| Purpose:The LC-MS/MS analysis technique was used to explore the differences of pharmacokinetics in rat plasma and distribution of mouse tissues before and after the combination of hydroxylcamptothecin and"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma".And study on the pharmacodynamic effects of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin on lewis lung cancer-bearing mice was united in wedlock.To clarify the role of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"enhanced the targeted delivery and anti-tumor effect of hydroxycamptothecin in the lung,in order to provide a new strategy for the development of clinical treatment programs for lung cancer.Method:?1?First,LC-MS/MS analysis method of hydroxycamptothecin was established.SD rats were used as research objects,and they were divided into two groups.After administration of hydroxycamptothecin alone and in combination with"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma",the plasma was collected at different times and sampled.The content of hydroxycamptothecin in plasma was determined by LC-MS/MS.The pharmacokinetics feature of hydroxycamptothecin combined with"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"in rats before and after treatment were analyzed.?2?C57BL/6 mice were used as the research object.After administration of hydroxycamptothecin alone and in combination with"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma",the tissues?heart,liver,spleen,lung and kidney?were collected at different points in time.After tissue sample handling,LC-MS/MS technique was used to determine the content of hydroxycamptothecin in different tissues.?3?The mouse model of Lewis lung cancer was established by using C57BL/6 mice as the research object.The mice that were successfully modeled were randomly divided into model group,hydroxycamptothecin alone group and the"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"low,medium and high doses combined with the hydroxycamptothecin group.During the administration,the mice were weighed daily,and the mouse calipers were measured with vernier calipers on the 1st,4th,7th,11th,13th and 15th day after administration.The tumor volume was calculated.After 16 days of administration,the mouse eye blood was taken for blood routine measurement,and using an ELISA kit was used to determine the content of the serum IL-2,IFN-?,and IL-10 levels.The tumor tissue was removed,weighed and the tumor inhibition rate were calculated.The mouse heart,liver,spleen,lung and kidney tissue were taken to calculate the mouse organ index.Result:?1?The LC-MS/MS quantitative analysis method for the determination of hydroxycamptothecin in rat plasma established in this experiment showed that no hydroxycamp tree was detected in the rat blank plasma without hydroxycamptothecin standard solution and internal standard solution.Hydroxycamptothecin and internal standard were well separated,hydroxycamptothecin had good linearity in the range of1.362728ng/ml,the recovery rate and matrix effect were above 80%,intra-day and inter-day precision?RSD?range were 0.723.77%,the accuracy?RE?range was6.8310.29%,and the stability?RSD?range was 2.257.60%.Pharmacokinetic experiments showed that the pharmacokinetic parameters of AUC?0-t??1176.268±343.014?ug/L*h and Cmax?2604.4±865.009?ug/L of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin increased compared with those of AUC?0-t??926.356±231.58?ug/L*h and Cmax?2139.2±668.692?ug/L,but there was no statistical difference?P>0.05?.?2?The methodological results of LC-MS/MS method for determining the content of hydroxycamptothecin in C57BL/6 mouse tissues established in this chapter show that the specificity is good.There is no interference peakof hydroxycamptothecin and the internal standard camptothecin in each tissue.The linearity of each tissue standard curve was good in a certain linear range,the recovery rate and matrix effect were more than 75%,and the intra-day and inter-day precision?RSD?range were 1.2%9.58%,accuracy?RE?ranged from-13.4%to 11.45%,and stability?RSD?ranged from 0.47%to 12.56%.The pharmacokinetic parameters of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin showed that AUC?0-t?in lung tissue increased by 2.41 times and peak concentration in lung tissue increased by about 1.41 times,AUC?0-t?in heart tissue increased by 1.76 times,AUC?0-t?in liver tissue increased by 1.42 times,and AUC?0-t?in spleen tissue increased by about 1.99 times compared with hydroxycamptothecin alone.The CLz of the heart,liver,spleen and lung tissues in the combined group was significantly lower than that in the HCPT alone group?P<0.01?.?3?The anti-lung cancer effect of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin showed that on the 16th day of administration,the anti-tumor rate of hydroxycamptothecin alone group was 32.93%,while that of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"at low,medium and high doses combined with hydroxycamptothecin were 33.79%,47.14%and 50.36%,respectively;and"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin had some influence on the body weight of mice and liver index.Routine blood samples from mice showed that compared with the single chemotherapy group of hydroxycamptothecin,the high dose of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin significantly increased the number of red blood cells,hemoglobin and platelets?P<0.05?.The results of ELISA kit showed that compared with HCPT alone,the medium dose of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"was combined with hydroxycamptothecin and the high dose of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin in the serum of mice,the content of IL-2 was significantly increased?P<0.01?,while the content of IL-10 was significantly decreased?P<0.01?.The high dose of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxycamptothecin in the serum of mice,the content of-?was significantly increased?P<0.01?.Conclusion:?1?The established LC-MS/MS quantitative analysis method for the determination of Hydroxycamptothecin in rat plasma is reliable,sensitive,rapid and accurate,and can be applied to the pharmacokinetic study and concentration determination of Hydroxycamptothecin in rat plasma.Pharmacokinetic experiments showed that"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"could increase the concentration of Hydroxycamptothecin in rat plasma,and"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"could affect the pharmacokinetic behavior of Hydroxycamptothecin in rat plasma.?2?The established LC-MS/MS quantitative analysis method for the determination of Hydroxycamptothecin in mouse tissues meets the requirements of biological sample detection and can be used to detect the content of Hydroxycamptothecin in mouse tissues.The results of tissue distribution of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxy-camptothecin and hydroxycamptothecin alone in mice showed that"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"significantly increased the concentration of hydroxycamptothecin in heart,lung,liver and spleen tissues,and prolonged the elimination process of hydroxycamptothecin in heart,lung,liver and spleen tissues,indicating that"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"may not only enhance the distribution of hydroxycamptothecin in the lungs,but also enhance the distribution of hydroxycamptothecin in the heart,liver and spleen.?3?The mice were selected from C57BL/6 inbred mice to establish a lewis lung cancer tumor-bearing mouse model.The anti-tumor effect of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with hydroxy-camptothecin injection was investigated.The results showed that"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"could increase the the inhibitory rate of hydroxycamptothecin.It may be related to the decreased expression of IL-10,increased expression of IL-2 and IFN-?,significantly increased levels of red blood cell,hemoglobin and platelet,enhanced the body immunity system and decreased toxic and side effects of drugs after combined administration.In summary,"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"combined with anticancer drugs can improve the distribution and immunity of anticancer drugs in diseased tissues.The results of this study can provide a new strategy for the development of clinical treatment programs for anticancer drugs,and provide a reference for the clinical promotion of the combined application of"Platycodonis Radix and Glycyrrhizae Radix Et Rhizoma"and anticancer drugs. |
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