Arsenic Trioxide Inhibits Proliferation of Rabbit Tenon's Capsule Fibroblasts After Trabeculectomy by Downregulating Expression of Extracellular Matrix Proteins.
Arsenic Trioxide Inhibits Proliferation of Rabbit Tenon's Capsule Fibroblasts After Trabeculectomy by Downregulating Expression of Extracellular Matrix Proteins.
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DOI：
10.1167/iovs.15-17289
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发表时间：
2015-10
影响因子：
4.4
通讯作者：
Ying Su;Chenggong Jiang;Ling Zhang;Feng Wang
中科院分区：
文献类型：
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作者：
Ying Su;Chenggong Jiang;Ling Zhang;Feng Wang
目的研究三氧化二砷（ATO）对小梁切除术的调节作用。方法采用随机、对照、前瞻性研究设计。原代培养Tenon囊成纤维细胞（TF），用3-（4，5-二甲基噻唑基-2-）-2，5-二苯基四氮唑溴化盐（MTT）法和溴脱氧尿嘧啶核苷（BrdU）掺入法检测ATO对TF活性的影响。通过50%生长抑制（IC 50值）和细胞周期分析确定ATO的剂量。Western blotting检测细胞外基质（ECM）和增殖细胞核抗原（PCNA）的表达。HE染色观察滤过泡状态。结果MTT法和BrdU法显示，6 μM三氧化二砷作用于兔肾成纤维细胞（rTF）72 h后，其IC 50值明显升高。此外，ATO处理不仅导致ECM蛋白、纤连蛋白、IV型胶原和层粘连蛋白在rTF中的表达显著降低，而且还以时间依赖性方式降低PCNA表达。组织学观察显示溶剂组和对照组rTF均呈高水平增殖，实验组rTF增殖能力低于溶剂组和对照组。实验组小梁切除术后14 d未见明显炎症反应，出现滤过泡。结论三氧化二砷可抑制小梁切除术后rTFs的增殖，提高小梁切除术的成功率。
PURPOSE The purpose of this study was to study the effectiveness of arsenic trioxide (ATO) in regulation of trabeculectomy in an animal model. METHODS This study was designed as random, comparative, and prospective study. Primary Tenon's capsule fibroblasts (TFs) were cultured and their viability after arsenic trioxide (ATO) treatment was detected using 3-(4, 5-dimethylthiazolyl-2-)-2, 5-diphenyltetrazoliumbromide (MTT) and bromodeoxyuridine incorporation (BrdU) assays. The dose of ATO was determined by 50% growth inhibition (IC50 values) and cell-cycle analysis. The expression of extracellular matrix (ECM) components and proliferating cell nuclear antigen (PCNA) was determined by Western blotting analysis. The status of filtration bleb was detected by hematoxylin and eosins (HE) stain. RESULTS Based on the MTT and BrdU assays, IC50 values was observed in rabbit TFs (rTFs) cells after treatment with ATO at a concentration of 6 μM arsenic trioxide for 72 hours. In addition, the ATO treatment caused not only a significant reduction in the expression of ECM proteins, fibronectin, collagen IV, and laminin in rTFs, but also reduced the expression of PCNA expression in a time dependent manner. The histologic observation showed high-level proliferation of rTFs in both vehicle and control groups, and proliferative ability of rTFs of experimental group was less than in vehicle and control group. Moreover, no inflammation was observed in the experimental group, and the filtering bleb was detected in the experimental group after 14 days of trabeculectomy. CONCLUSIONS Arsenic trioxide inhibited the proliferation of rTFs after trabeculectomy and may improve the success ratio of trabeculectomy in an animal model.