| Chapter one,Literature reviewThis chapter systematically summarizes the modern research on the effiency components group of frankincense,myrrh and their compatibility,clarifies the material basis of the traditional efficacy of frankincense-myrrh compatibility,and reviews the modern research progress of the anti-tumor activity of frankincense-myrrh compatibility.It provides a scientific basis for the development and clinical application of frankincense-myrrh.Chapter two,Preparation and analysis of the effective fraction of frankincense-myrrh compatibilityTo clarify the material basis of frankincense-myrrh compatibility in vitro and provide samples for subsequent experiments,this chapter uses liquid-mass spectrometry?UPLC-TQ/MS?methods to analyze and evaluate the 10 terpenoids in frankincense and myrrh before and after their compatibility,and optimize the preparation process of the total terpenoids of frankincense-myrrh compatibility.The results are as follows:1.The results of terpenoids analysis in frankincense and myrrh before and after their compatibility showed that the total content of 10 terpenoids in single medicinal herbs was 2.02%,wlile in the compatibility was increased to 3.40%.Except for the slightly reduced content of abietic acid,the contents of the other 9 terpenoids were all increased.The increase in the content of 3-acetyl-11-keto-?-boswellic acid,?-boswellic acid and acetyl 11?-methoxy-?-boswellic acid was most pronounced.2.The results of terpenoids extraction process optimization showed that the optimum extraction process of terpenoids was 12 times amount of 95%ethanol reflux extraction for 2times,2 h each time.Under this condition,the total content of 10 terpenoids is 1.70%.The results of terpenoids purification process optimization showed that the optimum purification process of terpenoids was 200-300 mesh silica gel,1:20 sample loading,2 BV petroleum ether:ethyl acetate 50:1 as impurity removal agent,the 4th to 6th BV petroleum ether:ethyl acetate 4:1as elution.Under this condition,the total terpenoid content was 26.60%.3.The total extracts of frankincense,myrrh and their drug pairs were prepared according to the optimized extraction and purification process,and the contents were analyzed.The results showed that the content of the total terpenoids of the frankincense ethanol extract,myrrh ethanol extract,frankincense-myrrh pair ethanol extract,frankincense water extract,myrrh water extract,frankincense-myrrh pair water extract and frankincense-myrrh pair terpenoid fraction were 8.84,11.34,17.00,0.36,1.02,0.69 and 266.11 mg·g-1,respectively,which provided the quality standard of the test samples for subsequent experiments.Chapter three,Evaluation and investigation the antitumor activity of frankincense and myrrh before and after their compatibility and its mechanismThis chapter focuses on the anti-tumor activity of frankincense and myrrh,with multiple myeloma as effect disorder,U266 and RPMI8226 multiple myeloma cell lines as research objects,combined traditional molecular biology with metabolomics techniques to explore the inhibition effects of multiple myeloma and its mechanism.It provide a scientific basis for the modern pharmacology research of frankincense-myrrh.1.U266 and RPMI8226 cell lines were used as research objects to assess the efficacy of frankincense ethanol extracts,myrrh ethanol extracts,frankincense-myrrh ethanol extracts,frankincense-myrrh total terpenoid site and and their main compounds against multiple myeloma.The results showed that the 3 extracts and frankincense-myrrh total terpenoid site significantly inhibited the proliferation of U266 and RPMI8226 cells at concentrations greater than 25?g·mL-1,3-O-acetyl-?-boswellic acid,3-acetyl-11-keto-?-boswellic acid and 11-keto-?-boswellic acid had the most significant inhibitory effect of multiple myeloma cells proliferation among the10 main compounds,and all the treated groups might play an anti-multiple myeloma effect by inhibiting the secretion of IL-6 and VEGF while inhibiting the activation of the JAK/STAT signaling pathway.2.Based on metabonomics technology to study the effects of 3 extracts and frankincense-myrrh total terpenoid site on U266 cells.The metabolic profile of U266 cells were significantly regulated by the four extracts.11,8,7,7 endogenous metabolites were identified and 2,2,3,0 related metabolic pathways were constructed in the cells of frankincense ethanol extracts group,myrrh ethanol extracts group,frankincense-myrrh ethanol extracts group,frankincense-myrrh total terpenoid site group respectively,namely vitamin metabolism,arachidonic acid metabolism,amino acid metabolism and lipid metabolism.It indicated that the effiency components group of frankincense-myrrh may play an anti-tumor effect by regulating the metabolism of U266 cells.Chapter four,Study on absorption and metabolites of the terpenoids in compatibility of frankincense and myrrh in vivoTo clarify the material basis of frankincense and myrrh in vivo,this chapter established LC-MS/MS method to analyze and evaluate the pharmacokinetics and metabolites of3-acetyl-11-keto-?-boswellicacid,11-keto-?-boswellicacidand2-methoxy-8,12-epoxygermacra-1?10?-7,11-trien-6-one before and after compatibility of frankincense and myrrh.It clarifies the absorption and metabolism mechanism of the terpenoids of frankincense and myrrh in vivo,and provides a basis for further research.The results are as follows:1.The pharmacokinetic of terpenoids in frankincense and myrrh before and after their compatibilityThe contents and pharmacokinetic parameters of 3-acetyl-11-keto-?-boswellic acid,11-keto-?-boswellic acid and 2-methoxy-8,12-epoxygermacra-1?10?-7,11-trien-6-one in plasma of rats were compared and analyzed before and after the compatibility of frankincense and myrrh,based on LC-MS technique.The results showed that 3-acetyl-11-keto-?-boswellic acid,11-keto-?-boswellic acid and 2-methoxy-8,12-epoxygermacra-1?10?-7,11-trien-6-one reached the highest plasma concentration at 2 h,2 h and 15 min,respectively.The AUC0-t-t and t1/2/2 of these three components showed different degrees of increase,while CL/F showed significant decrease.It is indicated that the compatibility of frankincense-myrrh significantly improves the absorption of the effiency components group in rats.2.The metabolites of terpenoids in frankincense and myrrh before and after their compatibilityThe metabolites of 3-acetyl-11-keto-?-boswellic acid,11-keto-?-boswellic acid and2-methoxy-8,12-epoxygermacra-1?10?-7,11-trien-6-one in plasma of rats were identified and the differences in metabolites between before and after the compatibility of frankincense and myrrh were analyzed.The results showed that there were kinds of phase I and phase II metabolites of frankincense,myrrh and the compatibility of frankincense-myrrh in vivo of the rats,and there are some differences of metabolites between single herb administration and the couplet medicines administration.The main forms of metabolites include parent,reduction,oxidation,dehydration,hydrolysis,deacetylation,dealkylation,hydroxylation,acetylation,glucuronidation,amino acid conjugates and so on. |
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