Huang Li-Na, Qiu Zheng-Ying, Pan Xiang-Yi, Liu Chen, Li Si-Fan, Ge Shao-Guang, Shi Xiong-Wei, Cao Hao, Xin Rui-Hua, Hu Fang-di
School of Pharmacy,Lanzhou University Lanzhou 730000,China.
Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences Lanzhou 730050,China.
Zhongguo Zhong Yao Za Zhi. 2025 Jun;50(11):3097-3107. doi: 10.19540/j.cnki.cjcmm.20250122.401.
Based on the interleukin-17(IL-17) signaling pathway, this study explores the effect and mechanism of Bufei Decoction on Klebsiella pneumoniae pneumonia in rats. SD rats were randomly divided into the control group, model group, Bufei Decoction low-dose group(6.68 g·kg(-1)·d(-1)), Bufei Decoction high-dose group(13.36 g·kg(-1)·d(-1)), and dexamethasone group(1.04 mg·kg(-1)·d(-1)), with 10 rats in each group. A pneumonia model was established by tracheal drip injection of K. pneumoniae. After successful model establishment, the improvement in lung tissue damage was observed following drug administration. Core targets and signaling pathways were screened using transcriptomics techniques. Real-time fluorescence quantitative polymerase chain reaction was used to detect the mRNA expression of core targets interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and chemokine CXC ligand 6(CXCL6). Western blot was used to assess key proteins in the IL-17 signaling pathway, including interleukin-17A(IL-17A), nuclear transcription factor-κB activator 1(Act1), tumor necrosis factor receptor-associated factor 6(TRAF6), and downstream phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK), and phosphorylated nuclear factor-κB p65(p-NF-κB p65). Apoptosis of lung tissue cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL). The results showed that, compared with the control group, the model group exhibited significant pathological damage in lung tissue. The mRNA expression of IL-6, IL-1β, TNF-α, and CXCL6, as well as the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly increased, and the number of apoptotic cells was notably higher, indicating successful model establishment. Compared with the model group, both low-and high-dose groups of Bufei Decoction showed reduced pathological damage in lung tissue. The mRNA expression levels of IL-6, IL-1β, TNF-α, and CXCL6, and the protein levels of IL-17A, Act1, TRAF6, p-p38 MAPK/p38 MAPK, and p-NF-κB p65/NF-κB p65, were significantly decreased, with a significant reduction in apoptotic cells in the high-dose group. In conclusion, Bufei Decoction can effectively improve lung tissue damage and reduce inflammation in rats with K. pneumoniae. The mechanism may involve the regulation of the IL-17 signaling pathway and the reduction of apoptosis.
本研究基于白细胞介素-17(IL-17)信号通路,探讨补肺汤对大鼠肺炎克雷伯菌肺炎的影响及作用机制。将SD大鼠随机分为对照组、模型组、补肺汤低剂量组(6.68 g·kg⁻¹·d⁻¹)、补肺汤高剂量组(13.36 g·kg⁻¹·d⁻¹)和地塞米松组(1.04 mg·kg⁻¹·d⁻¹),每组10只。通过气管滴注肺炎克雷伯菌建立肺炎模型。模型建立成功后,观察给药后肺组织损伤的改善情况。采用转录组学技术筛选核心靶点和信号通路。运用实时荧光定量聚合酶链反应检测核心靶点白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和趋化因子CXC配体6(CXCL6)的mRNA表达。采用蛋白质印迹法评估IL-17信号通路中的关键蛋白,包括白细胞介素-17A(IL-17A)、核转录因子-κB激活因子1(Act1)、肿瘤坏死因子受体相关因子6(TRAF6)以及下游磷酸化的p38丝裂原活化蛋白激酶(p-p38 MAPK)和磷酸化的核因子-κB p65(p-NF-κB p65)。通过末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法(TUNEL)检测肺组织细胞凋亡情况。结果显示,与对照组相比,模型组肺组织出现明显病理损伤。IL-6、IL-1β、TNF-α和CXCL6的mRNA表达以及IL-17A、Act1、TRAF6、p-p38 MAPK/p38 MAPK和p-NF-κB p65/NF-κB p65的蛋白水平显著升高,凋亡细胞数量明显增多,表明模型建立成功。与模型组相比,补肺汤低剂量组和高剂量组肺组织病理损伤均减轻。IL-6、IL-1β、TNF-α和CXCL6的mRNA表达水平以及IL-17A、Act1、TRAF6、p-p38 MAPK/p38 MAPK和p-NF-κB p65/NF-κB p65的蛋白水平均显著降低,高剂量组凋亡细胞显著减少。综上所述,补肺汤可有效改善肺炎克雷伯菌感染大鼠的肺组织损伤并减轻炎症反应。其机制可能涉及对IL-17信号通路的调节及凋亡的减少。