丁酸钠抑制氟中毒可诱导小胶质细胞活化及炎症因子表达增多

Inhibitory effects of sodium butyrate on microglial activation and expression of inflammatory factors induced by fluorosis

背景:在中枢神经系统疾病中,致病因素刺激小胶质细胞过度活化分泌炎性因子继而导致神经元进一步损伤,去乙酰化酶抑制剂丁酸钠能否降低氟中毒引起小胶质细胞的过度活化以及引起的炎性作用,值得深入研究。目的:丁酸钠对氟诱导的小胶质细胞过度活化以及炎性因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α表达的影响。方法:用含体积分数为10%胎牛血清的DMEM/F12培养基常规培养BV-2小胶质细胞,取对数生长期细胞用CCK-8法观察不同浓度氟化钠、丁酸钠及二者联合使用对BV-2细胞活力的影响。通过倒置显微镜、RT-qPCR和Western blot法观察氟化钠、丁酸钠及二者联合应用后的BV-2细胞形态以及炎症因子白细胞介素1β、白细胞介素6、肿瘤坏死因子αmRNA和蛋白表达。结果与结论:①CCK-8结果:质量分数25×10^-6氟化钠能降低BV-2细胞活力,0.25 mmol/L丁酸钠能升高BV-2细胞活力;0.25 mmol/L丁酸钠能逆转25×10^-6氟化钠对BV-2细胞活力的降低作用;②RT-qPCR和western blot结果:0.25 mmol/L丁酸钠能够拮抗25×10^-6氟化钠引起的白细胞介素1β、白细胞介素6、肿瘤坏死因子α过表达作用;③结果说明在一定的剂量范围内,丁酸钠可减轻氟致小胶质细胞介导的炎症反应。

BACKGROUND:In the diseases process of the central nervous system,pathogenic factors stimulate microglia cells to over-activate and secrete inflammatory factors,which lead to further injury of neurons.Whether the deacetylase inhibitor sodium butyrate can reduce the over-activation and inflammatory effects of microglia cells caused by fluorosis is worthy of further study.OBJECTIVE:To investigate the effects of sodium butyrate on fluorine-induced over-activation of microglia in the brain and the expression of inflammatory factors interleukin-1β,interleukin-6 and tumor necrosis factor-α.METHODS:Microglia cells(BV-2)were routinely cultured in F12/DMEM medium containing 10%fetal bovine serum.CCK-8 method was used to observe the effects of sodium fluoride,sodium butyrate and their combination on the viability of BV-2 cells in logarithmic growth phase.The morphology of BV-2 cells after treatment with sodium fluoride,sodium butyrate and their combination,mRNA and protein expressions of interleukin-1β,interleukin-6 and tumor necrosis factor-αwere observed by inverted microscope,RT-qPCR and western blot assay.RESULTS AND CONCLUSION:(1)CCK8 results showed that 25×10^-6 sodium fluoride could reduce the viability of BV-2 cells,and 0.25 mmol/L sodium butyrate could increase the viability of BV-2 cells.Sodium butyrate 0.25 mmol/L could reverse the decreased viability of 25×10^-6 sodium fluoride on BV-2 cells.(2)RTqPCR and western blot assay results showed that 0.25 mmol/L sodium butyrate could antagonize the over-expression of interleukin-1β,interleukin-6 and tumor necrosis factor-αinduced by 25×10^-6 sodium fluoride.(3)It indicates that sodium butyrate,in a certain dose,can reduce the inflammatory response mediated by fluorium-induced microglia cells.