Angelica Extracts From Supercritical And Fractions For Colorectal Cancer Prevention Research

Objective:The purpose of this study was to investigate the effect of CO2Supercritical Fluid Extracted (SFE) of Angelica sinettsis(AS) and its distillates on the prevention of colorectal tumorigenesis. The anti-inflammatory and anti-oxidative stress activities of SFE of AS and its distillates were investigated using in vitro approaches. Additionally, the role of the transcriptional factor Nrf2(nuclear erythroid-related factor2) signaling pathways was examined. Two chemical reagents, azoxymethane(AOM)and dextran sulfate sodium salt(DSS) were used to induce colorectal tumorigenesis. Then we investigated the effect of SFE of AS and its distillates on the prevention of cancer, and compared with vitro results.Methods:We investigated whether SFE of AS and its distillates could induce Nrf2-ARE and anti-oxidative stress activities in HepG2-C8cells with stably transfected ARE luciferase reporter genes. Stimulating HepG2-ARE6h after administration medicines, Luciferase reporter and RT-PCR assay were used to test the anti-oxidative stress activities of SFE and its distillates. LPS-challenged RAW264.7macrophagus model was used to eatablish inflammation model. The level of NO was detected by Griess reagent method. Inflammation-related gene expression of COX-2and iNOS were determined by RT-PCR method. The ability of SFE of AS and its distillates on the effect of cancer prevention were investigated by colorectal tumorigenesis in BALB/c mice induced with AOM/DSS. The mice were randomly divided into6groups:normal control group, model group(AOM+DSS), SFE of AS high dose group(AOM+DSS+60mg/kg), SFE of AS low dose group(AOM+DSS+15mg/kg), distillate4high dose group(AOM+DSS+60mg/kg), distillate4low dose group(AOM+DSS+15mg/kg). Body weight was recorded every week. After17weeks, ether anesthetized mice, the blood was collected. The level of TNF-α、SOD、Nrf2、NF-kB were detected by ELISA. Colorectum was collected, tumor number, tumor incidence and tumor location were recorded. Pathological changes were observed through HE staining.Results:(1) Cell experiments:SFE of AS and its distillates can dose-dependently activated ARE luciferase. Distillate4performed best effect on activating Nrf2、HO-1、NQO1expression, showed the best antioxidant. SFE of AS and all the distillates can inhibited NO， and the SFE ofAS、Distillate4and Distillate5showed best. SFE of AS>Distillate4=istillate5>others. Distillates4、5、6showed better inhibition on the level of COX-2、iNOS than SFE of AS.(2) In vivo:After drinking DSS, mice were found bloody stools, diarrhea, few mice rectal prolapse.normal control group found no tumor, but the incidence of model、、 group was100%, High/Low dose SFE of AS group was87.5%. High dose distillate4group was75%, Low dose distillate4group was62.5%. High dose distliiate4group showed best effect on inhibition TNF-α. NF-kB and increase Nrf2、SOD.Conclusions:(1)Angelica liposoluble constituents can play the role of the anti-inflammatory and antioxidan by activating Nrf2/ARE pathway. Distillate4performed the most significant effect.(2) Angelica liposoluble constituents can inhibit the development of colorectal cancer, and distillate4was found best.This shows that lipids in SFE of AS have a synergistic action for cancer prevention, whichisworthfurtherresearch...