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目的研究海风藤提取物对不同聚集状态的β淀粉样蛋白1-42(Aβ1-42)诱导小胶质细胞中炎性因子白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)表达的影响。方法体外培养小鼠小胶质细胞株BV2,制备寡聚体和纤丝状Aβ1-42,应用四甲基偶氮唑蓝(MTT)比色法检测不同浓度的海风藤提取物对BV2细胞活力的影响;BV2细胞分为4个实验组,分别采用寡聚体Aβ1-42、寡聚体Aβ1-42+海风藤提取物、纤丝状Aβ1-42、纤丝状Aβ1-42+海风藤提取物干预48 h,并设空白对照组,应用实时荧光定量PCR测定各组IL-1β和TNF-α的表达水平。结果海风藤提取物在10～80 g/L范围内对细胞活性无影响;实时荧光定量结果显示,4个实验组的小胶质细胞表达IL-1β和TNF-α的水平均高于空白对照组。寡聚体Aβ1-42+海风藤提取物组IL-1β和TNF-α的表达水平低于寡聚体Aβ1-42组,纤丝状Aβ1-42+海风藤提取物组IL-1β和TNF-α的表达水平低于纤丝状Aβ1-42组。结论炎性指标IL-1β和TNF-α的表达增高证明寡聚体和纤丝状Aβ1-42均能激活小胶质细胞;IL-1β和TNF-α的表达降低,表明海风藤提取物可以抑制寡聚体和纤丝状Aβ1-42对小胶质细胞的激活。
Objective To investigate the effects of sea buckthorn extract on the expression of IL-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in microglial cells induced by β-amyloid 1-42 (Aβ 1-42) ) Expression. Methods Mouse microglia cell line BV2 was cultured in vitro and oligomers and fibrillar Aβ1-42 were prepared. MTT assay was used to detect the effects of different concentrations of sea buckthorn extract on BV2 cell viability BV2 cells were divided into four experimental groups, respectively, oligomeric Aβ1-42, oligomeric Aβ1-42 + sea buckthorn extractives, fibrillar Aβ1-42, filamentous Aβ1-42 + sea buckthorn extract Intervention 48 h, and a blank control group, the use of real-time fluorescent quantitative PCR determination of IL-1β and TNF-α expression levels. Results The extract of Piper kadsura had no effect on the cell viability in the range of 10 ~ 80 g / L. The real-time fluorescence quantitative results showed that the levels of IL-1β and TNF-α in the microglia of the four experimental groups were higher than that of the blank control group. The levels of IL-1β and TNF-α in oligomeric Aβ1-42 + Trifolium repens extract group were lower than those in oligomer Aβ1-42 group, fibrillar Aβ1-42 + Trifolium repens extract group, α was lower than fibrillar Aβ 1-42 group. Conclusion The increased expression of IL-1β and TNF-α, which are inflammatory markers, indicates that oligomers and fibrillar Aβ1-42 can activate microglial cells. The expression of IL-1β and TNF-α is decreased, Inhibition of microglia activation by oligomers and fibrillar Aβ1-42.