Study On Material Basis Of Eclipta Prostrata L. In The Prevention And Treatment Of Lung Cancer Based On Component Structure Theory

Lung cancer is one of the most threatening malignant tumours to human health. The combination of prevention and treatment are commonly taken to reduce the morbidity and mortality of lung cancer for a long time. Traditional Chinese medicine has become an important drug sources of tumor prevention and treatment due to its unique advantages in this area. Eclipta Prostrata L. is one herb of the "Three grass" formula as an antitumor compound, but few reports have been made on it as a single herb antitumor.In this dissertation, the prevention and therapy effects of Eclipta Prostrata L. for lung cancer were evacuated with different models. The active materials basis of Eclipta Prostrata L.were selected and optimized based on the "component structure" theory. The oral bioavailability was improved by the means of pharmaceutical formulation for those active components with poor absorption. The main contents are as follows:1. The effects evacuation of Eclipta Prostrata L. on the prevention and treatment of lung cancerThe tumor inhibition effects of EEP (Extract of Eclipta Prostrata L.) extracted from different regions were evacuated and proved using C57mice vaccinated Lewis cells. The tumor inhibition rate of EEP reached the maximum of45.21±4.62%with the crude drug dosage of10g/kg/day.A549cell model was used to evaluate the anti-lung cancer activity of the EEP. The results show that the EEP has inhibitory effect on A549cell.The value of IC50is308μg (crude drugs)/mL. The cell apoptosis rate of A549reached to47.55%after24hours when the EEP concentration of crude drugs is1000μg/mL, verifying the cell apoptosis effect of EEP on A549cells. After be treated with EEP the amount of pro-apoptotic protein in A549such as Bax, cleaved-caspase9, cleaved-caspase3and cleaved-caspase3increased, while the amount of the inhibitor of apoptosis Bcl-2decreased.The CSE (Cigarette smoke extract) induced NHBE cells model was built up to evacuate the protection effect of EEP on the normal pulmonary epithelial cells. The survival rate of the CSE induced NHBE cells increased apparently with the addition of EEP in dose-dependly. The antioxidant index shows significant activity reduction of SOD、CAT and GSH in the CSE induced NHBE cells, and the amount of MDA increased. Both the level of SOD, CAT, GSH and the content of MDA were adjusted to normal values with the use of EEP. The protein expression levels of inflammatory markers COX-2and ICAM-1(measured by Western blot) increased obviously in NHBE cells after treated with CSE; Interestingly, both of the values decreased and approached to normal value after EEP treatment, which indicate that EEP plays an certain role in of the chemistry induced lung cancer. The evacuation results of the models discussed above confirmed the positive effect of EEP on the prevention of lung cancer.2. The separation and enrichment of the EEP componentsThe EEP components were separated and enriched by using automatic preparation system and macroporous resin. UPLC/Q-TOF MS was used to check the index components. For the first time, isoquercitrin, isochlorogenic acid C and isochlorogenic acid A were found in Eclipta Prostrata L.. The quantity analysis results on t enriched components showed that phenolic acids, flavonoids, coumestans and triterpenoids components were successfully collected.3. The structural optimization on EEP componentsThe structural optimization on EEP components was made combing the A549cell model and cigarette induced NHBE cell model. All the four components phenolic acids, flavonoids, coumestans and triterpenoids in EEP contribute to pharmacodynamic effect. They showed strong inhibiting effects on A549cell and prevention effects on cigarette induced NHBE cell. The optimized structural ratio are11:9:10:18～22..To verify the inhibiting effects of the optimized EEP components structure, C57mice inoculated with Lewis cells were used. Based on the above results, the optimized structure of the components in Eclipta Prostrata L. was set as phenolic acids:flavonoids:coumestans: triterpenoids=11:9:10:18.4. Enhanced bioavailability of flavone components in EEP via preparation of solid dispersion using carbon nanopowdersFirstly, carbon nanopowders(CNPs) was used in pure apigenin(AP). Solid dispersions of AP with CNPs were prepared with the ratio of1:6. Drug-release profiles showed that AP dissolution from the CNP-AP system (weight ratio,6:1) after60minutes was improved by275%as compared to the pure drug. Moreover, pharmacokinetic analysis of the solid dispersion formulations in rats showed that the AP area under the curve0-t value was1.83times higher for the CNP-AP system as compared to pure AP, indicating that bioavailability was significantly improved. Additionally, the solid dispersions had a significantly higher peak and shorter time-to-peak as compared to pure AP. Preliminary intestinal toxicity tests indicated that there was no significant difference in the tissues of CNP-AP system-treated, CNP-treated, and control rats. In conclusion, the CNP-based solid dispersions could be used for enhancing the bioavailability of flavanoids.. The solid dispersion of flavone components in EEP was prepared using CNPs as carrier. The dissolutions in vitro of luteolin and apigenin in flavone components were greatly improved, and their oral bioavailability increased by192%and176%, respectively.