目的 探讨芹黄素对小胶质细胞活化的抑制作用.方法 原代培养SD大鼠小胶质细胞,实验随机分为空白对照组、脂多糖组(LPS组)、LPS+芹黄素(10 μM)组、LPS+芹黄素(20μM)组、LPS+芹黄素(50 μM)组.MTT法检测芹黄素对小胶质细胞活性的影响;ELISA法检测IL-1、IL-10等炎症相关因子以及BDNF、PDNF等神经营养因子的表达;RT-PCR、western blot检测炎症相关基因iNOS转录以及表达水平.结果 MTT检测结果表明,芹黄素对小胶质细胞活性无明显抑制.LPS刺激后,芹黄素预处理组IL-1等炎症因子表达水平明显低于单独LPS组(P<0.05);而芹黄素预处理组IL-10的表达则高于单独LPS组(P<0.01);与此同时,BDNF、PDNF等神经营养因子的分泌也有相同趋势(P<0.05).芹黄素预处理组iNOS表达和转录水平也低于单独LPS组(P<0.05).结论 芹黄素可抑制活化状态小胶质细胞炎症因子以及炎症相关蛋白的表达,并可同时促进与神经元分化成熟相关神经营养因子的分泌.
Objective To investigate the inhibitory effect of apigenin on the activation of microglial cells.Methods Microglial cells from Sprague-Dawley rats were cultured and randomly divided into blank control group,lipopolysaccharide (LPS) group,and LPS + apigenin (10 μM,20 μM,and 50 μM) groups.MTT assay was used to analyze the effect of apigenin on the viability of microglial cells;ELISA was used to measure the expression of inflammatory factors interleukin-1 (IL-1) and interleukin-10 (IL-10) and neurotrophic factors brain-derived neurotrophic factor (BDNF) and parasite-derived neurotrophic factor (PDNF);RT-PCR and Western blot were used to measure the transcriptional and expression levels of the inflammation-related gene inducible nitric oxide synthase (iNOS).Results MTF assay showed that apigenin had no marked inhibitory effect on the viability of microglial cells.After LPS stimulation,the apigenin pretreatment group had significantly lower expression of IL-1 and significantly higher expression of IL-10 than the LPS group (P 〈 0.05),and the secretion of BDNF and PDNF showed a similar trend (P 〈 0.05).The apigenin pretreatment group also had significantly lower transcriptional and expression levels of iNOS than the LPS group (P 〈 0.05).Conclusions Apigenin can inhibit the expression of inflammatory factors and proteins and promote the secretion of neurotrophic factors associated with neuron differentiation and maturation in microglial cells.