Department of Pathology, College of Korean Medicine, Gachon University, Seongnam 13120, Republic of Korea.
Department of Medical Classics and History, College of Korean Medicine, Gachon University, Seongnam 13120, Republic of Korea.
Int J Mol Sci. 2023 Jan 20;24(3):2062. doi: 10.3390/ijms24032062.
Moutan Cortex, root, has long been used as a medicine for the treatment of inflammatory diseases. The aim of this study was to evaluate the modulative properties of Moutan Cortex water extract (CP) on endoplasmic reticulum (ER) stress-related macrophage activation via the calcium-CHOP pathway. RAW 264.7 mouse macrophages were activated by lipopolysaccharide (LPS), and the levels of various inflammatory mediators from RAW 264.7 were evaluated. The multiplex cytokine assay was used to investigate both cytokines and growth factors, and RT-PCR was used to investigate the expressions of inflammation-related genes, such as CHOP. Data represent the levels of NO and cytosolic calcium in LPS-stimulated RAW 264.7 were significantly inhibited by CP as well as hydrogen peroxide ( < 0.05). Minutely, NO production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 97.32 ± 1.55%, 95.86 ± 2.26%, 94.64 ± 1.83%, and 92.69 ± 2.31% of the control value (LPS only), respectively ( < 0.05). Calcium release in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 18 h was 95.78 ± 1.64%, 95.41 ± 1.14%, 94.54 ± 2.76%, and 90.89 ± 3.34% of the control value, respectively ( < 0.05). Hydrogen peroxide production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 79.15 ± 7.16%, 63.83 ± 4.03%, 46.27 ± 4.38%, and 40.66 ± 4.03% of the control value, respectively ( < 0.05). It is interesting that the production of IL-6, TNF-α, G-CSF, MIP-1α, MIP-2, and M-CSF in LPS-stimulated RAW 264.7 were significantly inhibited by CP ( < 0.05), while the production of LIX, LIF, RANTES, and MIP-1β showed a meaningful decrease. CP at concentrations of 25, 50, 100, and 200 µg/mL significantly reduced the transcription of , α, , , , , , , , , , and in LPS-stimulated RAW 264.7 ( < 0.05). CP at concentrations of 25, 50, and 100 µg/mL significantly reduced the phosphorylation of STAT3, p38 MAPK, and IκB-α in LPS-stimulated RAW 264.7 ( < 0.05). These results suggest that CP might modulate macrophage activation via LPS-induced calcium signaling and the ER stress-CHOP pathway.
丹皮，根，长期以来一直被用作治疗炎症性疾病的药物。本研究旨在通过钙-CHOP 途径评估丹皮水提取物 (CP) 对内质网 (ER) 应激相关巨噬细胞激活的调节作用。用脂多糖 (LPS) 激活 RAW 264.7 小鼠巨噬细胞，评估来自 RAW 264.7 的各种炎症介质的水平。使用多重细胞因子分析来研究细胞因子和生长因子，并用 RT-PCR 来研究炎症相关基因，如 CHOP 的表达。数据表示 LPS 刺激的 RAW 264.7 中的 NO 和胞质钙水平显著被 CP 和过氧化氢 ( < 0.05) 抑制。更具体地说，用浓度为 25、50、100 和 200 µg/mL 的 CP 孵育 24 小时的 LPS 刺激的 RAW 264.7 中 NO 的产生分别为对照值 (仅 LPS) 的 97.32 ± 1.55%、95.86 ± 2.26%、94.64 ± 1.83%和 92.69 ± 2.31% ( < 0.05)。用浓度为 25、50、100 和 200 µg/mL 的 CP 孵育 18 小时的 LPS 刺激的 RAW 264.7 中的钙释放分别为对照值的 95.78 ± 1.64%、95.41 ± 1.14%、94.54 ± 2.76%和 90.89 ± 3.34% ( < 0.05)。用浓度为 25、50、100 和 200 µg/mL 的 CP 孵育 24 小时的 LPS 刺激的 RAW 264.7 中的过氧化氢产生分别为对照值的 79.15 ± 7.16%、63.83 ± 4.03%、46.27 ± 4.38%和 40.66 ± 4.03% ( < 0.05)。有趣的是，CP 显著抑制了 LPS 刺激的 RAW 264.7 中 IL-6、TNF-α、G-CSF、MIP-1α、MIP-2 和 M-CSF 的产生 ( < 0.05)，而 LIX、LIF、RANTES 和 MIP-1β 的产生则呈显著下降趋势。浓度为 25、50、100 和 200 µg/mL 的 CP 显著降低了 LPS 刺激的 RAW 264.7 中转录因子 的转录 ( < 0.05)。浓度为 25、50 和 100 µg/mL 的 CP 显著降低了 LPS 刺激的 RAW 264.7 中 STAT3、p38 MAPK 和 IκB-α 的磷酸化 ( < 0.05)。这些结果表明 CP 可能通过 LPS 诱导的钙信号和内质网应激-CHOP 途径来调节巨噬细胞的激活。
