目的 基于转录组学分析小青龙汤治疗慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）的作用机制。方法 雄性C57BL/6小鼠随机分为对照组、模型组、地塞米松（1 mg/kg）组和小青龙汤低、中、高剂量（1.46、0.73、0.36 g/kg）组，每组8只。除对照组外，其余各组小鼠采用烟草提取物联合脂多糖鼻内滴注法构建COPD模型。连续给药4周，观察各组小鼠的一般状态，采用苏木素-伊红（HE）染色观察小鼠肺组织病理变化，采用ELISA测定血清中炎性因子水平；采用转录组学测序技术分析对照组、模型组和小青龙汤高剂量组小鼠肺组织的基因表达谱，并对显著差异表达的基因进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析；采用qRT-PCR和Western blotting检测小鼠肺组织中环磷腺苷（cyclic adenosine monophosphate，cAMP）信号通路相关基因及蛋白的表达。结果 与对照组比较，模型组小鼠一般状态差，肺组织可见明显病理损伤，血清中炎性因子水平显著升高（P＜0.01）；与模型组比较，各给药组小鼠一般状态、肺组织损伤情况均得到不同程度的改善，血清中炎性因子水平显著降低（P＜0.05、0.01），其中小青龙汤高剂量组和地塞米松组改善作用最为明显。转录组学分析结果表明，对照组vs模型组和模型组vs小青龙汤高剂量组差异表达基因广泛参与了cAMP信号通路、神经活性配体-受体相互作用、昼夜节律等相关信号通路。qRT-PCR和Western blotting结果显示，与对照组比较，模型组小鼠肺组织中cAMP、PKA、CREB表达显著降低（P＜0.01），VEGF表达显著升高（P＜0.01）；与模型组比较，小青龙汤高剂量组小鼠肺组织中cAMP、PKA、CREB表达显著升高（P＜0.05、0.01），VEGF表达显著降低（P＜0.01）。结论 小青龙汤可显著减少COPD小鼠气道炎性细胞浸润、减轻肺部气道重塑，并通过调控cAMP信号通路，治疗COPD。

Objective To investigate the mechanism of Xiaoqinglong Decoction (小青龙汤) in treating chronic obstructive pulmonary disease (COPD) based on transcriptomics analysis. Methods Male C57BL/6 mice were randomly divided into control group, model group, dexamethasone (1 mg/kg) group, and Xiaoqinglong Decoction low-, medium-, high-dose (1.46, 0.73, 0.36 g/kg) groups, with eight mice in each group. Except for the control group, all other groups of mice were used to construct COPD models by intranasal infusion of tobacco extract combined with lipopolysaccharide. After continuous administration for four weeks, the general status of mice in each group were observed, hematoxylin-eosin (HE) staining was used to observe the pathological changes in lung tissue of mice, ELISA was used to measure the levels of inflammatory factors in serum; Transcriptome sequencing technology was used to analyze the gene expression profiles of lung tissues in control group, model group and Xiaoqinglong Decoction high-dose group, gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed on significantly differentially expressed genes; qRT-PCR and Western blotting were used to detect the expressions of genes and proteins related to cyclic adenosine monophosphate (cAMP) signaling pathway in lung tissue. Results Compared with control group, mice in model group showed poor general condition, significant pathological damage in lung tissue, and significantly increased levels of inflammatory factors in serum (P < 0.01); Compared with model group, the general condition and lung tissue damage of mice in each treatment group were improved to varying degrees, and the levels of inflammatory factors in serum were significantly reduced (P < 0.05, 0.01). Among them, Xiaoqinglong Decoction high-dose group and dexamethasone group had the most significant improvement effect. The transcriptome analysis results showed that the differentially expressed genes between control group and model group, as well as Xiaoqinglong Decoction high-dose group were widely involved in cAMP signaling pathways, neuroactive ligand-receptor interactions, circadian rhythms and other related signaling pathways. qRT-PCR and Western blotting results showed that compared with control group, the expressions of cAMP, PKA and CREB in lung tissue of mice in model group mice were significantly reduced (P < 0.01), while the expression of VEGF was significantly increased (P < 0.01); Compared with model group, Xiaoqinglong Decoction high-dose group significantly increased the expressions of cAMP, PKA and CREB in lung tissue of mice (P < 0.05, 0.01), and significantly decreased the expression of VEGF (P < 0.01). Conclusion Xiaoqinglong Decoction could significantly reduce inflammatory cell infiltration in the airways of COPD mice, alleviate lung airway remodeling, and treat COPD by regulating cAMP signaling pathway.

R285.5

甘肃省高等学校产业支撑计划项目（2022CYZC-53）;甘肃省青年博士支持项目（2023QB-092）;兰州市科技计划项目（2023-ZD-151）