Effect Of Nesfatin-1 On ExPression Of CAMK? And PSD95 Protein After Cerebral Ischemia-Reperfusion In Rats

Objective: To investigate the effect of Nesfatin-1 on the ex Pression of camkii and Psd95 Protein after ischemic rePerfusion in middle cerebral arteries of rats(middle cerebral artery occlusion rePerfusion mcao /r),to investigate whether Nesfatin-1 can Protect the ischemic rePerfusion Part of rat brain,and whether it may cause the difference of camkii and Psd95 Protein ex Pression.Methods: Results: healthy male sd rats were randomly divided into normal control grou P(n =8),model grouP: mcao/r grouP(n =8),intervention grouP: mcao/rNesfatin-1 1?g/100g(n =8),mcao/r Nesfatin-1 5?g/100 g grouP(n =8),and other 8 s Pares.In the model grouP,10% chloral hydrate was injected into the middle cerebral artery for 24 hours and the rats were treated with modified embolization method.When the rats were in the best state of anesthesia,that is,the rats were fixed to the fixed Plate after the reversal of Positive reflex,the rats were treated with local dissection in the neck.The normal control grouP isolated only the right common carotid artery,external carotid artery,internal carotid artery,isolated vagus nerve,debridement and suture skin.after the o Peration was comPleted,the rat brain tissue was removed after 24 hours of re Perfusion,ttc staining was Performed,the area of cerebral infarction was calculated by PhotograPhing,he and immunohistochemical staining were Performed to observe the morPhological changes of neuronal cells and the Positive ex Pression of camkii and PSD95 in the tissue cells of immunohistochemical sections.the light density value of ihc sections was calculated.the Protein extracted from fresh brain tissue of rats was taken.the Protein ex Pression of camkiii,PSD95 was observed by wb Protein imPrinting method.the gray value of Protein band was analyzed by image-j and graPhPad software.Result: 1.The results of HE staining under light microscoPe showed that comPared with the normal control grouP,the MCAO/R model grouP showed changes of brain tissue with stress,loose edema of brain tissue,microvascular and ca Pillary dilatation,congestion,inflammatory cell infiltration and nerve fiber swelling.com Pared with the normal control grouP,the edema condition was slightly im Proved in the mcao/rNesfatin-11?g/100 g grouP,but the edema brain tissue was still clearly visible.it was difficult to observe edema neurons and nerve fibers in the mcao/rNesfatin-15?g/100 g grouP,no tissue necrosis,disintegration and no infiltration of inflammatory cells.2.Immunohistochemical assays for CAMKII and PSD95 Protein exPression showed that comPared with the normal control grouP,the model grouP CAMKII and PSD95 showed weakly Positive ex Pression in the cell membrane after ischemia-rePerfusion,comPared with the Nesfatin-1 intervention grouP and the model grouP,CAMKII and PSD95 showed Positive ex Pression in the cell membrane but lower than the normal control grou P(P<0.05).camkii and PSD95 are mainly distributed in the cell membrane,where camkii is mainly distributed in the PresynaPtic cell membrane,and PSD95 is mainly distributed in the PostsynaPtic cell membrane.at the time of ischemic change,the ex Pression of camkii and PSD95 is inhibited,and the Positive ex Pression of cell membrane is not obvious.3.The results of TTC staining showed that Nesfatin-1 could reduce the infarct area of rat brain tissue,reduce the edema of brain tissue,and the normal control grou P had no changes of cerebral infarction edema,and the right ischemia re Perfusion site of MCAO/R model grouP showed a large area of white brain tissue staining,which indicated that the right cerebral tissue of model grouP had infarction and stress injury caused by rePerfusion.4.Western-Blot test results showed that comPared with the normal control grouP,the exPression of CAMKII and PSD95 Protein showed a down-regulation trend(P<0.05),the MCAO/RNesfatin-11?g /100 g grouP comPared with the model grouP,the exPression of MCAO/RNesfatin-15 g/100 g grouP showed a certain uP-regulation trend,(P <0.05).Conclusions: 1.According to the results of HE staining,cerebral ischemia-rePerfusion induced ischemic infarction and stress edema of nerve cells and nerve fibers caused by rePerfusion.Nesfatin-1 could imProve the neuroedema status of neurons in ischemiarePerfusion and decrease the damage with the increase of dosage.2.ComPared with the blank control grouP,the Protein exPression of CAMKII and PSD95 in the hiPPocamPus and cortex of the rats in the model grouP was weakly Positive,and the Positive degree of intervention histone was lower than that in the blank control grou P but higher than that in the model grouP.3,camkii and PSD95 are very sensitive to ischemia,and their ex Pression will decrease after ischemia.Nesfatin-1 can effectively alleviate the a PoPtosis,necrosis,disintegration and edema of nerve cells in rat brain tissue after ischemia and rePerfusion,and then effectively enhance the Protein ex Pression level of camkii and PSD95,which may be related to the ability of Nesfatin-1 to reduce oxidative stress injury.Nesfatin-1,as a drug used only in the study of renal ischemia-rePerfusion injury and myocardial ischemiarePerfusion injury,has a certain reference value for the study of the mechanism of cerebral ischemia-rePerfusion injury and the later develo Pment of clinical treatment drugs.