Study On Chemical Constitiutions And Antitumor Activity Of Xiaoai Jiedu Recipe

Xiaoai Jiedu recipe is an effective formula formed by long-term clinic ptactice which prescribed for cancer treatment formed by compatibly combining herbals through treatment directed by the "ai du" pathogenesis theory.It is mainly composed of Scutellariae Barbatae Herba,Akebiae Fructus,Cremastrae Pseudobulbus,Curcumae Rhizoma,Pseudostellariae Radix,Ophiopogonis Radix.In this formula,Scutellariae Barbatae Herba acts as the emperor herb with the function of clearing heat and removing toxins;Akebiae Fructus,Cremastrae Pseudobulbus and Curcumae Rhizoma are as minister herb with the effect of subduing swelling and resolving hard lumps;Pseudostellariae Radix,Ophiopogonis Radix both play the auxiliary role with the function of filling qi and nouring yin.Therefore,the prescription works together to remove toxic substance,strengthen body resistance and eliminate pathogenic factors.Most of studies on Xiaoai jiedu decoction(XAJDD)mainly focuses on the clinical practice and pharmacological studies,while the reports on chemical composition of the formula are few and no studies about metablic process of XAJDD in vivo is available.For the research of pharmaceutical material basis of XAJDD,in this paper,the chemical constituents and metabolites of XAJDD in vitro and in vivo were detected and identified by ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Finally,the anti-inflammation effect of druged serum and the anti-cancer activity of effective components were explored.This research content mainly includes the following aspects:1.Identification of chemical constituents in XAJDDAn UPLC-Q-TOF-MS method was applied to analyze the constituents of XAJDD 102 compounds were identified or tentatively characterized,including 30 flavonoids,19 triterpenoids,9 steroidal saponins,12 organic acids,9 cylic peptides,5 amino acids,7 phenanthrenes,3 alkaloids,8 other compounds.The results above provide the basis for screening of medicinal ingredients and the metablic process study in vivo.2.Characterization of the absorbed constituents and metabolitesAn UHPLC-Q-TOF-MS method was established to analysis the absorbed constituents in doesed rat plasm and urine for the first time in this paper.As the results,15 prototypes(includes 9 flavonoids)and 53 metabolites transformed from these prototypes were identificated.Based the results above,the metabolic pathways of them was built.Finally,the distribution of the metabolites in eight rat organs were reported.Therefore,The metabolic mechanism of absorbed constituents in XAJDD and distribution of the metabolites were preliminary explored.3.Evaluation of the anti-in flammmation activity of druged serumLPS-induced rat macrophage RAW264.7 model was used to evaluate anti-inflammation activity of different concentration of Xiaoai Jiedu prescription medicated serum and NO production was used as a detection index.The results showed that:compared with the control group,the medicated serum had no effect on the valilty of RAW264.7 cell in the concentration range from 2%to 6%;compared with the LPS group,the concentration of the medicated serum was at the range from 2%to 6%,which showed a certain efffect on NO emissions,and the difference was statistically significant.Therefore,we indicated that the medicated serum has certain anti-inflammtory activity in this concentration range,which may be related to the flavonoid as the main absorbed component.4.Screening of effective components with anti-cancer activity in XAJDDThe anti-cancer ability of the effective components in XAJDD was studied against Gastric cancer and colorectal cancer cell lines respectively.The results showed that:compared with control group,when the concentration of each compound was 30ug/mL,the compound α-hederin showed significant activity inhibition on the activity of all the cancer cell lines tested,indicating that the tumor inhibiting activity of it was remarkable.In addition,when the concentration ofα-hederin was 15 ug/mL,SW620 cell was the most sentitive cell amone all the cancer cell lines tested,and the SW620 cell viablitty decreased most signigicantly after treated.5.Anti-cancer effect and mechanism of α-hederinThe study was aimed to evaluate the potential antitumor activity of α-hederin in human colorectal cancer cells stimulated with interleukin 6(IL-6)for micicking the tumor inflammatory microenvironment in vivo.The results showed that:IL-6 at 6.25 ng/mL significantly enhanced viablility,migration and invasion of SW620 cells,but α-hederin inhibited cell viability,migration and invasion in IL-6-treated SW620 cells and restored abnormal expression of EMT markers.Moreover,α-hederin inhibited phosphorylation of JAK2/STAT3 and halted the nuclear translocation of phosphorylated STAT3.Finally,it was found that α-hederin has synergistic effect with AG490.SO we demonstrated that α-hederin suppressed IL-6 induced Epithelial to mesenchymal transition(EMT)associated with disruption of JAK2/STAT3 signaling in colorectal cancer cells.