Department of Anesthesiology and Pain Medicine, Juntendo Tokyo Koto Geriatric Medical Center, 3-3-20 Shinsuna, Koto-ku, Tokyo 136-0075, Japan.
Department of Anesthesiology and Pain Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
Endocr Metab Immune Disord Drug Targets. 2020;20(7):1073-1080. doi: 10.2174/1871530320666200131103733.
Yokukansan is a traditional Japanese herbal medicine that has an antiallodynic effect in patients with chronic pain. However, the mechanisms by which yokukansan inhibits neuropathic pain are unclear.
This study aimed to investigate the molecular effects of yokukansan on neuroinflammation in U373 MG glioblastoma astrocytoma cells, which express a functional high-affinity neurokinin 1 receptor (substance P receptor), and produce interleukin (IL)-6 and IL-8 in response to stimulation by substance P (SP).
We assessed the effect of yokukansan on the expression of ERK1/2, P38 MAPK, nuclear factor (NF)-κB, and cyclooxygenase-2 (COX-2) in U373 cells by western blot assay. Levels of IL-6 and IL-8 in conditioned medium obtained after stimulation of cells with SP for 24 h were measured by enzyme-linked immunosorbent assay. All experiments were conducted in triplicate. Results were analyzed by one-way ANOVA, and significance was accepted at p < 0.05.
Yokukansan suppressed SP-induced production of IL-6 and IL-8 by U373 MG cells, and downregulated SP-induced COX-2 expression. Yokukansan also inhibited phosphorylation of ERK1/2 and p38 MAPK, as well as nuclear translocation of NF-κB, induced by SP stimulation of U373 MG cells.
Yokukansan exhibits anti-inflammatory activity by suppressing SP-induced production of IL-6 and IL-8 and downregulating COX-2 expression in U373 MG cells, possibly via inhibition of the activation of signaling molecules, such as ERK1/2, p38 MAPK, and NF-κB.
和汉三才散是一种传统的日本草药，对慢性疼痛患者具有抗痛觉过敏作用。然而，和汉三才散抑制神经病理性疼痛的机制尚不清楚。
本研究旨在探讨和汉三才散对表达功能性高亲和力神经激肽 1 受体（P 物质受体）的 U373 MG 神经胶质瘤星形胶质细胞中神经炎症的分子作用，该细胞在受到 P 物质（SP）刺激时会产生白细胞介素（IL）-6 和 IL-8。
我们通过 Western blot 检测法评估了和汉三才散对 U373 细胞中 ERK1/2、P38 MAPK、核因子（NF）-κB 和环氧化酶-2（COX-2）表达的影响。通过酶联免疫吸附试验测量细胞经 SP 刺激 24 小时后获得的条件培养基中 IL-6 和 IL-8 的水平。所有实验均重复进行 3 次。结果通过单因素方差分析进行分析，p<0.05 时认为差异有统计学意义。
和汉三才散抑制了 SP 诱导的 U373 MG 细胞产生 IL-6 和 IL-8，并下调了 SP 诱导的 COX-2 表达。和汉三才散还抑制了 SP 刺激 U373 MG 细胞后 ERK1/2 和 p38 MAPK 的磷酸化以及 NF-κB 的核转位。
和汉三才散通过抑制 SP 诱导的 U373 MG 细胞产生 IL-6 和 IL-8 以及下调 COX-2 表达来发挥抗炎作用，可能是通过抑制信号分子如 ERK1/2、p38 MAPK 和 NF-κB 的激活。
