白花蛇舌草-半枝莲药对通过有氧糖酵解途径协同抗肿瘤的机理研究

Hedyotis diffusa Willd.-Scutellaria barbata D. Don herbal pair was clinically proved to be effective against many tumors in traditional Chinese medicine. Although studies on Hedyotis diffusa Willd. or Scutellaria barbata D. Don single herb have been further developed, there are few studies on the compatibility of the herbal pair. Our previous studies showed that Hedyotis diffusa Willd. and Scutellaria barbata D. Don effective parts exhibit synergistic antitumor activity, which might be related with aerobic glycolysis, and further studies are needed. This project intent to study the relationship between the antitumor activities and fingerprints of different Hedyotis diffusa Willd.-Scutellaria barbata D. Don compatibility using modern separation analysis techniques and drug screening models. On that basis, we will screen the optimal antitumor effective parts of Hedyotis diffusa Willd. and Scutellaria barbata D. Don herbal pair, analyze their chemical components, and investigate their effects on tumor aerobic glycolysis, and then further explore their effects on key enzymes of aerobic glycolysis, mitochondria function, and expressions of oncogenes and tumor supressors, as well as signal pathways such as PI3K/AKT/mTOR, AMPK, to illustrate the exact mechanism and targets of the herbal pair compatibility on aerobic glycolysis, providing theoretical reference for clinical usages. This project will not only lay basis for the theory development from herb compatibility to effective parts compatibility of herbal pairs and complex prescriptions, but also provide important references for the modern research of new complex prescriptions and preparations of traditional Chinese medicine.

白花蛇舌草-半枝莲药对经长期临床验证对多种肿瘤疗效确切。目前白花蛇舌草、半枝莲单味药的研究较广泛，但白花蛇舌草-半枝莲药对配伍抗肿瘤的研究甚少。我们前期研究显示白花蛇舌草、半枝莲有效部位可协同抑制肿瘤，其机制与有氧糖酵解有关，有待于进一步研究。故本课题拟结合现代分离分析技术和活性筛选模型，研究白花蛇舌草、半枝莲不同配比的抗肿瘤活性与指纹图谱间的关系，确定最佳配比。基于此，筛选出药对协同抗肿瘤的最优部位及组合，进行成分分析并探讨其对肿瘤有氧糖酵解的影响，进而分析其对糖酵解关键酶、线粒体功能、相关癌基因/抑癌基因及PI3K/AKT/mTOR、AMPK等信号通路的影响，以期阐明白花蛇舌草-半枝莲药对配伍影响肿瘤有氧糖酵解的确切机制及靶点，为临床提供理论参考。这将为中药药对或复方从药材配伍到有效部位配伍的理论发展奠定基础，同时也可为有关中药新复方、新制剂的现代化研究提供重要参考。

白花蛇舌草-半枝莲药对经长期临床验证对多种肿瘤疗效确切。目前白花蛇舌草、半枝莲单味药的研究较广泛，但白花蛇舌草-半枝莲药对配伍抗肿瘤的研究甚少。本项目主要进行了以下研究：1）制备了白花蛇舌草、半枝莲单药及不同配比的水煎液浸膏；体内活性筛选结果表明二者1:1配伍的浸膏具有更好体内外抗肿瘤效果，且对小鼠的体重影响较小。初步的机制研究发现二者配伍的抗肿瘤作用与糖酵解有关。2）进一步制备了白花蛇舌草有效部位（HD）、半枝莲有效部位（SB）、白花蛇舌草-半枝莲 1:1 配伍有效部位（HD-SB），测定了其总黄酮含量，超高压液相色谱/质谱(Q-TOF/MS)鉴定了其主要成分。3）活性评价结果发现，与SB、HD相比，相同剂量下HD-SB具有更显著抗肿瘤作用，且HD-SB对小鼠体重及重要脏器均无明显影响；3）SB与HD单用及联合抗肿瘤作用评价发现，1:1的SB与HD配伍体外协同抗肿瘤作用最明显；二者联用的体内抑瘤作用显著优于单药；与对照组比较，两药单用、联用均可显著抑制葡萄糖摄取、乳酸分泌并降低PKM2/PKM1比率，且联用效果更显著；4）网络药理学分析结果表明，槲皮素、木犀草素、汉黄芩素和谷固醇可能是其抗肿瘤的主要有效成分，其抗肿瘤的主要靶点有：PTGS2、HSP90、EGFR、MMP2、PPAR-γ、GSTP、CDK2、PIK3K等。进一步的验证结果表明，HD-SB抗肿瘤的主要机制与EGFR、PPARγ、PI3K/AKT等介导的凋亡信号通路有关；5）我们还对其重塑肿瘤微环境的靶点和机制进行了更深入的网络药理学挖掘，预测结果后续将进行进一步的实验验证。基于该药对的临床应用、药性特点、网络药理学及我们初步的研究结果，提出了新的假说：该药对可能是通过重塑肿瘤微环境来发挥抗肿瘤作用的。以上结果为该药对从药材配伍到有效部位配伍的发展奠定了基础，也可为其进一步开发了利用提供了重要参考。