Graduate Institute of Biomedical Sciences and Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Division of Chinese Internal Medicine, Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
Br J Pharmacol. 2021 Oct;178(20):4069-4084. doi: 10.1111/bph.15597. Epub 2021 Jul 14.
Neutrophil overactivation is crucial in the pathogenesis of acute lung injury (ALI). Bletinib (3,3'-dihydroxy-2',6'-bis(p-hydroxybenzyl)-5-methoxybibenzyl), a natural bibenzyl, extracted from the Bletilla plant, exhibits anti-inflammatory, antibacterial, and antimitotic effects. In this study, we evaluated the therapeutic effects of bletinib in human neutrophilic inflammation and LPS-mediated ALI in mice.
In human neutrophils activated with the formyl peptide (fMLP), we assessed integrin expression, superoxide anion production, degranulation, neutrophil extracellular trap (NET) formation, and adhesion through flow cytometry, spectrophotometry, and immunofluorescence microscopy. Immunoblotting was used to measure phosphorylation of Src family kinases (SFKs) and downstream proteins. Finally, a LPS-induced ALI model in male BALB/c mice was used to investigate the potential therapeutic effects of bletinib treatment.
In activated human neutrophils, bletinib reduced degranulation, respiratory burst, NET formation, adhesion, migration, and integrin expression; suppressed the enzymic activity of SFKs, including Src, Lyn, Fgr, and Hck; and inhibited the phosphorylation of SFKs as well as Vav and Bruton's tyrosine kinase (Btk). In mice with ALI, the pulmonary sections demonstrated considerable amelioration of prominent inflammatory changes, such as haemorrhage, pulmonary oedema, and neutrophil infiltration, after bletinib treatment.
Bletinib regulates neutrophilic inflammation by inhibiting the SFK-Btk-Vav pathway. Bletinib ameliorates LPS-induced ALI in mice. Further biochemical optimisation of bletinib may be a promising strategy for the development of novel therapeutic agents for inflammatory diseases.
中性粒细胞过度激活在急性肺损伤（ALI）的发病机制中至关重要。布替利珠（3,3'-二羟基-2',6'-双（对羟基苄基）-5-甲氧基联苯，一种从白芨植物中提取的天然联苯）具有抗炎、抗菌和抗有丝分裂作用。在这项研究中，我们评估了布替利珠在人类中性粒细胞炎症和 LPS 介导的小鼠 ALI 中的治疗作用。
在被甲酰肽（fMLP）激活的人中性粒细胞中，我们通过流式细胞术、分光光度法和免疫荧光显微镜评估整合素表达、超氧阴离子产生、脱颗粒、中性粒细胞胞外陷阱（NET）形成和黏附。免疫印迹用于测量 Src 家族激酶（SFK）和下游蛋白的磷酸化。最后，使用 LPS 诱导的 BALB/c 雄性小鼠 ALI 模型来研究布替利珠治疗的潜在治疗效果。
在激活的人中性粒细胞中，布替利珠减少脱颗粒、呼吸爆发、NET 形成、黏附、迁移和整合素表达；抑制 SFK 的酶活性，包括Src、Lyn、Fgr 和 Hck；并抑制 SFK 以及 Vav 和 Bruton 酪氨酸激酶（Btk）的磷酸化。在 ALI 小鼠中，布替利珠治疗后，肺切片显示明显改善了明显的炎症变化，如出血、肺水肿和中性粒细胞浸润。
布替利珠通过抑制 SFK-Btk-Vav 通路调节中性粒细胞炎症。布替利珠改善 LPS 诱导的小鼠 ALI。进一步对布替利珠进行生化优化可能是开发新型炎症性疾病治疗药物的有前途策略。
