樟芝多糖抵抗脂多糖诱导的神经细胞PC12炎症反应

Antrodia Camphorata Polysaccharide Inhibited the Inflammatory Response in PC12 Cell Induced by LPS

目的研究樟芝多糖(antrodia camphorata polysaccharide,APC)抵抗脂多糖(lipopolysaccharide,LPS)引起神经细胞PC12炎症反应的作用和机制。方法PC12细胞常规培养后,分为对照组、模型组、APC低剂量组、APC高剂量组,其中APC低剂量组采用10 mg·L^(-1) APC干预,APC高剂量组采用50 mg·L^(-1) APC干预。采用CCK-8法检测细胞存活率,光镜下观察细胞形态,激光共聚焦法分析观察Fluo3-AM染色后细胞内钙离子浓度,Western blot法检测TLR4、MD2以及下游MyD88的表达,ELISA法检测外分泌的TNF-α、IL-6和IL-1β的表达。RT-QPCR检测MD2、TLR4以及MyD88的mRNA表达。结果APC可以显著改善LPS诱导的神经炎症,APC组细胞存活率显著高于模型组,且高剂量组优于低剂量组。APC干预后,MD2、TLR4以及下游MyD88的表达显著下调,外分泌的TNF-α、IL-6和IL-1β表达亦下调,且APC高剂量组优于低剂量组。结论APC可以通过抑制TLR4-MD2及其下游因子表达抵抗LPS诱导的神经炎症,这为神经疾病的治疗提供了新的参考,为APC的开发应用提供借鉴。

OBJECTIVE To study the effect and mechanism of antrodia camphorata polysaccharide(APC)resistance to the inflammatory reaction of PC12cell which was induced by lipopolysaccharide(LPS).METHODS After routine culture,PC12 cell was divided into control group,model group,high and low dose of APC group.The low dose group was treated with 10mg-L-1APC.The high dose group was treated with 50mg-L-1APC.Cell viability was detected by CCK-8.Observated the cell morphology under light microscope,calcium intracellular ions detected by Fluo 3-AM fluorescence probe on Laser confocal microscope.The expression of TLR4,MD2 and downstream MyD88 were dected by Western blot.The expression of TNF-α, IL-6 and IL-1β dected by ELISA.The mRNA expression of MD2,TLR4 and MyD88 dected by RT-QPCR.RESULTS APC could significantly reduce the neuro inflammation induced by LPS.The cell survival rate in APC group was significantly higher than that in the model group,and the high dose group was better than the low dose group.After APC intervented,the expressions of MD2,TLR4 and downstream MyD88 were significantly down regulated,the expressions of TNF-α,IL-6and IL-1β were also down regulated,the high dose group was superior to the low dose group.CONCLUSION APC can resist the neuroinflammation induced by LPS.The mechanism is related to inhibiting the TLR4-MD2 and its downstream factors.It provides a new reference for the treatment of nerve diseases and application of APC.