Liu Qi, Gao Ming, Lyu Hong-Jun, Rao Zhi-Li, Zeng Nan
State Key Laboratory of Southwestern Chinese Medicine Resources Chengdu 611137, China College of Pharmacy, Chengdu University of Traditional Chinese Medicine Chengdu 611137, China.
Zhongguo Zhong Yao Za Zhi. 2021 Sep;46(18):4800-4807. doi: 10.19540/j.cnki.cjcmm.20210610.702.
This paper aimed to explore the anti-inflammatory effect of ethanol extract from Saposhnikoviae Radix in a lipopolysaccharide(LPS)-induced inflammation mouse model and its regulation of TLR4/NF-κB signaling pathway. The ethanol extract from Saposhnikoviae Radix was separated and purified on the macroporous adsorption resin and its main chemical components were identified by UPLC-QE/MS. The identification results showed that the top ten components of ethanol extract from Saposhnikoviae Radix were mainly chromones and coumarins. A mouse model of inflammation induced by intraperitoneal injection of LPS was used to investigate the anti-inflammatory effects of ethanol extract from Saposhnikoviae Radix after intragastric administration for seven successive days. Mice in all groups except for the control group were treated with intraperitoneal injection of LPS(0.015 g·kg~(-1)) one hour after the last administration, and twelve hours later, the blood was sampled and separated and the broncoalveolar lavage fluid(BALF) was collected. The levels of nitric oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β) in mouse serum and BALF were detected by ELISA. The harvested lung tissue was stained with hematoxylin-eosin(HE) for observing the pathological changes, followed by the detection of protein expression levels of related molecules in TLR4/NF-κB signaling pathway by Western blotting. The results showed that the ethanol extract from Saposhnikoviae Radix significantly ameliorated the pathological conditions in lung tissue of model mice, reversed the increase in NO, TNF-α, IL-6, and IL-1β levels of mouse serum and BALF, down-regulated the protein expression levels of Toll-like receptor 4(TLR4), myeloid differentiation factor(MyD88), and phosphorylated nuclear transcription factor κB-p65/nuclear transcription factor κB-p65(P-NF-κB p65/NF-κB p65), and up-regulated the NF-κB inhibitory protein α(IκBα). The ethanol extract from Saposhnikoviae Radix exhibited a good anti-inflammatory effect in the LPS-induced acute inflammation muse model, which might be related to the inhibition of the activation of TLR4/NF-κB inflammatory signaling pathway. Chromones and coumarins have been proved to be the active components for its anti-inflammatory effects.
本文旨在探讨防风乙醇提取物在脂多糖(LPS)诱导的炎症小鼠模型中的抗炎作用及其对TLR4/NF-κB信号通路的调控。采用大孔吸附树脂对防风乙醇提取物进行分离纯化,并用超高效液相色谱-四极杆飞行时间质谱(UPLC-QE/MS)鉴定其主要化学成分。鉴定结果表明,防风乙醇提取物的前十大成分主要为色酮类和香豆素类。采用腹腔注射LPS诱导的小鼠炎症模型,连续7天灌胃给予防风乙醇提取物,观察其抗炎作用。除对照组外,其余各组小鼠在末次给药1小时后腹腔注射LPS(0.015 g·kg⁻¹),12小时后采集血液并分离血清,收集支气管肺泡灌洗液(BALF)。采用酶联免疫吸附测定(ELISA)法检测小鼠血清和BALF中一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的水平。取收获的肺组织进行苏木精-伊红(HE)染色,观察病理变化,然后采用蛋白质印迹法检测TLR4/NF-κB信号通路相关分子的蛋白表达水平。结果表明,防风乙醇提取物能显著改善模型小鼠肺组织的病理状况,降低小鼠血清和BALF中NO、TNF-α、IL-6和IL-1β的水平,下调Toll样受体4(TLR4)、髓样分化因子(MyD88)和磷酸化核转录因子κB-p65/核转录因子κB-p65(P-NF-κB p65/NF-κB p65)的蛋白表达水平,上调NF-κB抑制蛋白α(IκBα)。防风乙醇提取物在LPS诱导的急性炎症小鼠模型中具有良好的抗炎作用,其机制可能与抑制TLR4/NF-κB炎症信号通路的激活有关。色酮类和香豆素类已被证明是其抗炎作用的活性成分。