Natural Plant Extract Berbamine Is a Potent Inhibitor of Cell Growth and Survival of Human Tenon’s Fibroblasts
Natural Plant Extract Berbamine Is a Potent Inhibitor of Cell Growth and Survival of Human Tenon’s Fibroblasts
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DOI：
10.1159/000506644
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发表时间：
2020-02
影响因子：
2.1
通讯作者：
Hui Xu;Yanan Kong;Ying Chen;Na Li;Shuqiang Zhang;Hong Lu
中科院分区：
文献类型：
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作者：
Hui Xu;Yanan Kong;Ying Chen;Na Li;Shuqiang Zhang;Hong Lu
简介：由于人特农成纤维细胞（HTF）过度增殖而导致的小梁切除术后疤痕常常导致手术失败。开发一种高效抑制HTF细胞生长的新型抗纤维化药物将大大提高小梁切除术的疗效。目的：本研究旨在探讨小檗胺（BBM）处理对 HTF 细胞生长和存活的影响。方法：用或不用不同浓度的 BBM 处理培养的人胎儿 Tenon 成纤维细胞 (HFTF)。用相差显微镜观察细胞形态。使用CCK-8方法和Ki67免疫荧光测定细胞活力和细胞增殖。使用划痕测试来研究细胞迁移。流式细胞术和TUNEL染色检测细胞凋亡。通过蛋白质印迹法测定 BAX/BCL-2、ERK 和 AKT/mTOR 通路成分的表达。结果：BBM 治疗破坏了 HFTF 的正常形态，并以剂量依赖性方式抑制其细胞生长。 Ki67 免疫荧光和划痕实验显示 BBM 抑制 HFTF 细胞增殖和迁移。重要的是，BBM 剂量依赖性地增加了 BAX/BCL-2 比率并诱导 HFTF 细胞凋亡。 Western blotting显示BBM显着抑制ERK和AKT/mTOR通路，PTEN抑制改善了BBM对HFTF细胞活力和存活的抑制作用。结论：BBM 通过 AKT/mTOR 有效抑制 HTF 的细胞生长和存活，具有作为小梁切除术后抗纤维化药物的潜力。
Introduction: Post-trabeculectomy scarring due to excessive proliferation of human Tenon’s fibroblasts (HTFs) often led to operation failure. Developing a new anti-fibrosis drug with high efficacy to inhibit HTF cell growth will greatly improve the effectiveness of trabeculectomy. Objective: This study aims to investigate the effect of berbamine (BBM) treatment on the cell growth and survival of HTFs. Methods: Cultured human fetal Tenon’s fibroblasts (HFTFs) were treated with or without different concentrations of BBM. Cell morphology was observed with a phase contrast microscope. A CCK-8 method and Ki67 immunofluorescence were used to determine cell viability and cell proliferation. A scratch test was used to study cell migration. Flow cytometry and TUNEL staining were performed to detect cell apoptosis. The expression of BAX/BCL-2, ERK, and AKT/mTOR pathway components was determined by Western blotting. Results: BBM treatment disrupted HFTF normal morphology and inhibited its cell growth in a dose-dependent manner. Ki67 immunofluorescence and scratch assay showed BBM suppressed HFTF cell proliferation and migration. Importantly, BBM dose-dependently increased the BAX/BCL-2 ratio and induced apoptosis in HFTF cells. Western blotting showed BBM significantly inhibited the ERK and AKT/mTOR pathway, and PTEN inhibition ameliorated the inhibitory effect of BBM on cell viability and survival in HFTFs. Conclusions: BBM potently inhibits the cell growth and survival of HTFs through AKT/mTOR and has the potential to serve as an anti-fibrosis drug after trabeculectomy.