Li Li, Liu Ru, He Jing, Li Jing, Guo Juan, Chen Yun, Ji Ke
Department of Psychiatry, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China.
Department of Psychiatry, Hubei Institute of Traditional Chinese Medicine, Wuhan 430074, China.
Evid Based Complement Alternat Med. 2022 May 19;2022:3492058. doi: 10.1155/2022/3492058. eCollection 2022.
Microglial BV-2 cells are activated in the brain following insomnia. Naringin (NAR) is a polymethoxylated flavonoid that is also commonly found in citrus fruits and is known for its antioxidant potential. However, the effect of NAR on microglial cells has rarely been studied in the brain of an organism after insomnia. This study aimed to investigate the effects and potential mechanisms of action of NAR on microglial cell activation and inflammation.
BV-2 cells were obtained from the China Center for Type Culture Collection and randomly divided into five treatment groups: control, model, NAR (10 M), WP1066 (5 M), and NAR + WP1066. With the exception of the control group, all groups were stimulated with LPS (1 g/mL) for 6 h. CCK8 was used to quantify cell viability and a scratch test was performed to detect cell migration. The expression levels of interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-), interleukin 1 beta (IL-1), nterleukin 10 (IL-10), and insulin like growth factor (1IGF-1) were measured by ELISA. Western blotting was performed to determine the levels of p-STAT3 and p-JAK. The Focalcheck™ Thin-Ring Fluorescent Microspheres kit was used to detect cell phagocytosis. Immunofluorescence was used to observe the expression of iNOS and arginase1 in BV-2 cells.
Compared with the control group, cell migration, cell viability, and the expression of IL-1, IL-6, TNF-, and iNOS were significantly increased in the model group, whereas the expression levels of IL-10, IGF-1, and arginase 1, as well as cell phagocytosis were reduced. With the increase in NAR concentration, cell migration, cell viability, the expression levels of IL-1, IL-6, TNF-, and iNOS decreased, while the expression of IL-10, IGF-1, and arginase 1 increased. Compared with the control group, p-STAT3, and p-JAK expression in the model group were significantly increased (<0.05). Compared with the model group, the expression of p-STAT3 and p-JAK in the NAR, NAR + WP1066, and WP1066 groups was significantly decreased ( < 0.05).
NAR treatment inhibited the proliferation, migration, and inflammation of BV-2 cells as well as the activation of microglia to the M1 phenotype. Conversely, NAR treatment promoted the activation of microglia to the M2 phenotype and enhanced the phagocytic function of BV-2 cells by regulating the activity of the JAK/STAT3 pathway.
失眠后大脑中的小胶质细胞BV-2会被激活。柚皮苷（NAR）是一种多甲氧基黄酮类化合物，常见于柑橘类水果中，以其抗氧化潜力而闻名。然而，在失眠后的生物体大脑中，NAR对小胶质细胞的影响鲜有研究。本研究旨在探讨NAR对小胶质细胞激活和炎症的影响及潜在作用机制。
从中国典型培养物保藏中心获得BV-2细胞，并随机分为五个处理组：对照组、模型组、NAR（10μM）组、WP1066（5μM）组和NAR + WP1066组。除对照组外，所有组均用脂多糖（1μg/mL）刺激6小时。使用CCK8定量细胞活力，并进行划痕试验检测细胞迁移。通过酶联免疫吸附测定法（ELISA）测量白细胞介素6（IL-6）、肿瘤坏死因子-α（TNF-α）、白细胞介素1β（IL-1）、白细胞介素10（IL-10）和胰岛素样生长因子（IGF-1）的表达水平。进行蛋白质免疫印迹法以确定p-STAT3和p-JAK的水平。使用Focalcheck™ 薄环荧光微球试剂盒检测细胞吞噬作用。采用免疫荧光法观察BV-2细胞中诱导型一氧化氮合酶（iNOS）和精氨酸酶1的表达。
与对照组相比，模型组细胞迁移、细胞活力以及IL-1、IL-6、TNF-α和iNOS的表达显著增加，而IL-10、IGF-1和精氨酸酶1的表达水平以及细胞吞噬作用降低。随着NAR浓度的增加，细胞迁移、细胞活力、IL-1、IL-6、TNF-α和iNOS的表达水平降低，而IL-10、IGF-1和精氨酸酶1的表达增加。与对照组相比，模型组中p-STAT3和p-JAK表达显著增加（P<0.05）。与模型组相比，NAR组、NAR + WP1066组和WP1066组中p-STAT3和p-JAK的表达显著降低（P<0.05）。
NAR处理可抑制BV-2细胞的增殖、迁移和炎症以及小胶质细胞向M1表型的激活。相反，NAR处理通过调节JAK/STAT3途径的活性促进小胶质细胞向M2表型的激活，并增强BV-2细胞的吞噬功能。
