马钱草颗粒通过调节NLRP3/p38 MAPK/甘油磷脂通路缓解吸烟合并肺炎链球菌引起的急性支气管炎

Wanxia Men, Ruipeng Liu, Min Yang, Yang Xu, Yanli Lei, Tao Liu
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The mRNA expression levels of NOD-like receptor thermal protein domain associated protein 3 ( NLRP3) and p38 mitogen-activated protein kinase ( p38 MAPK) in lung tissue were detected by real-time fluorescence quantitative polymerase chain reaction, and the expression levels of inflammatory factors (tumor necrosis factor-α [ TNF-α], interleukin-6 [ IL-6], interleukin-1β [ IL-1β], interleukin-10 [ IL-10]) in serum and lung homogenate were detected by enzyme-linked immunosorbent assay. The therapeutic effect of MQ on rats with AB was preliminarily evaluated. Metabolic profiling of rat serum was performed by ultra high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry/mass spectrometry, combining P < .05, variable important in projection > 1, fold change > 1.5 and screening for differential metabolites based on univariate and multivariate statistical analyses, and then analyzing the possible metabolic pathways with the help of MetaboAnalyst 5.0 database, to analyze the mechanism of MQ’s action on the treatment of AB at the metabolic level. Results: MQ significantly ameliorated the pathological injury of bronchial and lung tissues in model rats, reduced the mRNA levels of NLRP3 and p38 MAPK ( P < .05, .01), lowered the levels of TNF-α, IL-6, and IL-1β in serum and lung homogenates ( P < .05, .01), and elevated IL-10 in serum and lung homogenates ( P < .0001, .01) levels, and to regulate abnormal metabolites in rats with AB via the glycerophospholipid metabolic pathway. 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引用次数: 0

摘要

目的探讨马钱小儿颗粒(MQs)治疗急性支气管炎(AB)的疗效和作用机制。方法通过混合烟雾和肺炎链球菌鼻腔感染建立大鼠急性支气管炎模型。通过苏木精-伊红(HE)染色观察肺组织和支气管的病理变化并评分。实时荧光定量聚合酶链反应检测肺组织中 NOD 样受体热蛋白结构域相关蛋白 3(NLRP3)和 p38 丝裂原活化蛋白激酶(p38 MAPK)的 mRNA 表达水平、用酶联免疫吸附试验检测血清和肺匀浆中炎症因子(肿瘤坏死因子-α[TNF-α]、白细胞介素-6[IL-6]、白细胞介素-1β[IL-1β]、白细胞介素-10[IL-10])的表达水平。初步评估了 MQ 对 AB 大鼠的治疗效果。采用超高效液相色谱-四极杆-飞行时间质谱/质谱联用技术对大鼠血清进行代谢谱分析,结合P 1、折合变化>1.5和基于单变量和多变量统计分析筛选差异代谢物,然后借助MetaboAnalyst 5.0数据库分析可能的代谢途径,从代谢水平分析MQ治疗AB的作用机制。结果显示MQ能明显改善模型大鼠支气管和肺组织的病理损伤,降低NLRP3和p38 MAPK的mRNA水平(P < .05, .01),降低血清和肺匀浆中TNF-α、IL-6和IL-1β的水平(P < .05,.01)水平,提高血清和肺匀浆中的 IL-10 (P < .0001,.01)水平,并通过甘油磷脂代谢途径调节 AB 大鼠体内的异常代谢物。结论MQ能明显改善混合烟雾联合肺炎链球菌诱导的AB,其诱导机制可能与甘油磷脂代谢失调有关,该研究为MQ在临床上的合理应用提供了参考。
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Maqin Xiaoke Granule Alleviates Acute Bronchitis Induced by Smoking Combined With Streptococcus pneumoniae by Regulating NLRP3/p38 MAPK/Glycerophospholipid Pathway
Objective: To explore the efficacy and mechanism of action of Maqin Xiaoke Granules (MQs) in the treatment of acute bronchitis (AB). Methods: The rat model of AB was established by mixed smoke and Streptococcus pneumoniae nasal infection. The pathological changes of lung tissue and bronchus were observed by hematoxylin-eosin (HE) staining and scored. The mRNA expression levels of NOD-like receptor thermal protein domain associated protein 3 ( NLRP3) and p38 mitogen-activated protein kinase ( p38 MAPK) in lung tissue were detected by real-time fluorescence quantitative polymerase chain reaction, and the expression levels of inflammatory factors (tumor necrosis factor-α [ TNF-α], interleukin-6 [ IL-6], interleukin-1β [ IL-1β], interleukin-10 [ IL-10]) in serum and lung homogenate were detected by enzyme-linked immunosorbent assay. The therapeutic effect of MQ on rats with AB was preliminarily evaluated. Metabolic profiling of rat serum was performed by ultra high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry/mass spectrometry, combining P < .05, variable important in projection > 1, fold change > 1.5 and screening for differential metabolites based on univariate and multivariate statistical analyses, and then analyzing the possible metabolic pathways with the help of MetaboAnalyst 5.0 database, to analyze the mechanism of MQ’s action on the treatment of AB at the metabolic level. Results: MQ significantly ameliorated the pathological injury of bronchial and lung tissues in model rats, reduced the mRNA levels of NLRP3 and p38 MAPK ( P < .05, .01), lowered the levels of TNF-α, IL-6, and IL-1β in serum and lung homogenates ( P < .05, .01), and elevated IL-10 in serum and lung homogenates ( P < .0001, .01) levels, and to regulate abnormal metabolites in rats with AB via the glycerophospholipid metabolic pathway. Conclusion: MQ significantly improved AB induced by mixed smoke combined with Streptococcus pneumoniae, and the mechanism of its induction may be related to the dysregulation of glycerophospholipid metabolism, and this study provides a reference for the rational clinical application of MQ.
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