Study On The Protective Effect Of Resveratrol On H₂O₂-induced SH-SY5Y Cells Damage Based On Energy Metabolism

As the aging of the population accelerates,the incidence of neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease is increasing year by year,which seriously threatens the health of the elderly and also brings a heavy burden to society and families.The energy supply of neuronal cells have shown that closely related to their functions.Insufficient energy supply will affect nerve function and lead to neurodegenerative diseases.Resveratrol is a ubiquitous polyphenolic compound.A lot of studies have shown,it can protect nerve,anti-oxidative damage and anti-inflammatory activities.However,most studies mainly focus on antioxidant activity.Whether the resveratrol protecting nerve effects related to the influence of intracellular energy metabolism,the specific mechanism is unclear.The purpose of this experiment is to study the protective effects of resveratrol on H2O2-induced oxidative damage to human neuroblastoma cells(SH-SY5Y)cell model and its relationship with energy metabolism.It provides new ideas for the research on the neuroprotective mechanism of resveratrol,and has certain guiding significance for the development of drugs for neurodegenerative diseases.The experiment were selected to study the protective effect of resveratrol on the H2O2induced oxidative damage model of SH-SY5Y nerve cells and its possible mechanisms related to energy metabolism at the level of energy metabolism.The main experimental methods are as follows:1.Treated SH-SY5Y cells with 1.2 mmol/L H2O2 for 24 h to establish an oxidative damage model;20,10,5 and 1 μmol/L resveratrol were pretreated for 24 h,added 1.2 mmol/L H2O2 for oxidative damage.The role of resveratrol was explored measured by MTT and growth curves;2.PI single staining method detecting the effects of resveratrol on the cells cycle oxidative damage cell models;3.Annexin V single staining method detecting the effects of resveratrol on the apoptosis rates of H2O2 oxidative damage models;4.HE staining to observe the effects of resveratrol on the morphological changes of oxidative damage model cells;5.The enzyme activity detection kit to detect the effects of resveratrol on the activitivies of SOD,GSH-Px and the content of MDA in the H2O2 oxidative damage cell model;6.To detect the effects of resveratrol on the energy metabolism related enzymes HK,PFK,PK,SDH and LDH activities,energy metabolite ATP content and glucose consumption.7.Western blot detect of the effects of resveratrol on the expression of HIF-lα,MCT-1 and GLUT-1 in H2O2 oxidative damage cell models.The experimental results are as follows:1.The cell viability of SH-SY5Y cells decreased in H2O2 injury group by the MTT and growth curves.Cell viability increased in the resveratrol group(P<0.01);2.The 20,10 and 5μmol/L resveratrol pretreatment group compared with the oxidative damage model group,G0/G1 phase and G2/M phase were significantly increased(P<0.01),and the percentage of S phase was significantly reduced(P<0.01);3.The results of HE staining showed that the morphological changes of cells pretreated with 20,10,and 5 μmol/L resveratrol were not obvious,which was similar to the control group.4.Compared with the H2O2 oxidative damage cell model group,the apoptosis rate of 20,10 and 5 μmol/L resveratrol was significantly reduced(P<0.01);5.In the SH-SY5Y cells,the SOD and GSH-Px activities pretreated with 20,10 and 5 μmol/L resveratrol increased significantly(P<0.01),and the MDA content was significantly decrease(P<0.01));6.The PFK,PK,SDH activities,glucose consumption and ATP content in the H2O2 oxidative damage cell model were significantly decreased(P<0.01),and the HK and LDH activities were significantly increased(P<0.01);Compared with the H2O2 oxidative damage group,the PFK,PK,SDH activity,ATP content and glucose consumption in the cells of the 20,10 and 5 μmol/L resveratrol pretreatment groups increased(P<0.01),and the HK and LDH activities were significantly higher decrease(P<0.01);7.Compared with the control group,the expression of HIF-la in the cells of the H2O2 oxidative damage group was significantly increased(P<0.01),and the protein expression of GLUT-1 and MCT-1 was significantly decreased(P<0.01);Compared with the H2O2 oxidative damage model group,cells pretreated with different concentrations expression of GLUT-1 and MCT-1 of resveratrol increased,and the protein content of HIF-la was significantly decreased(P<0.01).Above all experimental results,1.2 mmoL/L H2O2 can inhibit the proliferation of SH-SY5Y cells,change the cell cycles,and promote cells apoptosis.Pretreatment of cells with resveratrol at concentrations of 20,10 and 5 μmol/L can inhibit H2O2-induced cell apoptosis and cycles arrest.The enzymatic activity of PFK,PK,SDH in cells was increased,the content of energy metabolites ATP and glucose consumption.After resveratrol pretreatment,it reduces the effect of H2O2 on the expression of GLUT-1 and MCT-1 protein,and reduces HIF-1α protein expression level.In summary,H2O2 can inhibit the glucose uptake function of cells,reduce the activity of enzymes related to glucose metabolism,and affect the expression of key proteins to affect the energy metabolism process in nerve cells.And different concentrations of resveratrol reduce the effect of H2O2,regulates the activities of key glycolysis enzymes HK,PFK and PK.Improve the activity of SDH,an important enzyme of the tricarboxylic acid cycle.Increase cells productivity.Improve cells productivity and protect cells from oxidative damage induced by H2O2 by affecting the process of energy metabolism in the cells.