Study The Effects Of Total Phenols And Flavonoids In Compound Capillus-veneris Granules On Cervical Cancer Cells Based On Cell Metabonomics

Objective: To investigate the effects of total phenol and total flavonoids in compound Adiantum capillus-veneris granules and study its effect on cervical cancer cells such as SiHa,HeLa and C-33 A.Based on 1H-NMR,the metabolic pathways related to proliferation and apoptosis of cervical cancer cells were analyzed.Methods: Total flavonoids and polyphenol extracts were extracted and purified from the compound ferrogonium fern by extraction process.The standard curves of rutin and gallic acid were prepared and determined by ultraviolet spectrophotometry.The total flavonoids and total phenols of different concentrations were given to 3 kinds of cells,and the IC50 values of each cell under the action of 3 different extracts were determined by a drug cytotoxicity test.Take 1/2 times IC50,IC50,2 times IC50 as low,medium and high concentration,and apoptosis was detected by FITC-conjugated Annexin-V staining;Cell culture medium and cell samples of normal control group,total phenol group(low,medium,high),total flavone group(low,medium,high),and positive drug group(cisplatin 25 ?g / mL)of cervical cancer cells were collected.,Cell culture medium and cell samples of 24 groups were detected by 1H-NMR.Multivariate statistical analysis methods such as partial least squares discriminant analysis(PLS-DA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to find differential metabolic pathways related to cancer cell proliferation and apoptosis.Results: The total phenol content in compound Adiantum granules was 30.83%,and the total flavonoid content was 39.14%.Compared with the normal control group,the SiHa,HeLa and C-33 A cells with different concentrations of the two extracts increased the inhibition rate with the increase of the concentration(P <0.01).The apoptotic rate of the 3 kinds of cells under the action of total phenol and total flavonoids was significantly different from that of the normal control group,and the difference between the total phenol-administered group and the total flavonoid-administered group was significant.In the OPLA-DA diagram of the two groups,we can know that the two groups are well separated,which shows that there are differences in metabolites between the two groups.Among them,there are 35 different metabolites involved in HeLa cell culture media and cells: isoleucine,leucine,valine,lactic acid,alanine,lysine,acetate,proline,glutamine,Carnitine,glutamic acid,dimethylamine,aspartyl,glutamate,a-Ketoglutaric acid,creatine,choline,taurine,glycerylcholine,?-glucose,aconitic acid,hydrazone-glucose,Glycine,guanidinoacetate,3,4-dihydroxymandelate,tyrosine,uronate,phenylalanine,adenosine phosphate,3-hydroxybutyrate,sarcosine,trimethylamine,?-furanose,ascorbic acid,Hippurate,3-methylhistidine;There are 37 different metabolites in SiHa cell culture media and cell types: isoleucine,leucine,valine,lactic acid,alanine,lysine,acetate,proline,glutamine,Acetoacetic acid,carnitine,glutamic acid,sarcosine,dimethylglycine,methylguanidine,trimethylamine,aspartyl,creatine,citrulline,choline,glycerophosphocholine,?-glucose,?-glucose,glycine,?-furanose,inositol,ascorbic acid,3,4-dihydroxymandelate,tyrosine,uronate,phenylalanine,3-methylhistidine,adenylate,adenosine phosphate,Dimethylamine,sarcosine,trimethylamine-N-oxide;There are 34 different metabolites in C-33 A cell culture medium and cells: isoleucine,leucine,valine,lactic acid,alanine,lysine,acetate,proline,and glutamine,Carnitine,methylamine,sarcosine,a-Ketoglutaric acid,citrulline,choline,taurine,TMAO,glycerophosphocholine,beta-glucose,beta-glucose,glycine,beta-furanose,Ascorbic acid,creatine,inositol,3,4-dihydroxymandelate,tyrosine,uronate,phenylalanine,3-methylhistidine,adenosine phosphate,formic acid,albumin lysyl,trans-Aconitic acid.These differential metabolism are involved in many metabolic pathways.Conclusion: Analysis of the metabolic pathways of each cell culture medium: phenylalanine,tyrosine and tryptophan biosynthesis,taurine and hypotaurine metabolism,alanine,aspartic acid and glutamic acid metabolism,D-Glutamine and D-glutamic acid metabolism,phenylalanine metabolism,arginine biosynthesis,glycine,serine and threonine metabolism,arginine and proline metabolism,histidine metabolism,glutathione Glycine metabolism,glyoxylic acid and dicarboxylic acid metabolism,tyrosine metabolism and other metabolisms have changed;the metabolism involved in different metabolites of various cells includes: phenylalanine,tyrosine and tryptophan biosynthesis,Synthesis and degradation of ketone bodies,taurine and sulfinate metabolism,phenylalanine metabolism,glycine,serine and threonine metabolism,arginine biosynthesis,alanine,aspartic acid and glutamine Acid metabolism,tyrosine metabolism,histidine metabolism,butyrate metabolism,glyoxylic acid and dicarboxylic acid metabolism,arginine and proline metabolism,D-glutamine andD-glutamic acid metabolism,Glutathione metabolism,cysteine?and methionine metabolism,pyruvate metabolism,glycolysis / gluconeogenesis,etc.Total phenol and total flavonoid can inhibit the proliferation and induce apoptosis of SiHa,HeLa and c-33 a cells through energy metabolism,amino acid metabolism,lipid metabolism and nucleic acid metabolism.It will lay a foundation for the application of cell metabonomics to interpret the metabolic pathway of tumor cells more accurately at the cellular level.