Department of Cardiology, Affiliated Liyuan Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
School of Basic Medical Sciences, Hubei University of Chinese Medicine, Wuhan, China.
Immunopharmacol Immunotoxicol. 2020 Jun;42(3):286-293. doi: 10.1080/08923973.2020.1750628. Epub 2020 Apr 20.
Interleukin (IL)-33 has been attracting more and more attention as a new member of theIL-1 cytokine family in recent years. However, the underlying mechanisms referred to the regulation of endogenous IL-33 production are not fully illustrated. Paeoniflorin (PF) has been reported to possess multiple pharmacological activities, including anti-inflammation and anti-allergy. In this study, we aimed to investigate the effect of PF on IL-33 production by macrophages and explore the underlying mechanisms. , IL-33 production in mice after lipopolysaccharide (LPS) injection together with PF application was detected by enzyme-linked immunosorbent assay (ELISA). , MTT, Real-time PCR, ELISA, Calcium (Ca) imaging and Western blot were used to assess the cytotoxicity of PF, IL-33 expression at mRNA and protein levels, Ca influx, protein kinase C (PKC) activity, nuclear factor-kappa B (NF-κB), and mitogen-activated protein kinase (MAPK) activation in LPS-stimulated RAW264.7 macrophages with PF administration. Our results indicated that PF (5 and 25 mg/kg) significantly reduced the production of TNF-a, IL-1β, and IL-33 in the peritoneal exudate of LPS-treated mice. assay, upregulation of PF concentration (≥ 20 μM) showed an increased cytotoxicity in RAW264.7 cells during the 24-h cell culture. PF (10 μM) inhibited IL-33 production, Ca influx, PKC activity, NF-κB (p65) activation, and P38MAPK phosphorylation in LPS-treated macrophages. Notably, NF-κB inhibitor (BAY 11-7085), P38MAPK inhibitor (SB203580), and Ca blocker (NiCl) also curbed LPS-induced IL-33 production, respectively. PF suppresses IL-33 production by macrophages inhibiting NF-κB and P38MAPK activation associated with the regulation of Ca mobilization.
白细胞介素 (IL)-33 作为 IL-1 细胞因子家族的新成员，近年来受到越来越多的关注。然而，内源性 IL-33 产生的调节机制尚未完全阐明。丹皮酚 (PF) 具有多种药理活性，包括抗炎和抗过敏。在这项研究中，我们旨在研究 PF 对巨噬细胞中 IL-33 产生的影响，并探讨其潜在机制。通过酶联免疫吸附试验 (ELISA) 检测脂多糖 (LPS) 注射后 PF 处理对小鼠 IL-33 产生的影响。通过 MTT、实时 PCR、ELISA、钙 (Ca) 成像和 Western blot 评估 PF 对 RAW264.7 巨噬细胞的细胞毒性、IL-33 在 mRNA 和蛋白水平的表达、Ca 内流、蛋白激酶 C (PKC) 活性、核因子-κB (NF-κB) 和丝裂原活化蛋白激酶 (MAPK) 的激活作用。结果表明，PF（5 和 25mg/kg）显著降低 LPS 处理小鼠腹腔渗出液中 TNF-a、IL-1β 和 IL-33 的产生。在 24 小时细胞培养过程中，PF 浓度（≥20μM）上调显示 RAW264.7 细胞的细胞毒性增加。PF（10μM）抑制 LPS 处理的巨噬细胞中 IL-33 的产生、Ca 内流、PKC 活性、NF-κB（p65）激活和 P38MAPK 磷酸化。值得注意的是，NF-κB 抑制剂 (BAY 11-7085)、P38MAPK 抑制剂 (SB203580) 和 Ca 阻断剂 (NiCl) 也分别抑制 LPS 诱导的 IL-33 产生。PF 通过抑制 NF-κB 和 P38MAPK 激活以及调节 Ca 动员来抑制巨噬细胞中 IL-33 的产生。
