The Mechanism Research Of Xihuang Pills Inhibiting Tumor Growth In Breast Tumor-bearing Mice By Regulating Amino Acid Metabolism

Objective: To study the effect of Xihuang Pills on amino acid metabolism in 4T1 breast tumor-bearing mice,observe the tumor growth,and then explore the new ways of drug antitumor action.Methods: Thirty female BABL/c mice are randomly divided into three groups,the author names them “the normal group”,“the model group”,and “the Xihuang Pills group”.4T1 tumor-bearing models are established among the group of model and Xihuang Pills.The XiHuang Pills gavage is given in the second day obeying to the recipe that twice a day and 0.2ml each time which lasts for 14 days,while the model are given distilled water in the same volume.All mice are given sufficient food and water.The blood samples are taken from the eyeballs at the 15 th day,the mice are put to death by cervical dislocation.The tumor tissue is peeled off then weighed,sliced and homogenized.The author measures the concentration of glycine in plasma by LC-MS,while calculates the inhibitory rate by mass method,The effects of glycine on proliferation and apoptosis of tumor cells are tested in vitro.At the same time,the model identification method of partial least squares-discriminant analysis(PLD-DA)is used to study the concentration of amino acids in the blood of mice,The author establishes the amino acid multidimensional statistical screening model to screen the relevant differential amino acids of plasma of the tumor-bearing mice.The author uses the CCK8 kit to test the effect of this amino acid which is screened in last step on the proliferation and apoptosis of 4T1 breast cancer cells.The LDH kit and ATP kit are used to find out the effect on the expression of LDH of the amino acid above.Results: Compared with the normal group,the plasma glycine concentration in the model group is lower(P <0.05)while the ratio of glycine in the Xihuang Pills group is higher than that in the model group(P <0.01);TUNEL staining showes that the apoptosis of tumor cells in Xihuang Pills group increases;The glycine-containing medium has more living cells with less early nor late apoptosis than glycine-free medium.β-alanine is the amino acid that screened by the PLD-DA,and in vitro experiment β-alanine-containing medium has fewer living cells with more cell apoptosis thanβ-alanine-free medium.Conclusions: Xihuang Pills can increase the apoptosis of tumor cells thus inhibiting tumor growth.Which is due to its effect to decrease the glycine metabolism,while increase the β-alanine level.