从生物力学研究温化寒痰法抑制肿瘤转移的机制

The idea of “Phlegm is flowing away in nature”in traditional Chinese medicine is similar to the conception of tumor metastasis in modern medicine. “If disease resulted from Phlegm-retention, warm Yang drug can treat it.” Therefore, we hypothesize that warm and relieve Cold-Phlegm therapy is an important principle of treating tumor metastasis. In the opinion of certain scholars, warm Yang drug could stimulate the growth and metastasis of tumor and shouldn’t be used to treat tumor. However, our previous reports indicated that the combination of warm Yang drug Radix Aconiti Praeparata and eliminate Phlegm drug Rhizoma Bolbostemmatis, with optimum drug dosage and weight ratio, could enhance the inhibitory effect of Rhizoma Bolbostemmatis on the migration of breast cancer, as well as antagonize the promoting effect of Radix Aconiti Praeparata on it. Our results suggested that the key of applying warm Yang therapy to treat breast cancer is combining warm Yang drug and eliminate Phlegm drug appropriately. In the present study, median effect principle is used to evaluate the nature of the interaction between Radix Aconiti Praeparata and Rhizoma Pinelliae/Fructus Trichosanthis/Rhizoma Bolbostemmatis and select the conditions-directed synergism in inhibiting tumor cells migration, including drug dosage and compatibility proportion. From the field of biomechanics, the effect of Radix Aconiti Praeparata in combination with Rhizoma Pinelliae/ Fructus Trichosanthis/ Rhizoma Bolbostemmatis on tumor mechanical properties, including cell adhesion, deformability, migration, invasion and cytoskeleton are observed on the molecular, cellular and overall levels. This study contributes to reveal the mechanism of warm and relieve Cold-Phlegm therapy inhibiting tumor metastasis and provide the basis for apply this therapeutic method to treat tumor clinically.

中医认为“痰性流走”，与现代医学对恶性肿瘤易于转移的认识相似，而“病痰饮者，当以温药和之”，故温化寒痰法是防治肿瘤转移的重要治法。有学者认为温阳药物可以刺激肿瘤生长转移，在肿瘤治疗中不宜使用。但是我们发现，温阳药物附子与化痰药物土贝母在适宜条件下配伍，可增强土贝母抑制乳腺癌细胞迁移的作用，同时拮抗附子促进细胞迁移的不良作用，提示运用温阳法治疗乳腺癌的关键在于如何将温阳药物与化痰药物合理配伍。本研究运用中效原理定量分析温阳药物附子配伍化痰药物半夏/瓜蒌/土贝母对肿瘤细胞迁移的影响，筛选产生最大协同抑制效应的药物配伍条件，包括药物剂量与配伍比例；从生物力学角度，观察附子配伍半夏/瓜蒌/土贝母在分子、细胞和整体水平，对肿瘤粘附、变形、迁移、侵袭、细胞骨架等力学特性的影响。研究结果有助于揭示温化寒痰法抑制肿瘤转移的机制，并为临床正确运用温阳法防治肿瘤转移提供依据。

中医认为“痰性流走”，与现代医学对恶性肿瘤易于转移的认识相似，而“病痰饮者，当以温药和之”，故温化寒痰法是防治肿瘤转移的重要治法。本研究立足于传统中医痰病学说，结合现代科学技术，选择具有温阳作用的代表性药物附子、砒霜和具有化痰作用的代表性药物土贝母、栝蒌，运用中效原理定量分析附子与土贝母水提物、栝蒌中主要化学成分葫芦素B（CuB）分别与砒霜的主要化学成分三氧化二砷（ATO）/附子的主要成分去甲乌药碱，在不同剂量与比例下配伍使用的抗肿瘤作用，筛选产生协同作用的药物配伍条件并分析其作用机制。研究发现，附子与土贝母的水提物合理配伍可以通过下调黏附分子E-钙粘蛋白表达、改变骨架蛋白actin微丝重组与分布，协同抑制乳腺癌细胞迁移；栝蒌中的CuB通过抑制STAT3和Erk磷酸化，上调肿瘤细胞凋亡相关蛋白Bax/Bcl -2的比例，增强ATO促进淋巴瘤凋亡的作用，在裸鼠移植性淋巴瘤模型中，CuB与ATO在一定条件下配伍使用时，抑瘤率也高于两种药物单用；CuB可抑制乳腺癌细胞生长，诱导细胞凋亡，阻滞细胞周期于G2/M 期。而与去甲乌药碱合理配伍时在体内外可以增强Cu B的抗乳腺癌作用。通过网络药理学的研究方法，我们预测出CuB和去甲乌药碱作用的疾病靶点蛋白，并通过western blot的方法验证，发现CuB和去甲乌药碱通过抑制AKT与CDK2相互作用来协同抑制乳腺癌细胞增殖；我们还从力学的角度探究了化痰药物栝蒌中的CuB的抑制乳腺癌迁移的效果。研究发现CuB通过改变骨架蛋白在细胞内的分布和重组，改变细胞粘附性和变形性的力学特性，进而抑制乳腺癌细胞的迁移和侵袭。其机制是CuB抑制整合素下游Rho家族GTPases RAC 1/Cdc 42/RhoA的信号转导，下调乳腺癌细胞F-actin/vimentin/FAK/vinculin的表达。可见，运用温阳药物治疗肿瘤的关键在于将温阳药物与化痰药物合理配伍，从而实现“病痰饮者，当以温药和之”的治疗法则；且温化寒痰法可通过改善肿瘤生物力学性质抑制肿瘤转移，为临床正确运用温化寒痰法防治肿瘤提供依据。