Resveratrol Inhibits Autophagy And Improves Chronic Cerebral Ischemic Cognitive Dysfunction

Chronic cerebral ischemia(CCH)is a common clinical cerebral ischemia injury,which is a pathological factor in the occurrence and development of vascular dementia,Alzheimer's disease,Binswanger's disease and other neurological diseases or mental diseases.According to reports,long-term cerebral blood perfusion deficiency leads to energy metabolism disorder,oxidative stress production,inflammatory factor release,neuronal apoptosis,and then causes cognitive impairment.The results show that,after chronic cerebral ischemia,inhibition of oxidative stress,promotion of angiogenesis and establishment of effective collateral circulation are effective methods to improve the global and local cerebral hypoperfusion and reduce the damage of nerve function.Resveratrol is a kind of plant antitoxin,which belongs to natural polyphenols.It has a series of protective effects,such as anti-inflammatory,anti-oxidation,anti-diabetes,myocardial protection and so on.It has been proved that resveratrol can protect the brain nerves from oxidative damage by reducing oxidative stress and improve the cognitive function of mice with Alzheimer's disease,but whether it can improve the cognitive function of chronic cerebral ischemia is still unclear.The aim of this study is to establish a CCH rat model and to explore the effect of resveratrol on the cognitive function of CCH rats.1.Resveratrol improves chronic cerebral ischemic cognitive dysfunctionObjective: To explore the protective effects of resveratrol on chronic cerebral ischemic cognitive dysfunction.Methods: SD rats were randomly divided into sham operation group(Sham group),chronic cerebral hypoperfusion model group(CCH group),resveratrol + chronic cerebral hypoperfusion group(Res group);2CH method was used in CCH group and Res group.A rat model of chronic cerebral ischemic cognitive dysfunction was established.The Res group was orally administered with resveratrol 50 mg/kg daily after modeling.Bederson neurological deficit score and Morris water maze were used at 3,6,and 9 weeks,respectively.The learning and memory abilities of rats in each group,HE evaluation of histopathological damage,ELISA detection of brain injury markers and changes of oxidative stress factors,TUNEL detection of neuronal apoptosis,immunofluorescence detection of MAP2 expression,observation of neuron survival,microvascular Density and angiogenesis.Results: At 3,6,and 9 weeks after the onset of ischemia,the Bederson score of the rats in the CCH group was significantly increased,the incubation period was prolonged,and the swimming distance was increased.Compared with the Sham group,the differences were significant(P <0.05),indicating that the learning and memory abilities of the rats were decreased.After resveratrol intervention,the cognitive function was significantly improved,the cell structure and neuron morphology of the frontal cortex and hippocampus of rats were repaired,the neuronal degeneration and necrosis phenomenon was improved,the brain injury markers were reduced,and oxidative stress injury was suppressed;In addition,the expression of MAP2 in the Res group was significantly increased,and the apoptosis of neurons in the hippocampus was reduced.Conclusion: Resveratrol can reduce brain damage,inhibit oxidative stress injury,reduce neuronal cell apoptosis,and improve learning and memory ability in CCH rats.2.Resveratrol inhibits autophagy through PI3K/AKT/mTOR pathway to improve chronic cerebral ischemic cognitive dysfunctionObjective: To explore the effect of resveratrol on autophagy in CCH rats,and to explore the regulatory mechanism of PI3K/AKT/mTOR.Methods: Immunofluorescence and Western blot were used to detect apoptosis-related proteins Bcl-2,Bax and cleaved Caspase3,expressions of autophagy-related proteins Beclin1,LC3II/I,and PI3K/AKT/mTOR pathway-related proteins.On this basis,a resveratrol + chronic ischemia + PI3 K inhibitor LY294002 group(PI3K group)was established.Morris water maze was used to detect the learning and memory abilities of rats in each group.HE staining was used to observe brain tissue damage and ELISA was used to detect oxidative stress injury.Western blot was used to detect apoptosis and autophagy related proteins.Results: The expressions of pro-apoptotic factors Bax and cleaved Caspase3 in the Res group were significantly reduced,and the expression of anti-apoptotic factors Bcl-2 was increased(P <0.05).The results of autophagy-related protein detection showed that the autophagy factor expression in the brain group of the CCH group was significant.The expression of Beclin1 and LC3II/I was significantly reduced after resveratrol treatment(Vs.CCH,P <0.05);meanwhile,resveratrol activated PI3K/AKT/mTOR in the signal pathway,PI3 K expression was significantly increased,and AKT and mTOR phosphorylation levels were significantly increased(Vs.CCH,P <0.05).PI3 K inhibitor LY294002 was given to CCH rats together with resveratrol.As a result,rats in the PI3 K group recognized The cognitive and spatial memory ability is poor;the brain tissue of the rat's frontal cortex and hippocampus relaxes,and neuronal damage is most obvious;oxidative stress injury is not alleviated,the apoptosis rate increases,and the expression of pro-apoptotic factors increases Excessive autophagy has not been suppressed.Conclusion: Resveratrol can inhibit the autophagy level of CCH rats by improving the PI3K/AKT/mTOR signaling pathway and improve the learning and memory ability of CCH rats.3.Resveratrol activates Slit2/Robo4 pathway to improve angiogenesis and relieve chronic cerebral ischemic cognitive dysfunctionObjective: To explore the effect of resveratrol on angiogenesis in CCH rats,and to explore the mechanism of Slit / Robo pathway regulation.Methods: Western blot and q RT-PCR were used to detect the expressions of Slit2,Robo4 related proteins Slit2,Robo4 and vascular endothelial related factors ET-1,VEGF,SDF-1? and e NOS;establish resveratrol + chronic cerebral ischemia + Robo resistance Decoction(SR group),immunofluorescence detection of MAP2 expression to evaluate vascular injury and repair;assessment of neural function score,water maze test to detect cognitive function.Results: The expression of Slit2 / Robo4 related proteins of Slit2 / Robo4 pathway was detected by PCR and Western blot.The results showed that compared with the Sham group,the expression of Slit2 and Robo4 in the CCH group was increased.Compared with the CCH group,the expression of Slit2 and Robo4 in the Res group was further increased.Increased(P <0.05);After the Robo blocker was administered,MAP2 expression decreased in the SR group.Compared with the CCH group,the expression of e NOS,VEGF,and SDF-1? in the Res group was increased,and the expression of ET-1 was decreased.There was a statistical difference between the groups(P <0.05).After adding the Robo inhibitor,Res related factors at the same time,Robo inhibitors reversed the effect of resveratrol on neurological score and cognitive level.Conclusion: Resveratrol can activate the Slit2 / Robo4 pathway,improve vascular endothelial function,and improve cognitive dysfunction in CCH rats.