从干预有氧糖酵解的角度探讨丹参通过活血化瘀治疗肺癌的药理机制

Glycolysis is the main pathway of tumor cells to produce energy even oxygen exists, which called aerobic glycolysis. Glycolysis could survive cancer cells from hypoxia during tumorigenesis and induce chemoresistance, which makes it became a new target for cancer therapy. “blood stasis” exists in most cancer patients because “Qi is deficiency and unification of the blood”. “Qi” refers the nutrients and the energy. Therefore, aerobic glycolysis resulting energy deficiency supposed to be one of the causes to induce “blood stasis” in cancer patients. Besides, as a classic “huo xue hua yu” herbs, Danshen could reverse the glycolysis which makes lung cancer cells cannot adapt to the hypoxic environment and die. This is the hypothesis in the study. To confirm it, cell lines and model animals are operated to figure out the effects of Danshen on the glycolysis in lung cancer cell and the role of aerobic glycolysis in Danshen’s pharmacologic function during lung cancer treating. Metabolomics test, bioinformatic analysis, and molecular biologic array are used to study the mechanism of Danshen regulates the aerobic glycolysis in lung cancer cell. In this project, the purpose of the study on the Danshen’s pharmacologic mechanism is to discover the intrinsic relation between traditional medicine theory and modern scientific connotation. It is an innovative attempt to integrate traditional Chinese medicine and Western medicine.

无论缺氧与否，糖酵解均为肿瘤细胞优先选择的能量代谢途径，即有氧糖酵解，其在肿瘤的发生发展和诱导药物耐受中具有重要作用，现已成为癌症治疗的新靶点。中医认为“气”与能量密切相关，癌症多为“脏腑亏损、气虚统摄无权而生血瘀”。研究表明血瘀证型的癌症患者葡萄糖代谢率更高。我们推测肿瘤细胞低效率产能的有氧糖酵解代谢方式与中医的“气虚血瘀”之间必然存在联系；活血化瘀代表中药丹参可通过调控有氧糖酵解水平使肺癌细胞无法适应乏氧环境而死亡以治疗肺癌。本课题拟通过系统的细胞及动物实验分析丹参对肺癌细胞有氧糖酵解水平的影响；研究有氧糖酵解在丹参抗肺癌中的作用；并联合代谢组学技术、生物信息学手段和分子生物学方法阐明丹参干预有氧糖酵解的分子机制。本项目旨在通过深度解读丹参“活血化瘀”防治肺癌的现代科学内涵，探索中医理论和西医科学之间的内在联系，是中西医结合的一次创新性尝试。

肿瘤细胞发生代谢重编程。即使氧气充足，肿瘤细胞依然选择糖酵解为主要的能量代谢途径，即有氧糖酵解。有氧糖酵解在肿瘤发生发展中发挥重要作用，现已成为癌症治疗的新靶点。中医临床发现癌症患者多为“脏腑亏损、气虚血瘀”之证。通过前期研究我们认为肿瘤细胞低效率产能的有氧糖酵解代谢与中医的“气虚血瘀”之间必然存在联系；活血化瘀代表中药丹参可通过调控有氧糖酵解水平使肺癌细胞无法适应乏氧环境而死亡而治疗肺癌。通过课题的研究，已明确丹参可通过调控肺癌的有氧糖酵解水平发挥抗肺癌作用。体内实验结果显示，丹参水提液、丹参醇提液及丹参活性成分丹参素、二氢丹参酮I、丹参酮IIA均下调HKII、PKM等糖酵解关键酶的表达，抑制肺癌细胞的有氧糖酵解水平，发挥抗肺癌作用。在细胞实验中，丹参素和二氢丹参酮I明显抑制HKII、PKM、PFK的表达，上调PDH表达。使用HKII抑制剂进行预处理后，发现丹参素和二氢丹参酮I可以增强抑制剂对HKII的抑制作用，有氧糖酵解水平继续下调，肿瘤细胞增殖减少，凋亡明显增多。通过siRNA和shRNA分别下调和上调HKII表达后，发现丹参素和二氢丹参酮I可继续下调HKII表达水平，但在HKII高表达细胞株中，有氧糖酵解水平得以保持，细胞增殖能力恢复，凋亡减少；但在HKII低表达细胞的结果与化学抑制剂的作用一致。通过进行化学干预和转基因技术干预实验证实，丹参的两种活性成分抑制肺癌的有氧糖酵解水平和抗肺癌作用与其抑制HKII的作用密切相关。本研究从肿瘤特殊代谢方式——有氧糖酵解的角度解析了丹参治疗肺癌的药理作用，研究了中医“气虚血瘀”的科学内涵。本项目通过研究丹参对肺癌细胞有氧糖酵解的作用及分子机制，探索中医临床通过研究“活血化瘀”中药丹参的药理机制，进一步阐述了丹参及活血化瘀治法治疗肺癌的内在机制，为开发有效药物，丰富中药治疗的疗效机理，和肺癌的防治探索一条新的思路和途径。相关的研究结果已发表高水平论文4篇，其中3篇为SCI收录，另有一篇文章正在投稿中。整个研究过程有两名硕士研究生完成，一名已顺利毕业，另一名同学已进入研究生毕业论文撰写阶段。