Effects Of Incretin Receptor Agonists On Oxidative Stress In Cerebral Ischemia-reperfusion Model Rats Via Nrf2/ARE Pathway

Objective:1.To investigate the neuroprotective effects of GLP-1/GIP double receptor agonist DA3-CH and GLP-1 single receptor agonist Liraglutide on cerebral ischemia-reperfusion model rats(neurological deficit score,infarct size).2.To explore the effects of DA3-CH and Liraglutide on oxidative stress in cerebral ischemia-reperfusion model rats,and to explore the relationship between its mechanism and Nrf2/ARE pathway(Nrf2-related pathway proteins and oxidative stress related proteins).3.To explore and compare the neuroprotective effect of DA3-CH and Liraglutide on cerebral ischemia-reperfusion model rats and the effect on oxidative stress.Methods:1.Grouping: 72 male SD rats were randomly divided into 4 groups:group A(Sham group,n=18),group B(Model group,n=18),group C(Liraglutide group,n=18)and group D(DA3-CH group,n=18).2.Intervention methods: Under the same feeding environment,group C and D were intraperitoneally injected with Liraglutide(10nmol/kg,once A day)and DA3-CH(10nmol/kg,once A day)for 14 consecutive days,and groups A and B were intraperitoneally injected with equal volume of normal saline.3.Assay methods: Blood glucose of rats in each group was measured after intraperitoneal injection for 14 consecutive days.Then group A was exposed only artery without embolization,and group B,C and D were prepared by cerebral ischemia reperfusion model(ischemia for 2 hours caused by middle artery embolization and reperfusion for 22 hours).The cerebral infarction area was determined by TTC staining.SOD activity and MDA content were measured by Elisa.Nrf2/ARE pathway related proteins were measured by Western-blot method.Results:1.Blood glucose level: there was no significant change in blood glucose among all groups(P>0.05).2.Neurological deficit scores: Compared with group A(Sham group),the neurological deficit scores of the other three groups were significantly higher(P<0.001),and the neurological deficit scores of group C(Liraglutidez group)and group D(DA3-CH group)were lower than group B(Model group)(Liraglutide group,P<0.01;DA3-CH,P < 0.001).The neurological deficit score of group D was lower than that of group C(P<0.05).3.Cerebral infarction area size: compared with group A,the other three groups had obvious infarct focal tissue(P<0.001),and the percentage of cerebral infarction area in group C and D was smaller than that in group B(P<0.001).The percentage of cerebral infarction area in group D was smaller than that in group C(P<0.05).4.Oxidative stress related indexes in penumbra tissue: MDA content: Compared with group A,MDA content in the other three groups was significantly increased(P<0.001),and MDA content in groups C and D was significantly decreased compared with group B(P<0.001).MDA content in group D was lower than that in group C(P<0.01).SOD activity: Compared with group A,SOD activity in the other three groups decreased significantly(P<0.001),and SOD activity in groups C and D was higher than that in group B(Liraglutide,P<0.01;DA3-CH,P<0.001).SOD activity in group D was higher than that in group C(P<0.01).5.Nrf2-related pathway protein: Compared with group A,the expression of Nrf2 in ischemic penumbra tissue of the other three groups was significantly increased(P<0.001),and the expression of Nrf2 in group C and D was higher than that in group B(P<0.001).The expression of Nrf2 in group D was higher than that in group C(P<0.001).Similarly,compared with group A,the expressions of HO-1 and NQO-1 in ischemic penumbra tissue of the other three groups were significantly increased(P<0.001).The expressions of HO-1 and NQO-1 in group C and D were higher than those in group B(P<0.001).The expressions of HO-1 and NQO-1 in group D were higher than those in group C(P<0.001).Conclusion:1.Both DA3-CH and Liraglutide can reduce the infarct size and the neurological deficit score of cerebral ischemia-reperfusion model rats,and have a neuroprotective effect.2.Both DA3-CH and Liraglutide can reduce the oxidative stress response of cerebral ischemia-reperfusion model rats,and this effect is related to the activation of Nrf2/ARE pathway.3.The neuroprotective effect,inhibitory effect on oxidative stress response and activation of Nrf2/ARE pathway of DA3-CH were stronger than that of Liraglutide in cerebral ischemia-reperfusion model rats.