西黄丸通过影响甘氨酸代谢抑制乳腺癌小鼠肿瘤生长的机制研究

Mechanism of Xihuang Wan(西黄丸) Inhibiting Tumor Growth in Breast Cancer Mice by Affecting Glycine Metabolism

目的:研究西黄丸对荷4T1乳腺癌小鼠甘氨酸代谢的影响,观察肿瘤生长情况,进而探讨药物抑瘤作用新途径。方法:建立4T1小鼠乳腺癌荷瘤模型,造模第2天西黄丸连续灌胃给药14 d,第15天眼球取血,剥离荷瘤组织进行称质量、切片、匀浆。采用HPLC-MS检测血浆中甘氨酸浓度;称质量法计算抑瘤率;TUNEL染色测肿瘤细胞凋亡;体外实验测甘氨酸对肿瘤细胞增殖、凋亡的影响。结果:模型组血浆甘氨酸浓度明显低于正常组(P<0.05),西黄丸组血浆甘氨酸浓度明显高于模型组(P<0.01);西黄丸组抑瘤率为33.3%;TUNEL染色显示西黄丸组肿瘤细胞凋亡增加;含甘氨酸培养基较正常培养基肿瘤细胞增殖明显;早、晚期凋亡均减少。结论:西黄丸可通过抑制肿瘤细胞对甘氨酸的摄取,抑制细胞增殖、促进细胞凋亡,进而抑制荷瘤生长。

Objective： To study the effect of Xihuang wan on glycine metabolism in 4 T1 breast cancer mice, observe the tumor growth, and then explore the new ways of drug antitumor action. Methods： A 4 T1 tumor-bearing model was established. And the Xihuang wan gavage was given in the second day and last for 14 days, and the blood samples were taken from the eyeballs at the 15 thday. The tumor tissue was peeled off then weighed, sliced and homogenized. The concentration of glycine in plasma was measured by LC-MS. The tumor growth rate was measured by TUNEL staining. Effect of glycine on the apoptosis of tumor cells in vitro. Results： The model group showed lower plasma glycine concentration than normal group（P〈0.05）; while the Xihuang wan group sowed higher ratio of glycine than the model group（P〈0.01）; The tumor suppressor rate was33.3% in the Xihuang wan group. TUNEL staining showed that the apoptosis of tumor cells in Xihuang wan group increased; The glycine-containing medium had more living cells with less early or late apoptosis in glycine than glycine-free medium. Conclusion： Xihuang wan can decrease the glycine metabolism and increase the apoptosis of tumor cells thus inhibiting tumor growth.