Anti-inflammatory Mechanism Of Mailuoning On Astrocytes And Neurons Of SD Rats Subjected To Oxygen Glucose Deprivation

Objective:To investigate the protective effects and possible anti-inflammatory mechanisms ofMailuoning on co-cultured neurons and astrocytes of SD subjected to Oxygen/Glucose Deprivation.Methods:Primary culture and identification of SD neurons and astrocytes were carried out,andthe co-cultured system of neurons and astrocytes was established by Transwell.The morphology of cells was observed by inverted microscope,and the neurons and astrocytes were identified by confocal microscope.To establish a model of co-cultured neurons and astrocytes of SD subjected to Oxygen/Glucose Deprivationby?an in vitro model of brain ischemia?using a physical method?95%N₂+5%CO₂?.CCK-8 and LDH methods was used to screen the optimal concentration of Mailuoning and to determine the protective effect of Mailuoning pretreatment with different concentrations on the neurons and astrocytes subjected to Oxygen/Glucose Deprivation.We use western blot to detect the protein expression of GLT-1 in the presence or absence of DHK.ELISA method was used to detect the expression of inflammatory factors TNF-?and IL-1?in the supernatant of co-cultured neurons and astrocytes after OGD/R.The protein expression of p-NF-?B p65 and I?B?was detected by western blot.Results:.High purity of neurons and astrocytes were successfully isolated and cultured,and theco-cultured system of neurons and astrocytes was established successfully;Pretreatment in the presence of Mailuoning can significantly increase the survival rate and decrease LDH leakage of neurons and astrocytes subjected to oxygen/glucose deprivation;The expression of glutamate transporter?GLT-1?protein in astrocytes was significantly increased after oxygen/glucose deprivation;The expression of inflammatory factor TNF-?,IL-1?was significantly reduced,indicating that Mailuoning has anti-inflammatory effect.The expression of inflammatory factor TNF-?and IL-1?were decresed after the glutamate transporter inhibitor?DHK?was added,the results indicating the protective effects of Mailuoning on the neurons and astrocytes associated with the glutamate transporter?GLT-1?.Conclusions:Mailuoning injection has protective effects on neurons and astrocytes induced byoxygen/glucose deprivation.The mechanism may be related to the increase of the expression of glutamate transporter protein,the decrease of the expression of inflammatory factors and the inhibition of NF-?B activation in astrocytes.