Study Of Rho Kinase Isoforms In Enthanol Induced Brain Cellular Damage By Change Of AQP4 And AQP9 Expression

Objective: Ethanol is the most common poison and one of the important factors involved in and affecting brain injury.Long-term heavy drinking can cause changes in brain structure,degeneration of neurons in the brain,and affect quality of life.There is currently no effective treatment for alcoholic brain injury.Therefore,studying the pathogenesis of alcohol-induced brain cell injury has important clinical significance for the prevention and treatment of alcoholic encephalopathy.Ethanol can cause brain cell damage through multiple links,but its specific action is very complicated.In addition to the direct neurotoxic effects of ethanol,its indirect damage is secondary brain cell injury.AQP4 But the mechanism of how alcohol causes brain cell injury is not clear.Methods:1.Establishment of rat model of chronic alcoholic brain injury.2.Primary culture of glial cells.3.Lentivirus was used to interfere with ROCK isoforms.Real-time PCR and Western blot were used to detect the expression of ROCK1,ROCK2,NF-?B p65,AQP4,and AQP9 in the brain tissue of rats in each group;meanwhile,ROCK1 in brain glial cells was detected in vitro,ROCK2,NF-?B p65,AQP4,and AQP9 expression.4.Cells and animal models were treated with NF-?B inhibitor PDTC.With the help of two different methods,PCR and blot Western,the gene expression levels of AQP4,AQP9 and-NF?Bp65 three different protein molecules in the rat brain tissue of each experimental group were determined and analyzed..At the same time,NF-?B p65,AQP4 and AQP9 expression in brain glial cells in vitro was detected.Result:1.The rat model of chronic alcoholic brain injury was successfully established;the primary culture of glial cells was successfully completed.2.Alcohol significantly increased the content of two different protein molecules,-TNF?and 6-IL,in the brain tissue of experimental mice,while the content of 10-IL decreased significantly;after treatment with lentivirus interference ROCK isomers and NF-?B inhibitors,TNF-? and IL-6 content can be significantly reduced.3.After the action of ethanol,the expression of ROCK isoforms and NF-?B increased,and,AQP4 and AQP9 reduced,and brain cells became injury.After the lentivirus interfered with ROCK isoforms,the expression of ROCK1 and ROCK2 was suppressed,The gene expression of the NF-?B protein molecule is significantly down-regulated,while the gene expression of the two protein molecules of AQP4andAQP9 is significantly up-regulated,and the damage of brain tissue cells is reduced to a certain extent.;AQP4 and AQP9 increased after treatment with NF-?B inhibitors,and brain cells injury could be significantly reduced;Conclusions:1.ROCK isomers are involved in the regulation of ethanol-induced brain cells injury.2.NF-?B participates in the process of ethanol-induced brain tissue damage by regulating the expression of AQP4 and AQP9,and it is a downstream signaling molecule of Rho kinase.