目的 基于灵芝改善急性肺损伤（acute lung injury，ALI）的药效作用，建立不同产地灵芝指纹图谱，通过“物质-功效”相关性分析揭示灵芝质量标志物（quality marker，Q-Marker）。方法 采用超高效合相色谱-四极杆飞行时间串联质谱（ultra-performance convergence chromatography coupled to quadrupole time-of-flight tandem mass spectrometry，UPC²-Q-TOF-MS/MS）技术，在正、负离子模式下建立15批不同产地灵芝的指纹图谱，并对共有峰进行指认。建立脂多糖诱导的ALI小鼠模型，给药干预后，以血清和支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-1β、髓过氧化物酶（myeloperoxidase，MPO）含量及肺组织湿干质量比为药效指标，综合灰色关联度分析、双变量相关分析、正交偏最小二乘分析3种方法进行“物质-功效”相关性分析，筛选出灵芝干预ALI的潜在Q-Marker，并测定Q-Marker含量。结果 15批灵芝指纹图谱在正离子模式下相似度为0.678～0.835，共有峰18个；在负离子模式下相似度为0.645～0.899，共有峰22个。药效实验表明15批不同产地灵芝对ALI均有一定保护作用。筛选出灵芝酸F、灵芝酸DM、灵芝烯酸D、灵芝酸A、灵芝酸C2、棕榈酸乙酯共6个潜在Q-Marker。选取药效最优的S12（产地为黑龙江牡丹江市）灵芝开展潜在Q-Marker含量测定，其中灵芝酸A含量最高，为（1.328 2±0.025 1）mg/g；灵芝烯酸D含量最低，为（0.186 3±0.003 5）mg/g。结论 通过“物质-功效”相关性分析筛选出与灵芝干预ALI药效具有显著关联性的6个潜在Q-Marker，为灵芝的质量评价、品质控制及药效物质基础研究提供了科学依据，也为灵芝在ALI防治中的进一步开发应用奠定了基础。

Objective To establish finger print of Ganoderma lucidum from different origins and identify quality marker (Q-Marker) through “compound-effect” correlation analysis based on the pharmacological effect of Ganoderma lucidum in improving acute lung injury (ALI). Methods Ultra-performance convergence chromatography coupled to quadrupole time-of-flight tandem mass spectrometry (UPC²-Q-TOF-MS/MS) technology was used to establish fingerprint spectra of 15 batches of G. lucidum from different origins in positive and negative ion modes, and common peaks were identified. A mouse model of lipopolysaccharide-induced ALI was established, after drug intervention, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, myeloperoxidase (MPO) in serum and bronchoalveolar lavage fluid (BALF) and lung tissue wet dry mass ratio were used as pharmacological indicators. Comprehensive grey correlation analysis, bivariate correlation analysis and orthogonal partial least squares analysis were used to conduct “substance-efficacy” correlation analysis, potential Q-Markers of G. lucidum intervention in ALI were screened, and the contents of Q-Marker were measured. Results The similarity of 15 batches of G. lucidum fingerprint spectra in positive ion mode ranges from 0.678 to 0.835, with a total of 18 peaks. In negative ion mode, the similarity ranges from 0.645 to 0.899, with a total of 22 peaks. Pharmacological experiments showed that 15 batches of G. lucidum from different origins had a certain protective effect on ALI. A total of six potential Q-Markers were identified, including ganoderic acid F, ganoderic acid DM, ganoderic acid D, ganoderic acid A, ganoderic acid C2 and ethyl palmitate. The most effective G. lucidum with S12 (produced in Mudanjiang city, Heilongjiang Province) was selected for potential Q-Marker content determination, the content of ganoderic acid A was the highest, at (1.328 2 ± 0.025 1) mg/g, the content of ganoderic acid D was the lowest, at (0.186 3 ± 0.003 5) mg/g. Conclusion Through “substance-effect” correlation analysis, six potential Q-Markers were identified as significantly associated with the pharmacological effects of G. lucidum in the treatment of ALI. This provides a scientific basis for the quality evaluation, quality control, and research on material basis of the pharmacologically active components of G. lucidum, and lays the foundation for its further development and application in the prevention and treatment of ALI.

R282.6

黑龙江省重点研发计划项目（2024ZX10B05）；中央支持地方高校改革发展资金人才培养项目；浙江省基础公益研究计划（LTGN23H280003）；宁波市社会公益研究第二批项目（2023S058）