瓜子金皂苷己抑制脂多糖诱导的小胶质细胞激活  

Polygalasaponin F Inhibits Lipopolysaccharide-induced Activation of Microglial Cells

作　　者：石瑞丽[1] 乌林哈德 齐瑞芳[1] 陈晓东[1] 高丽君[1] 秦文斌[1] 

机构地区：[1]包头医学院,内蒙古包头014040

出　　处：《中药新药与临床药理》2017年第3期263-267,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(No 81450058;30973887);内蒙古自治区自然科学基金项目(No 2013MS1108)

摘　　要：目的观察瓜子金皂苷己(polygalasaponin F,PGSF)对脂多糖(LPS)诱导的BV-2小胶质瘤细胞激活的作用及其作用机制。方法以0.1μg·mL^(-1)LPS刺激BV-2细胞24 h构建神经炎症模型,以酶联免疫吸附分析法(ELISA)检测细胞培养基上清中的白介素1β(IL^(-1)β)、肿瘤坏死因子(TNF-α)浓度,以Griess法检测一氧化氮(NO)的浓度;以逆转录-聚合酶链式反应(RT-PCR)法或免疫印迹法(Western Blot)检测细胞炎性蛋白酶诱导型一氧化氮合酶(iNOS)和环氧合酶-2(COX-2)的蛋白、mRNA表达,以及Toll样受体4(TLR4)mRNA表达。结果PGSF(10,1μmol·L^(-1))可明显降低LPS诱导的BV-2细胞培养基中TNF-α(P<0.01)和IL^(-1)β(P<0.05)浓度,同时下调细胞内iNOS蛋白(P<0.05)和mRNA(P<0.01,P<0.05)表达;PGSF(10μmol·L^(-1))可降低NO的浓度(P<0.05)、下调COX-2的蛋白和mRNA(P<0.05)表达,并逆转TLR4 mRNA表达的上调(P<0.05)。结论PGSF能够负调控LPS诱导的BV-2小胶质细胞炎性介质的释放和炎性蛋白酶的表达,抑制小胶质细胞激活,发挥对抗神经炎症的作用,此抗炎作用可能由TLR4受体介导。Objective To investigate the effect of polygalasaponin F（PGSF） on the activation of BV-2 microglial cells induced by lipopolysaccharide （LPS） and to explore its underlying mechanism. Methods Neuroinfiammatory model was established by exposure of BV-2 cells to LPS （0.1 μg·mL^-1） for 24 h. The concentrations of pro-inflammatory cytokines of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）, and nitric oxide （NO） content was determined by Griess method. The mRNA and protein levels of pro-inflammatory enzymes of inducible nitric oxide synthase （iNOS） and cyclooxygenase-2 （COX-2） were analyzed by reverse transcription-polymerase chain reaction （RT-PCR） and Western Blot respectively. RT-PCR was also used to detect the mRNA expression of inflammatory signal transducer of Toll-like receptor 4（TLR4）. Results The results indicated that PGSF （10, 1 μmol·L^-1） obviously inhibited the release of TNF-α （P 〈 0.01） and IL-1β（P 〈 0.05） in the BV-2 microglial ceils stimulated by LPS, and downregulated iNOS protein （P 〈 0.05） and mRNA （P 〈 0.01, P 〈 0.05） expression. PGSF （10μmol·L^-1） also reduced the generation of NO （P 〈 0.05 ） and downregulated COX-2 protein and mRNA （P 〈 0.05 ） expression in BV-2 cells stimulated by LPS. In addition, PGSF （10μmol·L^-1） reversed the increase of mRNA expression of TLR4（P 〈 0.05） in BV-2 cells stimulated by LPS. Conclusion PGSF can inhibit the activation of BV-2 microglial cells via downregulating the release of inflammatory mediators and expression of pro-inflammatory enzymes. TLR4 might be involved in the intracellular signal transduetion of anti-neuroinflammatory effect of PGSF.

关 键 词：瓜子金皂苷己 小胶质细胞 脂多糖 神经炎症 诱导型一氧化氮合酶 环氧合酶-2 TOLL样受体4 

分 类 号：R285.5[医药卫生—中药学]