吡非尼酮通过 PI3K/AKT 信号通路抑制成纤维细胞增殖、迁移或黏附，并减少大鼠硬膜外纤维化。
Pirfenidone inhibits fibroblast proliferation, migration or adhesion and reduces epidural fibrosis in rats via the PI3K/AKT signaling pathway.
机构信息
Dalian Medical University, Dalian, Liaoning, 116044, China.
Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, China.
出版信息
Biochem Biophys Res Commun. 2021 Apr 2;547:183-191. doi: 10.1016/j.bbrc.2021.01.055. Epub 2021 Feb 19.
OBJECTIVE
This present study aims to assess the effect of pirfenidone (PFD) on inhibiting fibroblast proliferation, migration or adhesion in vitro and reducing laminectomy-induced epidural fibrosis in vivo.
METHODS
The effect of PFD on proliferation inhibition was evaluated with flow cytometry, CCK-8, EdU and western-blotting assays. Altered properties in migration and adhesion were confirmed by wound-scratch, transwell, immunofluorescence (IF), cell adhesion and western-blotting assays. Additionally, fifty male healthy Sprague-Dawley rats were subjected to laminectomy and then treated with various concentrations of PFD. After 4 weeks, the degree of epidural fibrosis was evaluated by histological analysis.
RESULTS
In vitro, the results of flow cytometry, CCK-8, EdU and western-blotting assays showed that PFD reduced fibroblast proliferation by a dose-dependent manner. And the results of wound-scratch, transwell, IF, cell adhesion and western-blotting assays showed that the migration and adhesion of fibroblasts could be inhibited and the cytoskeleton could also be altered in a dose-dependent manner. And the inhibitory effect of PFD could be partially reversed in the PI3K overexpression experiment, which indicated that the capability of PFD to inhibit fibroblast proliferation, migration and adhesion might be through the PI3K/AKT signaling pathway. In vivo, an obvious reduction in epidural fibrosis was observed in groups topically treated with PFD.
CONCLUSIONS
Topical PFD application obviously suppressed laminectomy-induced epidural fibrosis, possibly by inhibiting fibroblast proliferation, migration and adhesion via the PI3K/AKT signaling pathway. PFD may be a safe and effective pharmaceutical to reduce clinical epidural fibrosis.
目的
本研究旨在评估吡非尼酮（PFD）对体外抑制成纤维细胞增殖、迁移或黏附的作用，并减少体内椎板切除术后硬膜外纤维化。
方法
通过流式细胞术、CCK-8、EdU 和 Western-blotting 测定评估 PFD 对增殖抑制的作用。通过划痕实验、Transwell 实验、免疫荧光（IF）、细胞黏附实验和 Western-blotting 实验证实迁移和黏附特性的改变。此外，50 只雄性健康 Sprague-Dawley 大鼠行椎板切除术，然后用不同浓度的 PFD 处理。4 周后，通过组织学分析评估硬膜外纤维化程度。
结果
在体外，流式细胞术、CCK-8、EdU 和 Western-blotting 实验结果表明，PFD 呈剂量依赖性降低成纤维细胞增殖。划痕实验、Transwell 实验、IF、细胞黏附实验和 Western-blotting 实验结果表明，PFD 可以抑制成纤维细胞的迁移和黏附，并且可以改变细胞骨架，呈剂量依赖性。在 PI3K 过表达实验中，PFD 的抑制作用部分逆转，表明 PFD 抑制成纤维细胞增殖、迁移和黏附的能力可能通过 PI3K/AKT 信号通路。在体内，局部应用 PFD 可明显减轻椎板切除术后硬膜外纤维化。
结论
局部应用 PFD 明显抑制椎板切除术后硬膜外纤维化，可能通过 PI3K/AKT 信号通路抑制成纤维细胞增殖、迁移和黏附。PFD 可能是一种安全有效的药物，可减少临床硬膜外纤维化。
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