Department of Clinical Pharmacy, University of Medicine and Pharmacy, Vietnam National University, Hanoi, Vietnam.
College of Pharmacy, Gachon University, Incheon, Republic of Korea.
Pharm Biol. 2023 Dec;61(1):135-143. doi: 10.1080/13880209.2022.2159450.
Alkaloid-enriched extract of (Thunb.) Trevis (Lycopodiaceae) (HsAE) can potentially be used to manage neuronal disorders.
This study determines the anti-neuroinflammatory effects of HsAE on lipopolysaccharide (LPS)-stimulated BV-2 microglial cells and the underlying mechanisms.
BV-2 cells were pre- or post-treated with different concentrations of HsAE (25-150 µg/mL) for 30 min before or after LPS induction. Cell viability was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and no cytotoxicity was found. Nitric oxide (NO) concentration was determined using Griess reagent. The levels of prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were determined using enzyme-linked immunosorbent assay. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 and the phosphorylation of mitogen-activated protein kinase (MAPK) were analyzed using western blotting.
HsAE reduced LPS-induced NO production with half-maximal inhibitory concentration values of 99.79 and 92.40 µg/mL at pre- and post-treatment, respectively. Pre-treatment with HsAE at concentrations of 50, 100, and 150 µg/mL completely inhibited the secretion of PGE2, TNF-α, IL-6, and IL-1β compared to post-treatment with HsAE. This suggests that prophylactic treatment is better than post-inflammation treatment. HsAE decreased the expression levels of iNOS and COX-2 and attenuated the secretion of pro-inflammatory factors by downregulating the phosphorylation of p38 and extracellular signal-regulated protein kinase in the MAPK signaling pathway.
HsAE exerts anti-neuroinflammatory effects on LPS-stimulated BV-2 cells, suggesting that it may be a potential candidate for the treatment of neuroinflammation in neurodegenerative diseases.
（Thunb.）Trevis（石松科）的生物碱富集提取物（HsAE）可用于治疗神经紊乱。
本研究旨在确定 HsAE 对脂多糖（LPS）刺激的 BV-2 小胶质细胞的抗炎作用及其机制。
用不同浓度的 HsAE（25-150μg/mL）预处理或后处理 BV-2 细胞 30min，然后再用 LPS 诱导。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐法评估细胞活力，未发现细胞毒性。用 Griess 试剂测定一氧化氮（NO）浓度。酶联免疫吸附试验测定前列腺素 E2（PGE2）、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和 IL-6 的水平。用 Western blot 分析诱导型一氧化氮合酶（iNOS）和环氧化酶（COX）-2的水平以及丝裂原活化蛋白激酶（MAPK）的磷酸化。
HsAE 降低 LPS 诱导的 NO 生成，预治疗和后治疗的半最大抑制浓度值分别为 99.79 和 92.40μg/mL。与后治疗相比，浓度为 50、100 和 150μg/mL 的 HsAE 预处理完全抑制 PGE2、TNF-α、IL-6 和 IL-1β的分泌。这表明预防性治疗优于炎症后治疗。HsAE 通过下调 MAPK 信号通路中 p38 和细胞外信号调节蛋白激酶的磷酸化，降低 iNOS 和 COX-2 的表达水平，并减弱促炎因子的分泌。
HsAE 对 LPS 刺激的 BV-2 细胞具有抗炎作用，表明它可能是治疗神经退行性疾病中神经炎症的潜在候选药物。
