Cang Song, Liu Ran, Jin Wei, Tang Qi, Li Wanjun, Mu Kunqian, Jin Pengfei, Bi Kaishun, Li Qing
School of Pharmacy, National and Local Joint Engineering Laboratory for Key Technology of Chinese Material Medica Quality Control, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110016, China.
School of Applied Chemistry and Biological Technology, Shenzhen Polytechnic, 7098 Lau sin Avenue, Shenzhen, 518000, China.
Chin Med. 2021 Nov 27;16(1):126. doi: 10.1186/s13020-021-00535-x.
Lung cancer remains the leading cause of mortality from malignant tumors, non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer cases, and individualized diagnosis and treatment is an effective trend. The individual characteristics of different traditional Chinese medicine (TCM) syndromes of NSCLC patients may be revealed by highly specific molecular profiles.
In this study, 10 NSCLC patients with Qi deficiency and Yin deficiency (QDYD) syndrome and 10 patients with Qi deficiency of lung-spleen (QDLS) syndrome in TNM stage III-IV as well as 10 healthy volunteers were enrolled. Aiming at the varied syndromes of NSCLC patients with "Yin deficiency" as the main difference, a proteomics research based on data-independent acquisition (DIA) was developed. Of the dysregulated proteins in NSCLC patients, lipid metabolism was significantly enriched. Thereafter, nontargeted lipidomics research based on UPLC-Q-TOF/MS was performed in 16 patients, with 8 individuals randomly selected from each syndrome group. Furthermore, the considerably different characteristics between the syndromes and pathological mechanisms of NSCLC were screened by statistical and biological integrations of proteomics and lipidomics and the differential metabolic pathways of the two similar syndromes were further explored. Besides, lipids biomarkers were verified by a clinically used anticancer Chinese medicine, and the level of key differential proteins in the two syndromes was also validated using ELISA.
The results showed that glycerophospholipid metabolism, sphingolipid metabolism, glycolipid metabolism, and primary bile acid biosynthesis were altered in NSCLC patients and that glycerophospholipid metabolism was significantly changed between the two syndromes in lipidomics analysis. Among the proteins and lipids, ALDOC and lysophosphatidylcholine (LPCs) were revealed to have a strong relationship by statistical and biological integration analysis, and could effectively distinguish QDLS and QDYD syndromes. Notably, the patients with different syndromes had the most typical metabolic patterns in glycerophospholipid metabolism and glycolysis, reflecting the differences in the syndromes dominated by "Yin deficiency".
ALDOC and LPCs could be employed for the differentiation of NSCLC patients with QDLS and QDYD syndromes, and "Yin deficiency" might be associated with glycerophospholipid metabolism and glycolysis pathway. The results provided a theoretical basis for "Syndrome differentiation" in TCM diagnosis. Moreover, the developed integrated strategy could also provide a reference for individualized diagnosis and treatment of other diseases.
肺癌仍然是恶性肿瘤致死的主要原因，非小细胞肺癌（NSCLC）占肺癌病例的大多数，个体化诊断和治疗是一种有效的趋势。NSCLC患者不同中医证候的个体特征可能通过高度特异性的分子图谱得以揭示。
本研究纳入10例TNM分期为III-IV期的气虚阴虚（QDYD）证NSCLC患者、10例肺脾气虚（QDLS）证NSCLC患者以及10名健康志愿者。针对以“阴虚”为主要差异的NSCLC患者的不同证候，开展了基于数据非依赖采集（DIA）的蛋白质组学研究。在NSCLC患者中失调的蛋白质中，脂质代谢显著富集。此后，对16例患者进行了基于超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF/MS）的非靶向脂质组学研究，每个证候组随机选取8例个体。此外，通过蛋白质组学和脂质组学的统计与生物学整合，筛选出NSCLC证候与病理机制之间的显著差异特征，并进一步探索两种相似证候的差异代谢途径。此外，用一种临床使用的抗癌中药验证脂质生物标志物，并用酶联免疫吸附测定（ELISA）法验证两种证候中关键差异蛋白的水平。
结果显示，NSCLC患者的甘油磷脂代谢、鞘脂代谢、糖脂代谢和初级胆汁酸生物合成发生改变，脂质组学分析表明两种证候之间甘油磷脂代谢有显著变化。通过统计和生物学整合分析发现，蛋白质和脂质中的醛缩酶C（ALDOC）和溶血磷脂酰胆碱（LPCs）有很强的相关性，并且能够有效区分QDLS和QDYD证候。值得注意的是，不同证候的患者在甘油磷脂代谢和糖酵解方面具有最典型的代谢模式，反映了以“阴虚”为主的证候差异。
ALDOC和LPCs可用于鉴别QDLS和QDYD证的NSCLC患者，“阴虚”可能与甘油磷脂代谢和糖酵解途径有关。研究结果为中医诊断中的“辨证”提供了理论依据。此外，所建立的整合策略也可为其他疾病的个体化诊断和治疗提供参考。
