DHA对七氟醚诱导小胶质细胞活化的影响

Effect of DHA on sevoflurane-induced activation of microglia

目的评价DHA对七氟醚诱导小胶质细胞活化的影响.方法N9小鼠小胶质细胞,以1×106/ml的密度接种于培养板(1ml)或培养皿(10 ml)中,采用随机数字表法分为4组(n=18):对照组(C组)、DHA组、七氟醚组(Sevo组)和DHA+七氟醚组(DHA+Sevo组).C组不给予药物处理;DHA组和DHA+Sevo组在含25μmol/L DHA培养基中培养;SeVo组和DHA+Sevo组置于2%七氟醚处理装置中培养.于培养24 h时采用免疫组化法检测活化小胶质细胞并计算CD11b阳性细胞率;Western blot法检测活化小胶质细胞标志物CD11b的表达;ELISA法测定TNF-α和IL-1β的浓度.结果与C组和DHA组比较,Sevo组CD11b阳性细胞率增加,CD11b表达上调,TNF-α和IL-1β浓度升高(P<0.05);与Sevo组比较,DHA+Sevo组CD11b阳性细胞率降低,CD11b表达下调,TNF-α和IL-1β浓度降低(P<0.05).结论DHA可通过减少七氟醚诱导的小胶质细胞活化,降低炎性反应.

Objective To evaluate the effect of DHA on sevoflurane-induced activation of microglia.Methods N9 mouse microglia were seeded in culture plates(1 ml)or culture dishes(10 ml)at a density of 1×10^6 cells/ml and divided into 4 groups(n=18 each)using a random number table method:control group(C group),DHA group,sevoflurane group(Sevo group)and DHA plus sevoflurane group(DHA+Sevo group).Group C received no treatment.Cells were incubated in the culture medium containing 25μmol/L DHA in DHA and DHA+Sevo groups.Cells were exposed to 2% sevoflurane in Sevo and DHA+Sevo groups.At 24 h of culture,activated microglia were detected and counted by immunohistochemistry,the rate of CD11b positive cells was calculated,the expression of microglial biomarker CD11b was detected by Western blot,and the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-1beta(IL-1β)were determined by enzyme-linked immunosorbent assay.Results Compared with C and DHA groups,the rate of CD11b positive cells was significantly increased,the expression of CD11b was up-regulated,and the concentrations of TNF-α and IL-1β were increased in Sevo group(P<0.05).Compared with Sevo group,the rate of CD11b positive cells was significantly decreased,the expression of CD11b was down-regulated,and the concentrations of TNF-α and IL-1β were decreased in DHA+Sevo group(P<0.05).Conclusion DHA can decrease inflammatory responses through reducing sevoflurane-induced activation of microglia.