The Related Neuroprotective Mechanism Of Hydrogen-Rich Water On The Traumatic Brain Injury Rats

Part 1 The Related Neuroprotective Mechanism Of Hydrogen-Rich Water On The Traumatic Brain Injury RatsObjective:To determine the effect of hydrogen-rich water on inflammation and apoptosis in the penumbra tissue of brain injury of rats.Methods:a total of 54 adult male Sprague-Dawley(SD)rats were randomly divided into three groups:sham-operated,TBI(Traumatic brain injury),and TBI+HW(Traumatic brain injury+ hydrogen-rich water).Each group is divided into 24h,3d and 7d three subgroups.Sham group Don’t impact.After inducing TBI,TBI+HW group intraperitoneal inject hydrogen-rich water(5 ml/kg),and one times a day until to die.Sham group and TBI group at the same time point intraperitoneal inject same amount of saline solution.Observe neurological severity scores(NSS)for neurologic deficits,hematoxylin-eosin staining for pathological changes,apoptotic cells and apoptotic index detected by TUNEL,and the expression of inflammatory factors(TNF-a、IL-β)in penumbra tissue were detected by enzyme linked immunosorbent assay(ELISA).Results:The NSS scores of TBI group and TBI+HW group were significantly higher than sham group.3d and 7d after injury,The NSS scores of TBI+HW group were significantly lower than TBI group.in TBI+HW group,The necrosis area of brain tissue was reduced and the brain edema was alleviated.Compared with TBI group,the apoptosis index and the expression of inflammatory factors(TNF-a、IL-1β)in TBI+HW group were significantly decreased.CONCLUSION:Hydrogen-rich water can inhibit the expression of inflammatory factors and the occurrence of apoptosis in the penumbra tissue of brain after TBI,Hydrogen-rich water can alleviate the injury of nerve cells and improve the nerve function after TBI.Part 2 The Effect of hydrogen-rich water on inflammatory response mediated by TLR4/MyD88/NF-KB signaling pathway in the penumbra tissue of brain injury of rats.Objective::To observe the effect of hydrogen-rich water on the inflammatory response mediated by TLR4/MyD88/NF-KB signaling pathway in the penumbra of rats with traumatic brain injury.Methods:a total of 54 adult male Sprague-Dawley(SD)rats were randomly divided into three groups:sham-operated,TBI(Traumatic brain injury),and TBI+HW(Traumatic brain injury+hydrogen-rich water).Each group is divided into 1d,3d and 7d three subgroups.Sham group Don’t impact.After inducing TBI,TBI+HW group intraperitoneal inject hydrogen-rich water(5 ml/kg),and one times a day until to die.Sham group and TBI group at the same time point intraperitoneal inject same amount of saline solution.Observe The protein,mRNA expression of TLR4,MyD88,NF-KB and inflammatory factors(TNF-a、IL-1β)in the penumbra of brain.the expression of TLR4 positive cells in the penumbra were also observed.Results:Compared with sham group,The expression of TLR4,MyD88,NF-KB,inflammatory factor concentration and TLR4 positive cell rate in TBI group and TBI+HW group were significantly increased.Compared with TBI group,The expression of TLR4,MyD88,NF-KB protein,mRNA,inflammatory factor concentration and TLR4 positive cell rate in TBI+HW group were significantly decreased.CONCLUSION:Hydrogen-rich water can regulate the TLR4/MyD88/NF-KB signaling pathway of nerve cells of TBI,and then regulate the acute neuroinflammation of TBI to play a neuroprotective role.The specific mechanism may be through the regulation of TLR4/MyD88/NF-KB signaling pathway in neurons,especially glial cells.Part 3 Effect of hydrogen-rich water on mitochondrial apoptosis in penumbra of rats with traumatic brain injuryObjective:To observe the Effects of hydrogen rich water on mitochondrial pathway apoptosis in the penumbra of rats with traumatic brain injury.Methods:a total of 54 adult male Sprague-Dawley(SD)rats were randomly divided into three groups:sham-operated,TBI(Traumatic brain injury),and TBI+HW(Traumatic brain injury+hydrogen-rich water).Each group is divided into 1d,3d and 7d three subgroups.Sham group Don’t impact.After inducing TBI,TBI+HW group intraperitoneal inject hydrogen-rich water(5 ml/kg),and one times a day until to die.Sham group and TBI group at the same time point intraperitoneal inject same amount of saline solution.Observe The expression of reactive oxygen species(ROS),mitochondrial membrane potential(MMP),mitochondrial membrane permeability(MPTP)in each group.BAX and BCL-2 in penumbra of rats in each group were also observed.Results:Compared with sham group,the expressions of ROS,MMP and BAX in TBI+HW group and TBI+HW group were significantly increased,and MPTP,BCL-2 were also significantly increased.Compared with TBI group,the expression of ROS,MMP and BAX in TBI＋HW group were significantly decreased,and the expression of MPTP and BCL-2 were significantly increased.Conclusion:In the early stage of TBI,mitochondrial ROS increased,MMP decreased,mitochondrial permeability increased and the expression of apoptosis proteins Bax and Bcl-2 were abnormal,which were closely related to neuronal apoptosis,and these may be related to the positive regulation of NF-KB activation.hydrogen-rich water can alleviate the occurrence of mitochondrial pathway apoptosis mediated by NF-kappa B,thereby reducing neuronal apoptosis after TBI,thus achieving brain protection.