The Study Of Sanguinarine On Antioxidant Activity And MKN-45and TE-1Cancer Cells Apoptosis

Reactive oxygen species (ROS) is normal metabolic product of aerobic organisms，it canmaintain the body’s physiological function to achieve dynamic balance. When in the stressstate, the organisms produce excess ROS which causes oxidative damage to the body, such asprotein oxidation, lipid peroxidation and DNA oxidative damage, etc. In order to reduce thedamage of excessive oxidative to the body, some natural antioxidants gradually attract theattention of the researchers. Sanguinarine(SAN), a quaternary benzylisoquinoline alkaloids,mainly exists in the poppy family, fumariaceae and rutaceae, but at the moment, there is littlereport to the in vitro antioxidant activity of SAN.The purpose of this paper is to evaluate antioxidant activity of SAN in vitro, providing atheoretical basis for SAN as a natural antioxidant application. The scavenging capability ofDPPH radical was measured by graded levels of SAN;To investigate the protective effect ofSAN on oxidative and carbonylation damage, the BSA oxidative and carbonylation damagewas induced by Cu2+/H2O2and AAPH system, respectively; TBARS method was applied todetermine the inhibition of the soybean lecithin and herring sperm DNA oxidative damageinduced by AAPH system by SAN. The results showed that SAN can effectively removeDPPH free radicals in a dose-dependent manner, the IC50was47.09μmol/L, or15.65mg/L;Adding1to100μmol/L SAN could remarkably protect BSA protein damage induced byCu2+/H2O2and AAPH system. Moreover, SAN could considerably inhibited FeSO4inducedlipid peroxidation, as well as the oxidative DNA damage induced by AAPH.100μmol/LSAN on lipid peroxidation and oxidative DNA damage inhibition rate were84.92%and62.73%, respectively. The data which have been motioned proved that SAN has its owncapability as natural antioxidants.The researches have shown that SAN could induce apoptosis effects on large rangekinds of cancer cells, however, for SAN could induce apoptosis in cancer cells which in thedigestive system compares the molecular mechanisms have been barely reported. The targetcancer cells in the research were MKN-45human gastric cancer cells and human esophagealcancer cells TE-1, in order to deal with SAN as a factor for the trial of the two cell viability byMTT assay, AO/EB and DAPI fluorescence staining of cell morphology changes, and explore the JC-1fluorescence probe changes in mitochondrial membrane potential and the use of FasFasL and caspase-8protein to establish a protein receptor-related pathway. Furthermore,apoptosis was measured by the Western blot extrinsic pathway of apoptosis death rants tocompare the apoptotic effect of SAN on MKN-45cells and TE-1cell-associated protein, forinstance, Bcl-2, Bax, PARP, Caspase-3and cyto-c pathway. The results indicated that SANcould decrease the ratio of the Bcl-2/Bax, in addition, SAN could activate Caspase-3PARP aswell to release cyto-c into the cytoplasm. The outcome demonstrated that SAN has thecapability to promote the apoptosis of MKN-45cells and TE-1cells by mitochondriaapoptosis pathway, in which, the apoptosis of MKN-45cells revealed more remarkable thanwhich in TE-1cells. Alternatively, the SAN could promote the apoptosis of MKN-45cells bythe foreign signal of Fas receptor-mediated pathway, while TE-1cells would not affected bythis pathway.The most current research was based on previous researches illustrated that whether thePI3K/Akt and MAPK signaling pathway is involved in the process of inducing apoptosis bySAN. The outcomes showed that they both involved in apoptosis of both kinds of cells. Forthe MAPK signaling pathway, MKN-45cells is regulated by MAPK pathways associatedprotein, for instance, p-p38, p-JNK and p-ERK. Meanwhile, the p-ERK protein is notinvolved in TE-1cell regulation which effected by p-p38and p-JNK as well.This research shows SAN has the function of removing DPPH free radicals and inhibitingoxidation of biological macromolecules, meanwhile it can promote cell apoptosis of humangastric cancer MKN-45cells and Esophageal cancer TE-1cells, the above makes a theoreticalfoundation for SAN to be as a natural antioxidant and an anti-cancer health care product.