Song Jia, Zhang Jiancheng, Shi Jun, Pan Xuming, Mo Dayu
Department of General Practice, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People's Republic of China.
Department of Emergency, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People's Republic of China.
J Inflamm Res. 2024 Aug 1;17:5161-5176. doi: 10.2147/JIR.S446345. eCollection 2024.
Breviscapine has been demonstrated to have beneficial effects in ameliorating acute lung injury (ALI), yet its potential therapeutic value and molecular mechanisms in sepsis-induced ALI remain unexplored.
We utilized network pharmacology approach to identify the potential targets and mechanisms of breviscapine in treating sepsis-induced ALI. To construct a murine model of sepsis, we performed cecal ligation and puncture (CLP). Hematoxylin and eosin (HE) staining and enzyme-linked immunosorbent assay (ELISA) were employed to respectively determine the pathologic changes and levels of inflammatory factors. Neutrophil count and total protein level in bronchoalveolar lavage fluid (BALF) were detected by corresponding kit. Additionally, we utilized flow cytometry, immunofluorescence, Western blotting, and real-time reverse transcription PCR (qRT-PCR) to detect cell apoptosis, protein expression, and gene expression. Finally, we used ELISA kits to detect the activity of myeloperoxidase (MPO) and caspase-8 (CASP8).
Breviscapine was revealed to target 81 potential proteins in the treatment of sepsis-induced ALI, while CASP8 was the most important one as demonstrated by network analysis. In vivo experiments demonstrated that breviscapine effectively reduced the severity of sepsis-induced ALI and inflammation, and significantly suppressed neutrophil infiltration in the lung tissues of CLP mice and promoted neutrophil apoptosis in the peripheral blood. In vitro experiments revealed that lipopolysaccharide (LPS)-induced neutrophil apoptosis was inhibited, and the expression and activity of CASP8 were down-regulated. Breviscapine intervention markedly up-regulated the expression and activity of CASP8, consequently activating neutrophil apoptosis and inhibiting inflammatory response by activating the NF-κB signaling pathway.
Breviscapine is remarkably effective in improving sepsis-induced ALI, and its mechanism of action may be to induce neutrophil apoptosis, inhibit inflammatory overreaction and reduce its infiltration in pulmonary tissues by up-regulating the expression and activity of CASP8.
灯盏花素已被证明在改善急性肺损伤（ALI）方面具有有益作用，但其在脓毒症诱导的ALI中的潜在治疗价值和分子机制仍未被探索。
我们利用网络药理学方法来确定灯盏花素治疗脓毒症诱导的ALI的潜在靶点和机制。为构建脓毒症小鼠模型，我们进行了盲肠结扎和穿刺（CLP）。采用苏木精-伊红（HE）染色和酶联免疫吸附测定（ELISA）分别确定病理变化和炎症因子水平。通过相应试剂盒检测支气管肺泡灌洗液（BALF）中的中性粒细胞计数和总蛋白水平。此外，我们利用流式细胞术、免疫荧光、蛋白质印迹和实时逆转录PCR（qRT-PCR）检测细胞凋亡、蛋白质表达和基因表达。最后，我们使用ELISA试剂盒检测髓过氧化物酶（MPO）和半胱天冬酶-8（CASP8）的活性。
网络分析表明，灯盏花素在治疗脓毒症诱导的ALI中靶向81种潜在蛋白质，而CASP8是最重要的一种。体内实验表明，灯盏花素有效降低了脓毒症诱导的ALI的严重程度和炎症反应，并显著抑制CLP小鼠肺组织中的中性粒细胞浸润，促进外周血中性粒细胞凋亡。体外实验表明，脂多糖（LPS）诱导的中性粒细胞凋亡受到抑制，CASP8的表达和活性下调。灯盏花素干预显著上调CASP8的表达和活性，从而通过激活NF-κB信号通路激活中性粒细胞凋亡并抑制炎症反应。
灯盏花素在改善脓毒症诱导的ALI方面非常有效，其作用机制可能是通过上调CASP8的表达和活性来诱导中性粒细胞凋亡，抑制炎症过度反应并减少其在肺组织中的浸润。
