青蒿素调控NF-κB信号通路介导阿尔茨海默病炎症反应细胞模型的研究

Artemisinin Mediates Inflammatory Reaction in Alzheimer's Disease Cell Model by Regulating NF-κB Signaling Pathway

目的探讨青蒿素(ART)调控NF-κB通路介导阿尔茨海默病(AD)炎症反应细胞模型的作用及机制。方法建立AD炎症反应细胞模型。采用CCK8法检测ART对BV2小胶质细胞的作用浓度。采用Western blotting检测NF-κB信号通路蛋白(NF-κB P65,IKBα,p-NF-κB P65、p-IKBα)表达水平及炎症诱导酶环氧合酶2(COX-2)、诱导型一氧化氮合成酶(i NOS)的活化水平。实时PCR法检测促炎因子白细胞介素1β(IL-1β),肿瘤坏死因子α(TNF-α)及抑炎因子IL-10的m RNA表达水平。结果 Aβ25-35浓度为10μmol/L时,对BV2细胞生存率无显著影响。ART浓度在0.1~10μmol/L时细胞活力无显著变化,不具有细胞毒性。Western blotting结果显示,ART预处理组IKBα、NF-κB P65的蛋白表达水平升高,p-NF-k B P65和p-IKBα的蛋白表达水平显著降低,COX-2,i NOS的活化水平显著降低。实时PCR结果表明,ART预处理组促炎因子IL-1β、TNF-αm RNA的表达明显降低,而抑炎因子IL-10 m RNA的表达明显升高。结论 ART调控NF-κB信号通路,减少炎症诱导酶及调节炎性因子表达,发挥抗炎作用。

Objective To investigate the role and mechanism of artemisinin（ART） regulating NF-κB pathway in mediating inflammatory response in an Alzheimer＇s disease（AD） inflammatory cell model. Methods The AD inflammatory cell model was established. The CCK8 method was used to detect the concentration of ART on BV2 microglial cells. Western blotting was used to detect the expression levels of NF-κB signaling pathway proteins（NF-κB P65,IKBα,p-NF-κB P65,p-IKBα）,inflammation-inducing enzyme cyclooxygenase-2（COX-2）,and inducible ones. The level of nitric oxide synthase（iNOS） activation was determined. Real-time PCR was used to detect the expression levels of IL-1β,TNF-α,and IL-10. Results When Aβ25-35 concentration was 10 μmol/L,there was no significant effect on the survival rate of BV2 cells. When the concentration of ART was 0.1-10 μmol/L,there was no significant change in cell viability and no cytotoxicity. The results of Western blotting analysis showed that protein expression levels of IKBα and NF-κB P65 in the ART pretreatment group were increased,protein expression of p-NF-k B P65 and p-IKBα were significantly decreased,and activation of COX-2 and i NOS were significantly decreased. The results of real-time PCR showed that the expression of IL-1β and TNF-α m RNA was significantly decreased in the ART pretreatment group,whereas the expression of the IL-10 m RNA was significantly increased. Conclusion ART regulates the NF-κB signaling pathway,reduces inflammation-inducing enzymes,regulates the expression of inflammatory factors,and exerts anti-inflammatory effects.