法舒地尔通过调控糖酵解通路对原代小胶质细胞的影响

Effects of Fasudil on Primary Microglia by Modulating the Glycolytic Pathway

目的探讨法舒地尔在阿尔茨海默病(Alzheimer disease,AD)等多种认知功能障碍的疾病中对原代小胶质细胞的作用及其可能的调控机制。方法采用原代小胶质细胞,在经过不同浓度的法舒地尔处理后,荧光定量PCR检测细胞炎症水平、糖酵解相关酶及HIF-1α的活化水平,酶标仪检测原代小胶质细胞Aβ吞噬活化能力,通过ATP实验检测小胶质细胞的能量代谢能力。结果经过不同浓度的法舒地尔处理后,LPS刺激不同时间点的原代小胶质细胞的促炎因子的mRNA表达水平均有下降(P<0.05);Aβ吞噬能力明显增加(P<0.001);法舒地尔促使原代小胶细胞细胞新陈代谢能力明显增加(P<0.005),糖酵解过程的相关酶以及HIF-1α的mRNA水平均有下降(P<0.005)。结论法舒地尔可能通过下调小胶质细胞HIF-1α的转录活性来降低糖酵解过程相关酶的相对表达量,抑制糖酵解的过程从而增加ATP的产生,维持小胶质细胞的新陈代谢来减轻炎症反应,增加吞噬作用。

Objective To investigate the role of fasudil on primary microglia and its possible regulatory mechanisms in a variety of cognitive disorders such as Alzheimer disease(AD).Methods In this study,microglia cells were treated with different concentrations of fasudil,fluorescence quantitative PCR was used to detect cellular inflammation levels and activation levels of glycolysis-related en⁃zymes and HIF-1α,enzymatic marker was used to detect Aβphagocytic activation of microglia cells,and energy metabolism of microg⁃lia cells was detected by ATP assay.Results After treatment with different concentrations of fasudil,the mRNA expression levels of proinflammatory factors were decreased(P<0.05);Aβphagocytic capacity was significantly increased(P<0.001);the metabolic capaci⁃ty of microglia cells was significantly increased(P<0.005);the mRNA levels of involved in glycolysis and HIF-1αwere decreased(P<0.005).Conclusion Fasudil may reduce the relative expression of enzymes associated with the glycolytic process by down-regulating the transcriptional activity of HIF-1αin microglia,inhibit the glycolytic process and thereby increase ATP production and maintain mi⁃croglia cell metabolism to reduce inflammatory responses and increase phagocytosis.