Electroacupuncture Improve The Cognitive Impairment Of Rats With Chronic Cerebral Hypoperfusion By Suppressing The Oxidative Stress Response And Brain Iron Deposition

Objective By extecting the expression of nerve injury genes,such as inflammation,oxidative stress and apoptosis,and the expression of signaling pathway of iron death,to provide experimental basis and effective methods of acupuncture intervention in the treatment of cognitive impairment caused by chronic cerebral hypoperfusion.Methods Morris water maze was used to analyze the spatial learning and memory ability of experimental rats and to evaluate the brain injury of rats.Western blot analysis of NOX1,NOX2 and NOX4 gene expression in rats.Real-time quantitative PCR was used to detect changes and screen microRNA.The interaction between microRNA and NOX genes was analyzed by bioinformatics and double luciferase experiments.Apoptosis of brain cells was observed by TUNEL staining.ELISA method to detect the levels of Ang,NOX,ROS,SOD,MDA,CAT and IL-6 in arterial blood and the brain tissue;The expression of microRNA was detected by qRT-PCR,and the expressions of NOX4,cox-2,eNOS and iNOS genes were detected by qRT-PCR and Western blot.The expression of NOX4 and GFAP was observed by immunofluorescence.Iron deposition in the brain after electroacupuncture was observed by Prussian blue staining,expression of miR485-3p and miR29 was detected by qRT-PCR,and PTGS2,ASCL4,IREB2,GPX4 and FPN gene expression was detected by qRT-PCR and Western blot.Results1.Electroacupuncture therapy can effectively improve the cognitive function of model animals.2.The expressions of NOX1,NOX2 and NOX4 in the model rats increased significantly compared with the sham group,and NOX1,NOX2 and NOX4 gene expression decreased significantly after electroacupuncture.3.The expression levels of the 9 miRNAs were significantly changed after electroacupuncture compared with the model group.The expressons of mir96,mir501-3p,mir9-5p,miR9,mir27a downregulated.And the expressions of mir132,mir181c,miR137,mir195 were upregulation.4.The bioinformatics analysis found that miR137 interacted with NOX4 gene,which was confirmed by double luciferase experiment.5.After electroacupuncture treatment,apoptosis was reduced,miR137 expression was significantly up-regulated,and related indicators of NOX gene,inflammation and oxidative stress were significantly downregulated.6.Electroacupuncture treatment can effectively improve brain iron deposition.7.GPX4 and FPN expression significantly up-regulated and expression of ASCL4.Ireb2,PTGS2 was significantly downregulated by electroacupuncture.The expression of miR29 was significantly upregulated and miR485-3p expression was significantly downregulated after electroacupuncture.Conclusion1.Electroacupuncture treatment improve CCH rats’ spatial learning and memory ability.2.Reducing the expression of NOX4 gene through the regulation of miR-137 expression,thus reducing the production of reactive oxygen species,which may be one of the mechanisms of electroacupuncture in the treatment of cognitive impairment.3.Electroacupuncture treatment reduce brain injury,inhibit the inflammation caused by cerebral ischemia and promote the regeneration of neurons and function recovery,prevent the excessive gliosis of astrocytes and promote the structural repair and functional recovery of nerve tissues.By regulating the expression of NOX4 gene by targeting miR137,the generation of reactive oxygen species can be reduced,the expression of nitric oxide synthase can be reduced,the oxidative stress response caused by cerebral ischemia can be alleviated,and the brain damage can be alleviated.Electroacupuncture therapy can effectively relieve brain injury and cognitive impairment caused by cerebral ischemia.4.Electroacupuncture therapy can improve the iron deposition caused by chronic cerebral hypoperfusion by affecting the expression of ASCL4,Ireb2,PTGS2,FPN and GPX4.Electroacupuncture treatment may affect the expression of IREB2 and FPN1 through mir485-3p and mir29.