest Ophthalmol Vis Sci. 2015 Jan 6;56(2):855-64. doi: 10.1167/iovs.14-15301.
To examine the expression of Angiotensin II (Ang II) and its type I, and type II receptors (AT1R, AT2R) in rabbit Tenon's capsule fibroblasts after trabeculectomy, and to investigate the effects of Ang II on cultured human Tenon's capsule fibroblasts (HTFs) proliferation, migration, phenotype transition, and extracellular matrix (ECM) synthesis.
In the rabbit, expression of Ang II, AT1R, and AT2R in Tenon's capsule fibroblasts of eyes after trabeculectomy was evaluated by immunohistochemistry. Ang II levels in aqueous humor and plasma were assessed by ELISA. Cultured HTFs, obtained from patients undergoing cataract surgery, were treated with Ang II, TGF-β1, or vehicle control. Cell proliferation and migration were evaluated by Cell Counting Kit-8 and Transwell assay, and wound scratch assay, respectively. Protein expressions of α-smooth muscle actin (α-SMA) and fibronectin (FN) were measured by Western blot and immunofluorescence. Messenger RNA expressions of α-SMA and FN were measured by real-time PCR.
In the rabbit, the expression of Ang II and AT1R increased from 1 day after surgery while AT2R increased from 7 days. In cultured HTFs, Ang II promoted cell proliferation and migration significantly (P < 0.05). Interestingly, the effect of 10(-7) M Ang II was more prominent than higher concentrations (10(-5) M; P < 0.05). Ang II also markedly induced the expression of α-SMA and FN, suggesting a phenotypic transition to myofibroblasts.
Our results show that trabeculectomy alter the levels of Ang II and its receptors in Tenon's capsule fibroblasts, and that Ang II increase HTFs proliferation, migration, and phenotype transition, suggesting that Ang II may play a role in wound healing after trabeculectomy.
研究兔眼小梁切除术后腱膜囊成纤维细胞中血管紧张素 II（Ang II）及其 I 型和 II 型受体（AT1R、AT2R）的表达情况，并探讨 Ang II 对培养的人腱膜囊成纤维细胞（HTFs）增殖、迁移、表型转化和细胞外基质（ECM）合成的影响。
采用免疫组织化学方法检测兔小梁切除术后腱膜囊成纤维细胞中 Ang II、AT1R 和 AT2R 的表达。通过 ELISA 检测房水和血浆中 Ang II 水平。从接受白内障手术的患者中获取培养的 HTFs，用 Ang II、TGF-β1 或载体对照处理。通过细胞计数试剂盒-8 和 Transwell 测定分别评估细胞增殖和迁移，通过划痕实验评估细胞迁移。通过 Western blot 和免疫荧光法测定α-平滑肌肌动蛋白（α-SMA）和纤维连接蛋白（FN）的蛋白表达。通过实时 PCR 测定 α-SMA 和 FN 的信使 RNA 表达。
在兔模型中，Ang II 和 AT1R 的表达从术后第 1 天开始增加，而 AT2R 的表达从术后第 7 天开始增加。在培养的 HTFs 中，Ang II 显著促进细胞增殖和迁移（P<0.05）。有趣的是，10(-7)M Ang II 的作用比高浓度（10(-5)M；P<0.05）更显著。Ang II 还显著诱导了 α-SMA 和 FN 的表达，提示成纤维细胞向肌成纤维细胞的表型转化。
我们的结果表明，小梁切除术改变了腱膜囊成纤维细胞中 Ang II 及其受体的水平，Ang II 增加了 HTFs 的增殖、迁移和表型转化，提示 Ang II 可能在小梁切除术后的伤口愈合中发挥作用。
