Study On Anti-tumor Activity Of Effective Components In Cantharis Metabolic Liquid

Blister beetles is a dry body of Meloidae insects like Mylabris phalerata Pallas or Mylabris Cichorii Linnaeus.Modern pharmacological studies showed that cantharis has good anti-tumor effect,but also has certain toxicity of kidney and liver.Our research group has demonstrated for the first time that PSB bioconversion of the blister beetle is able to reduce the toxicity of a vital Chinese medicine commonly used in anticancer treatments,and can improve it's antitumor active in vitro.In order to explore the material basis of antitumor activity of cantharis metabolic liquid,we activity-guided to extract and separate of metabolic liquid,obtained a series of monomers,but they are not strong antitumor activity in vitro.Considering to be a variety of active ingredients for the synergistic anti-tumor effect,then start from the concept of traditional Chinese medicine,combined with the research ideas of group of Chinese medicine,select three activity compounds which antitumor activity relatively strong in cantharis metabolic liquid.Effective components as a whole to study its antitumor activity,through the antitumor activity of the effective components to understand why the cantharis metabolic liquid antitumor activity is enhanced.1.The research group of the organic synthesis of the three compounds were compared with the laboratory's to determine whether the synthetic material is the three activity compounds which antitumor activity relatively strong in cantharis metabolic liquid.Then we through the HPLC method to study the proportion of three compounds in cantharis metabolic liquid.The results showed that the ratio of 5:1:2,then the ratio was uesed in the following experiments.2.In order to explore whether the effective compounds antitumor activity is enhanced in vitro and to which cell has the stronger effect.On the basis of previous research,we chose lung cancer A549,human HepG-2,breast cancer MCF-7 three tumor cells.The results showed that the ratio of drug antitumor activity significantly enhanced in vitro and the inhibitory effect of effective compounds to lung cancer A549 is the best.So in the follow-up study,we chose the lung cancer model to carry on the anti-tumor experiment in vivo.3.Induced apoptosis is an important means for the treatment of tumors,in order to explore whether effective compounds in vitro antitumor activity is enhanced by inducing apoptosis to achieve,the experimental cell apoptosis by morphological observation,AnnexinV/PI method was used to detect the content of DNA double staining method to detect cell apoptosis,and apoptosis was determined during the period.Cell morphology and AnnexinV/PI double staining results showed that effective compounds can induce lung cancer cell apoptosis;the detection result of DNA content showed that the effective compounds in vitro causes apoptosis is blocked the cell cycle in G2 phase,thus preventing cell proliferation.4.The effective compounds by inducing apoptosis has a strong inhibitory effect on lung cancer cells in vitro.In order to further study the effective compounds safety and for the subsequent in vivo antitumor experimental dosage provides reliable experimental basis,the ratio of drug on acute toxicity of mice with lethal dose test method for the determination of the acute toxicity and LD50.The results showed that the effective compounds LD50=214mg/kg.Our previous study found that the water extract of beetle LD50= 206mg/kg,and the main effective component of cantharidin LD50= 1.86mg/kg.The toxicity of the effective compounds decreased 115 times compared with cantharidin,and the toxicity of the effective compounds is lower than beetle water extraction liquid.5.To establish the Lewis mouse lung cancer ascites tumor and solid tumor model,and to study the anti-tumor activity of the drug.Through the ascites volume and the tumor size to determin the antitumor activity and dosage of administration;Experiment was conducted on immune organs and immune factors in serum mice,the immune system of the effective compounds on tumor bearing mice has a regulatory role,so we speculate that the antitumor activity the effective compounds maybe relate to the immune activity of mice.Conclusion: The antitumor activity of the effective compounds is enhanced in vivo and in vitro and the toxicity is decreased.Accordingly,we speculate that the effective compounds may be one of the important material basis of antitumor activity of cantharis metabolic liquid.