Composition Identification And Protective Mechanism On Radiation Induced Oxidative Damage Of Lonicera Caerulea Var. Edulis Anthocynin

Ionizing radiation can induce acute or chronic damage in the nervous,hematopoietic, digestive and immune systems through reactive oxygen species (ROS)caused oxidative stress in cells and tissue. Hence screening and evaluation antioxidantand radiation protective activity components from natural product resources havebecome a hot spot in the study of biology, medicine and food science. In this thesis,separation, purification and identification of anthocyanin in Lonicera.caeruleavar.edulis (ALC) were researched; antioxidant activity and protective effect onradiation-induced oxidative damage of ALC were also investigated. The main results areas follows:The optimum extraction conditions for ALC were47%ethanol,97min extractiontime, and the ratio of material to solvent of1:16.35. The optimum purificationconditions for ALC were AB-8macroporous resin,4.0BVs ethanol (50%, pH=3.0)eluant under1.5BV/h flow rate, and the ratio of diameter to height of1:15. Then we got22.26purification multiple than crude extracts and79.68color value ALC.ALC constituents were analyzed and identified by HPLC-DAD, HPLC-ESI-MS, Q-TOF-MS, UV-VIS and IR. The results show that the main constituents ofALC contained eight anthocyanins: cyaniding-3-sophoroside-5-glucoside, cyanidin-3,5-diglucoside, cyaniding-3-(feruloyl)-glucoside, cyaniding-3-glucoside, cyanidin-3-rutinoside, pelargonidin-3-glucoside, peonidin-3-glucoside and peonidin-3-rutinoside.Their mass percent contents were1.12%,3.71%,1.38%,72.42%,9.62%,2.30%,7.10%and2.34%, respectively.Antioxidant activities of ALC were studied in vitro, such as removal physiologicaland non-physiological free radicals, total reducing capacity and anti lipid peroxidationactivity. The results show that ALC has high scavenging activity on hydroxide radical(·OH). The main mechanism is that ALC is an active hydrogen atom donor, which canbreak down radical chain reactions through hydrogen atom transfer. ALC also showsignificant total reducing capacity and inhibition activity on lipid peroxidation with adose-dependent manner.X-ray or60Coγ radiation induced mouse spleen cell damage model was establishedto test and verify ALC’s significant protective effect on radiation induced damagethrough enhancing cell viability and reducing DNA damage induced by ionizingradiation.Then, oxidative damage animal model induced by60Coγ radiation was establishedto investigate the protective effect in vivo. The results show that ALC can significantlyreduce mouse immune organs oxidative damage through enhancing antioxidant enzymes activities such as superoxide dismutase (SOD) and glutathione peroxidase(GSH-Px). ALC can ameliorate glutathione (GSH) and reduce malondialdehyde (MDA)level in organs and serum of mouse to decrease lipid peroxidation and protect cellmembrane. ALC also can inhibit generation of atypical lymphocytes and bone marrowmicronuclei induced by radiation.Regulating effect of ALC on apoptosis and MAPK path related proteins expressstatus were analyzed through western blot assay. The results demonstrate that theprotective effect mechanism of ALC on oxidative damage induced by radiation was thatALC can increase expression of Bcl-2and HSP70, block radiation-induced up regulatedof Bax, phosphorylated JNK1/2and p38, inhibit the release of cytochrome c, depressthe activities of caspase-3, and thus inhibit cell apoptosis.These results demonstrate that ALC has effective antioxidant and antiradiationactivities through regulating cell oxidoreduction and apoptosis signal transduction. Thepresent study has important theoretical and practical significance to revealingmechanism of preventing radiation induced oxidative damage and developing newradioprotectors.