| Balsam pear(Momordica charantia L.)is the dried and nearly mature fruit of Momordica charantia Linnaeus,also known as litchi,madness grape,cold melon and so on.It is a traditional Chinese medicine which is homologous to food and medicine.The Chinese Dictionary records that the roots,stems,leaves,flowers,fruits and seeds of balsam pear can be used as medicine,while the Compendium of Materia Medica records that they can relieve fatigue,eliminate evil fever and clear the mind.Taste bitter,cold,return to the heart,spleen,liver meridian,can clear heat,clear eyes,detoxification.It is often used to treat summer heat,thirst,red eye pain,dysentery,intestinal carbuncle and so on.A large number of literature studies have shown that balsam pear extract can effectively inhibit the growth of liver cancer cells,but the material basis of its anti-liver cancer effect has not been systematically reported.In this paper,on the basis of previous literature research,through the interdisciplinary research method,the separation and purification of Momordica charantia alcohol extract and the analysis of effective components were carried out,and the antihepatoma mechanism of the active components was studied.OBJECTIVE: To interpret the pharmacodynamic material basis and action mechanism of bitter melon alcohol extract against liver cancer,provide a modern scientific basis for its clinical efficacy against liver cancer,and lay a foundation for the development of innovative traditional Chinese medicine or antihepatocellular health food.METHODS: Fresh balsam pear was extracted with ethanol and precipitated with acetone to get the alcohol extract of Momordica charantia.Taking the content of effective substances in balsam pear as an index,the best macroporous adsorption resin was selected for the preliminary purification of balsam pear alcohol extract,and the effective components of of Momordica charantia alcohol extract and macroporous resin purified part were isolated and identified by modern HPLC-MS analysis method.The inhibition of balsam pear alcohol extract and macroporous resin purification site on liver cancer cells was verified by H22 tumor bearing mice as animal model,and modern molecular biology technology was adopted to study the action mechanism of the components with anti-hepatoma activity.At last,it was further separated and purified by preparative high-performance liquid chromatography.Meanwhile,the separated components were also analyzed and identified by HPLC-MS,and the peptide library with hepatoma activity was established.RESULTS: The content of alcohol extract of Momordica charantia was determined.The results showed that the highest content of the effective components in alcohol extract was polypeptides,and the second was saponins.Therefore,taking the content of polypeptides and desalination rate as index,we screened out DA201-C macroporous adsorption resin for preliminary purification.The content of polypeptides purified by macroporous adsorption resin reached more than 50%,while the content of saponins was also slightly increased,and the content of saponins was 8.1%,but the increase was not significant.The anti-tumor experiments of Momordica charantia alcohol extract and macroporous adsorption resin on H22 tumor-bearing mice showed that both of them had certain anti-tumor effect(P < 0.05).The tumor inhibition rate of the purified part of Momordica charantia was significantly higher than that of alcohol extract(P < 0.05).The polypeptides from the macroporous adsorption resin were analyzed and identified by HPLC-MS.A total of 86 peptides from 62 proteins of Momordica charantia were detected.The study on the mechanism of action showed that the purified fraction of macroporous adsorption resin could enhance the expression of Bax and caspase3 and decrease the expression of Bcl-2,p-STAT3 and PD-L1.The experiment of serum Elisa also showed that the purified fraction of macroporous adsorption resin could increase the level of serum TNF-? and IL-2 and decrease the level of IL-6 in H22 tumor-bearing mice.Six components(P1-P6)were further isolated and purified by high performance liquid chromatography(HPLC).All of them were polypeptides determined by BCA kit,and the content was more than 98%.Unfortunately,these six components are not a single component,and the next step of separation and purification will be carried out around this step.CONCLUSION: Based on the comprehensive analysis of the research results of this paper,it is found that both the alcohol extract of Momordica charantia and the purified part of macroporous adsorption resin can effectively inhibit the growth of tumor in H22 tumor-bearing mice,promote the apoptosis of tumor cells in tumor-bearing mice and play the role of antiliver cancer.This study also found that the purified part of macroporous adsorption resin could inhibit the proliferation of H22 hepatoma cells through Caspase-3/Bax/Bcl-2 signal pathway,enhance TNF-?,IL-2,and decrease the expression of IL-6 and PD-L1,so as to further promote the apoptosis of tumor cells.Through the study of the anti-hepatoma active components of the alcohol extract of Momordica charantia,it was found that the substance that could promote the apoptosis of tumor cells were polypeptides.This paper mainly discussed the material basis and mechanism of Momordica charantia alcohol extract on antiliver cancer,which laid a foundation for the development of innovative drugs or health food of balsam pear traditional Chinese medicine. |
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