Dl-3-n-butylphthalide Regulates The Inflammatory Response Of BV2 Microglial Cells By Inducing Autophagy

Objective:To investigate the effect of dl-3-n-Butylphthalide on the inflammatory response of BV2 microglial cells induced by Aβ_25-355-35 and its possible mechanism.Methods:BV2 microglial cells were cultured and divided into blank control group（Control）,model group（Model）,dl-3-n-Butylphthalide group（NBP）and dl-3-n-Butylphthalide combined with autophagy inhibitor 6-amino-3-methylpurine group（3-MA）.CCK-8 assay was used to detect the effect of dl-3-n-Butylphthalide and Aβ_25-355-35 on the activity of BV2 microglial cells,Western blot was used to detect the protein expression levels of LC3B-Ⅱand p62.Real-time PCR was used to detect the expression levels of autophagy indicators ATG5、Beclin1 and inflammatory indicators IL-1β、IL-6.Results:The Aβ_25-355-35 concentration of 20?mol/L or more has significant damage to the cell survival rate,No significant difference of cell viability was found when the concentration of dl-3-n-Butylphthalide was no more than 40?mol/L.The mRNA expression levels of IL-1βand IL-6 in the Model group were significantly up-regulated compared with the Control group（P<0.05）.Compared with the Model group,The mRNA expression levels of IL-6 and IL-1βin the NBP group were significantly reduced,The mRNA expression levels of ATG5、Beclin1 and protein expression level of LC3B-Ⅱincreased significantly,the protein expression level of p62 decreased（P<0.05）,Compared with the NBP group,The mRNA expression levels of IL-6 and IL-1βin the 3-MA group were significantly increased,the mRNA expression levels of ATG5、Beclin1 and the protein expression level of LC3B-Ⅱdramatically decreased,the protein expression level of p62 increased（P<0.05）.Conclusion:Related pro-inflammatory factors were significantly expressed in BV2microglia treated with Aβ_25-35,indicating that Aβ_25-355-35 was the induced factor of microglia activation.dl-3-n-Butylphthalide can reduce the inflammatory response level of BV2 microglia induced by Aβ_25-35,and can also up-regulate the expression levels of autophagy related genes and proteins,the expression levels of inflammatory factors were significantly increased when the autophagy effect was inhibited.This suggests that dl-3-n-Butylphthalide may play a neuroprotective role by inducing autophagy to down-regulate BV2 microglia inflammatory response level.