阿米替林治疗百草枯诱导的肺纤维化的潜力：涉及窖蛋白-1 介导的抗上皮-间充质转化和抑制细胞凋亡。
Therapeutic potential of amitriptyline for paraquat-induced pulmonary fibrosis: Involvement of caveolin-1-mediated anti-epithelial-mesenchymal transition and inhibition of apoptosis.
机构信息
Department of Intensive Care Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Department of Poisoning and Occupational Diseases, Qilu Hospital of Shandong University, Jinan 250000, China.
出版信息
Ecotoxicol Environ Saf. 2023 Apr 1;254:114732. doi: 10.1016/j.ecoenv.2023.114732. Epub 2023 Mar 8.
OBJECTIVE
Treatment of pulmonary fibrosis caused by paraquat (PQ) poisoning remains problematic. Amitriptyline (AMT) has multiple pharmacological effects. Here we investigated the anti-fibrotic effect of AMT on PQ-induced pulmonary fibrosis and its possible mechanism.
METHODS
C57BL/6 mice were randomly divided into control, PQ, PQ + AMT and AMT groups. Histopathology of the lungs, blood gas analysis, and levels of hydroxyproline (HYP), transforming growth factor β1 (TGF-β1) and interleukin 17 (IL-17) were measured. The siRNA transfection inhibited caveolin-1 in A549 cells, which induced epithelial-mesenchymal transition (EMT) by PQ and followed intervention with AMT. E-cadherin, N-cadherin, α-smooth muscle actin (α-SMA) and caveolin-1 were studied by immunohistochemistry and western blot analysis. The apoptosis rate was measured by flow cytometry.
RESULTS
Compared with the PQ group, the PQ + AMT group displayed mild pathological changes in pulmonary fibrosis, lower HYP, IL-17 and TGF- β1 levels in lung, but high TGF- β1 in serum. Levels of N-cadherin and α-SMA in the lungs were significantly decreased, but caveolin-1 was increased, while SaO and PaO levels were higher. Compared with the PQ group, the apoptosis rate, N-cadherin and α-SMA levels in A549 cells were significantly decreased after PQ treatment and high dose AMT intervention (p < 0.01). The expressions of E-cadherin, N-cadherin and α-SMA in the PQ-induced cells transfected with caveolin-1 siRNA or siControl RNA were significantly different (p < 0.01), but the apoptosis rate was unaltered.
CONCLUSION
AMT inhibited PQ-induced EMT in A549 cells and improved lung histopathology and oxygenation in mice by up-regulating caveolin-1.
目的
百草枯（PQ）中毒引起的肺纤维化的治疗仍然存在问题。阿米替林（AMT）具有多种药理作用。在这里，我们研究了 AMT 对 PQ 诱导的肺纤维化的抗纤维化作用及其可能的机制。
方法
将 C57BL/6 小鼠随机分为对照组、PQ 组、PQ+AMT 组和 AMT 组。测量肺组织病理学、血气分析以及羟脯氨酸（HYP）、转化生长因子β1（TGF-β1）和白细胞介素 17（IL-17）的水平。通过 PQ 诱导 A549 细胞上皮-间充质转化（EMT），并随后用 AMT 进行干预，用 siRNA 转染抑制小窝蛋白-1（caveolin-1）。通过免疫组织化学和 Western blot 分析研究 E-钙粘蛋白、N-钙粘蛋白、α-平滑肌肌动蛋白（α-SMA）和小窝蛋白-1。通过流式细胞术测量细胞凋亡率。
结果
与 PQ 组相比，PQ+AMT 组肺纤维化的病理变化较轻，肺组织 HYP、IL-17 和 TGF-β1 水平较低，但血清 TGF-β1 水平较高。肺组织中 N-钙粘蛋白和 α-SMA 水平明显降低，而 caveolin-1 升高，SaO 和 PaO 水平升高。与 PQ 组相比，PQ 处理和高剂量 AMT 干预后 A549 细胞的凋亡率、N-钙粘蛋白和 α-SMA 水平明显降低（p<0.01）。转染 caveolin-1 siRNA 或 siControl RNA 的 PQ 诱导细胞中 E-钙粘蛋白、N-钙粘蛋白和 α-SMA 的表达有明显差异（p<0.01），但细胞凋亡率无变化。
结论
AMT 通过上调小窝蛋白-1 抑制 PQ 诱导的 A549 细胞 EMT，并改善小鼠的肺组织病理学和氧合作用。
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