Yang Yi, Guo Li, Wang Jin, Li WenYan, Zhou Xiaohong, Zhang Caiqun, Han Chenyang
Department of Pharmacy, The Second Affiliated Hospital of Jiaxing University, Hangzhou, China.
Department of Center Laboratory, The Second Affiliated Hospital of Jiaxing University, Hangzhou, China.
J Biochem Mol Toxicol. 2022 Jun;36(6):e23045. doi: 10.1002/jbt.23045. Epub 2022 Mar 15.
Arglabin (Arg) is a derivative of parthenolide. At present, there are few reports on the pharmacological effects and targets of Arg. In this study, we aimed to explore the relationship between Arg and NF-κB (P50) and the intervention effects of Arg on neuroinflammation. BV2 cells were cultured in vitro. LPS/IFN-γ was used to induce M1 polarization. After Arg intervention, the cytokine expression of M1 and M2 cell marker was detected, the expression of CD86 was detected by immunofluorescence (IF) staining, the levels of P50 and p-P50 were detected by Western blot and the expression of ROS was by DCFH-DA. AfterP50 knockout, we investigated the effect of P50 on the polarization of BV2 cells. Four-month-old APP/PS1 (AD) mice were treated with Arg by intragastric administration, followed by detection of the expression of CD86, CD206, and IBA-1 by IF staining, Finally, molecular-protein docking and Pull-down assays were used to validate the targeted binding relationship between P50 and Arg. Arg could inhibit the M1 polarization of BV2 cells, decrease the levels of TNF-α, IL-1β, IL-6, iNOS, and IL-12, and simultaneously inhibit the expression of P50 and p-P50. P50 knockout could inhibit the M1 polarization of BV2 cells, and P50 played an important role in the polarization of BV2 cells. Molecular docking and pull-down assays revealed that Arg and P50 had a targeted binding relationship. Animal experiments showed that Arg could regulate the polarization level of M1-M2 cells, increase the proportion of M2 cells, decrease the degree of nerve injury and suppress the expression of P50 and p-P50. In this study, we found that Arg could target P50 to regulate reprogramming of BV2 cells, inhibit M1 polarization, and increase the level of M2 cells, thereby exerting a neuroprotective effect.
阿格拉宾（Arg）是小白菊内酯的衍生物。目前，关于阿格拉宾的药理作用和靶点的报道较少。在本研究中，我们旨在探讨阿格拉宾与核因子κB（P50）之间的关系以及阿格拉宾对神经炎症的干预作用。体外培养BV2细胞。使用脂多糖/干扰素-γ诱导M1极化。阿格拉宾干预后，检测M1和M2细胞标志物的细胞因子表达，通过免疫荧光（IF）染色检测CD86的表达，通过蛋白质免疫印迹法检测P50和磷酸化P50（p-P50）的水平，通过2′,7′-二氯二氢荧光素二乙酸酯（DCFH-DA）检测活性氧（ROS）的表达。P50基因敲除后，我们研究了P50对BV2细胞极化的影响。对4月龄的淀粉样前体蛋白/早老素1（APP/PS1，阿尔茨海默病模型）小鼠进行阿格拉宾灌胃治疗，随后通过IF染色检测CD86、CD206和离子钙结合衔接分子1（IBA-1）的表达。最后，使用分子-蛋白质对接和下拉实验验证P50与阿格拉宾之间的靶向结合关系。阿格拉宾可抑制BV2细胞的M1极化，降低肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、诱导型一氧化氮合酶（iNOS）和白细胞介素-12的水平，同时抑制P50和p-P50的表达。P50基因敲除可抑制BV2细胞的M1极化，且P50在BV2细胞极化中起重要作用。分子对接和下拉实验表明阿格拉宾与P50存在靶向结合关系。动物实验表明，阿格拉宾可调节M1-M2细胞的极化水平，增加M2细胞比例，减轻神经损伤程度，并抑制P50和p-P50的表达。在本研究中，我们发现阿格拉宾可靶向P50来调节BV2细胞重编程，抑制M1极化，并提高M2细胞水平，从而发挥神经保护作用。
