Effects Of Dietary Acrylamide Exposure On The Nervous System And Protective Effect Of Lycium Ruthenicum Polyphenols

Objective Dietary exposure to ACR has become a global dietary safety issue.Studies have shown that ACR has neurotoxic effects,but its mechanism is unclear,and effective preventive measures cannot be taken to inhibit the toxic effects of ACR exposure.Therefore,the study aims to understand the nerve damage caused by ACR exposure in the diet and the protective effect of Lycium ruthenicum polyphenols?LRP?on ACR-induced neurotoxicity,so as to provide a new research idea for the prevention of food safety problems caused by dietary contamination exposure.Methods 1.To study the effects of dietary ACR exposure on nerve injury in college students,food frequency questionnaire was used to investigate the diet of college students,combined with the core neurobehavioral core test battle?NCTB?and the detection of oxidative stress index in serum.2.To investigate whether has the protective effects of oxidation resistance,resistance to apoptosis,PC12 cells were used to establish H₂O₂ oxidative stress model.MTT method was used to detect cell viability,DCFH-DA fluorescence probe to detect reactive oxygen species?ROS?level,JC-1 fluorescence probe detection of mitochondrial membrane potential,Annexin V-FITC/PI staining to detect cell apoptosis,and caspase 3,caspase 8 and caspase 9 were used to detect the activation level of apoptosis factors.3.SH-SY5Y cells were used to establish ACR neural injury model,MTT method was used to detect cell viability,DCFH-DA fluorescence probe to detect ROS level,Hoechst 33258 staining to detection cell apoptosis,the levels of SOD,glutathione and MDA were measured.And Western blot method to detect the Nrf2pathway downstream antioxidants GCLC,GCLM and HO-1 level,studying whether LRP can play a role of the antioxidant of ACR nerve damage caused by the protective.Results 1.The FFQ results showed that the average exposure value of dietary ACR of college students was 0.515?g/?kg BW·day?,the median was 0.376?g/?kg BW·day?.There was no significant gender difference in the exposure,but the individual difference level of dietary ACR exposure was large.The main source of dietary ACR exposure was cereal,followed by snacks such as crisps and nuts.NCTB test showed that with the increase of ACR exposure,the abnormal emotions of the subjects increased and their behavioral functions were normal.Biochemical indexes showed that the oxidative stress level was normal.2.Utilization of PC12 cells for establishing a cell model.It was found that H₂O₂ inhibited the activity of PC12 cells significantly and increased the level of ROS in PC12 cells.The PC12 cells with H₂O₂ affects the normal function of mitochondria,which leads to apoptosis.The cell viability was improved after with LRP pretreatment intervention.Mitochondrial function was improved with the ROS level was inhibited.The apoptosis factors caspase 3,caspase 8,caspase 9induced by H₂O₂ were decreased.Studies have shown that LRP has antioxidant effect,which can protect normal mitochondrial function and inhibit cell apoptosis.3.Utilization of SH-SY5Y cells for establishing a cell model.The result found that ACR inhibited the activity of SH-SY5Y cells,ROS level increased,MDA level increased,SOD and GSH content decreased,which indicated that oxidative stress caused cell apoptosis.The SH-SY5Y cells with LRP pretreatment intervention improve cell vitality,antioxidant and anti-apoptotic effects were exerted to inhibited ROS level and reduce oxidative stress.Western blots results indicated that LRP up-regulated the expression of antioxidant protein GCLC,GCLM and HO-1.Conclusion 1.In the daily diet of college students,the snack food with high ACR content,such as potato food and nut food,easy to cause high dietary ACR exposure level.Although the ACR exposure in the daily diet does not cause obvious health harm to the nervous system,it can still cause negative effects such as the increase of abnormal emotions.Result shows that attention should be paid to the dietary safety of college students with high dietary ACR exposure level,and it is an important measure to reduce the dietary ACR exposure of college students to advocate reducing the intake of snacks.2.The results showed that the natural polyphenol LRP could protect PC12cells from oxidative stress induced by H₂O₂.LRP can be reduced by ROS level,protect the normal function of mitochondria and inhibit the activation of Caspase-3/-8/-9 to reduce the oxidative stress injury and apoptosis of cells.SH-SY5Y cells are induced by ACR to establish a neural injury model,the results show that LRP can improve the protein expression of GCLC,GCLM and HO-1,which are antioxidant genes downstream of Nrf2 pathway.