Wang Yuyin, Wei Wenyue, Guo Minfang, Li Suyao, Chai Zhi, Ma Cungen, Jiang Yuqiang, Song Lijuan, Yu Jiezhong
Key Laboratory of State Administration of Traditional Chinese Medicine for Multiple Sclerosis Treatment by Enriching Qi and Promoting Blood Circulation, Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619; Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases, Institute of Brain Science, Shanxi Datong University, Datong 037009, China.
Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases, Institute of Brain Science, Shanxi Datong University, Datong 037009; Dept. of Neurology, First Affiliated Hospital, Shanxi Medical University, Taiyuan 030001, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Dec;37(12):1066-1072.
Objective To investigate the effect of Lycium barbarum polysaccharide (LBP) on the polarization of BV2 microglia from M1 to M2 induced by lipopolysaccharide (LPS) and its mechanism. Methods The BV2 microglia were divided into control group, LPS group, and LBP treatment group (0.6, 0.9, 1.2) g/L. MTT assay was used to observe the cell viability of BV2 cells, and Griess assay was used to detect the release of NO. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISA. The expressions of Toll-like receptor 4 (TLR4), nuclear factor kappa B (NF-κB), inducible nitric oxide synthase (iNOS), and arginase-1 (Arg1) were detected by immunofluorescence cytochemistry. Western blot was used to evaluate the protein levels of ionized calcium-binding adaptor molecule-1 (Iba-1), TLR4, NF-κB, iNOS, and Arg1. Results There was no significant difference of the cell survival rate after treatment with different doses of LBP. Compared to those in the control group, in LPS group the BV2 microglia were activated with amoeba-like shape and increased release of NO, the expressions of Iba-1, TLR4, NF-κB, iNOS, TNF-α, IL-1β, and IL-6 were significantly increased, while the expressions of Arg1 and IL-10 was significantly decreased. In LBP group, Iba-1, TLR4, NF-κB, iNOS, TNF-α, IL-1β, and IL-6 were dramatically decreased and negatively correlated with the dose, while Arg1 and IL-10 were increased and positively correlated with the dose. Conclusion LBP inhibits activation of BV2 microglia induced by LPS and promots the M2 polarization, which may be realized through inhibiting TLR4/NF-κB signaling pathway.
目的 探讨枸杞多糖（LBP）对脂多糖（LPS）诱导的BV2小胶质细胞从M1型向M2型极化的影响及其机制。方法 将BV2小胶质细胞分为对照组、LPS组和LBP处理组（0.6、0.9、1.2）g/L。采用MTT法观察BV2细胞的活力，采用Griess法检测NO的释放。通过ELISA检测肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的水平。采用免疫荧光细胞化学法检测Toll样受体4（TLR4）、核因子κB（NF-κB）、诱导型一氧化氮合酶（iNOS）和精氨酸酶-1（Arg1）的表达。采用蛋白质印迹法评估离子钙结合衔接分子-1（Iba-1）、TLR4、NF-κB、iNOS和Arg1的蛋白水平。结果 不同剂量LBP处理后细胞存活率无显著差异。与对照组相比，LPS组BV2小胶质细胞呈阿米巴样活化，NO释放增加，Iba-1、TLR4、NF-κB、iNOS、TNF-α、IL-1β和IL-6的表达显著增加，而Arg1和IL-10的表达显著降低。在LBP组中，Iba-1、TLR4、NF-κB、iNOS、TNF-α、IL-1β和IL-6显著降低且与剂量呈负相关，而Arg1和IL-10增加且与剂量呈正相关。结论 LBP抑制LPS诱导的BV2小胶质细胞活化并促进M2极化，这可能是通过抑制TLR4/NF-κB信号通路实现的。
