Mechanism Of Chinese Medicine Jiawei Fengfang Pill On Prevention And Treatment Of Breast Cancer

Objective:Based on the evaluation of Jiawei Fengfang Pill’s animal experiments to prevent and treat breast cancer,explore the molecular mechanism of breast cancer prevention and treatment through network pharmacological methods;artificial musk as an artificial substitute for natural musk in formulations,in order to comprehensively control the quality of preparations,a pre-column derivatization HPLC method was developed for the determination of muscone.Methods:A rat breast cancer model was constructed using7,12-dimethylbenzoanthracene(DMBA,100 mg/kg)and intervention was performed with Jiawei Fengfang Pill,observe the weight,death and tumor occurrence of rats in different intervention groups,detection of estrogen levels in peripheral blood of rats in different intervention groups;retrieve monomer compounds of 11 kinds of Chinese herbal medicines in Jiawei Fengfang Pills from TCMSP database of Chinese medicine,TCM@TANWAN of Taiwan traditional Chinese medicine database,and CNKI,uploads these monomers to the Swiss and Superpred websites to predict targets for these monomer compounds,and builds of a"Chinese herbal medicine-monomer compound-target"network;analyzes of human breast cancer transcriptome sequencing data,screens for targets related to breast cancer;intersect between the modified Hive pill targets and the breast cancer targets,gets breast targets that Modified Fengfang Pill can interfere with;analyzes the function of the above targets in the DIVID database to study the molecular mechanism of Jiawei Fengfang Pills to protect breast cancer;pre-column derivatization using 2,4-dinitrophenylhydrazine,determination of muscone content in three different extraction processes:absolute ethanol,diatomaceous earth ethyl acetate,and anhydrous sodium sulfate ethyl acetate,determine the best extraction method of muscone and test for muscone content.Results:The results of pharmacodynamic research experiments show that Chinese medicine Jiawei Fengfang Pill can affect the weight gain of breast cancer rats,protect DMBA-induced weight loss in rats,reduce DMBA-induced breast cancer rat mortality,reduce tumor diameter in breast cancer rats,regulation of estrogen and progesterone levels in experimental rats;network pharmacological analysis shows that compounds such as Bupleurum chinese DC and Scutellaria barbata D.Don in Chinese medicine Jiawei Fengfang Pill can regulate target proteins related to breast cancer,it mainly regulates the expression of CDK2,VEGFA,PLK1,MMP9 and other breast cancer-related proteins,affects p53 signaling pathway,protein serine/threonine kinase activity and other signaling pathways,regulates cell cycle,hormone levels,oxidoreductase activity and metabolic pathways,impact on breast cancer cell proliferation,metastasis,tumor neovascularization and other aspects;the pre-column derivatization HPLC test results showed that the best extraction method is anhydrous sodium sulfate ethyl acetate extraction method,the regression coefficient R~2>0.999in the determination of muscone content by DNPH derivatization method has good linearity,the recoveries of low,medium and high concentrations meet the test standards,RSD<10%,good repeatability,RSD<10%,stable at room temperature,RSD<5%,the content of muscone in different types of Chinese medicine Jiawei Fengfang Pills are 17.59μg/g,17.15μg/g,17.27μg/g,RSD<10%.Conclusion:The Chinese medicine Jiawei Fengfang Pills regulates the levels of estrogen and progesterone in breast cancer rats and has an effect on tumor bearing in rats,reduces the incidence of precancerous lesions in breast cancer,through multiple ways and multiple targets,affects breast cancer cell cycle and proliferation and invasion ability,determination of muscone content by HPLC,which is provided experimental basis for clinical application of Jiawei Fengfang Pills.