Miao Yu-Lu, He Pan, Zhang Wen-Xia, Zhang Wen-Zhi, Feng Min, Ni Yan
Shanxi Provincial Academy of Traditional Chinese Medicine Taiyuan 030012, China.
Shanxi University of Traditional Chinese Medicine Taiyuan 030619, China.
Zhongguo Zhong Yao Za Zhi. 2020 Feb;45(4):946-954. doi: 10.19540/j.cnki.cjcmm.20190619.502.
Based on 1H-NMR metabonomics technique and Western blot assay, the anti-inflammatory mechanism of Crepis crocea was discussed. In this study, male SD rats were treated with water extract(2.5 g·kg(-1)) and dexamethasone acetate(6.25×10(-4) g·kg(-1)) for one week, and the inflammation model was induced by lipopolysaccharide(LPS). Then the counts of inflammatory cells white blood ceel(WBC), eosinophil(EO), lymphocyte(LY), basophils(BA) and neutrophils(NE) in whole blood of rats were observed. The levels of serum inflammatory factors tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6 and the expression of nuclear factor-κB(NF-κB) signaling pathway p65 and p-IκBα proteins in lung tissues were detected, and the change rules of serum endogenous metabolites were analyzed by ~1H-NMR metabonomics technique. The levels of TNF-α, IL-1β, IL-6 and NF-κB signaling pathway p65 and p-IκBα proteins were combined with ~1H-NMR metabonomics to study the anti-inflammatory mechanism of C. crocea. The results showed that the water extract of C. crocea significantly decreased the number of WBC, NE, EO, increased the number of BA and LY, decreased the levels of TNF-α, IL-1β, IL-6 and the expression of p65 and p-IκBα protein in NF-κB signaling pathway, and effectively alleviated the inflammatory symptoms. In the correlation analysis of differential metabolites regulated of C. crocea, four significant metabolites were obtained, including glycine, creatine, methionine and succinic acid. The anti-inflammatory mechanism of C. crocea may be related to the decrease of TNF-α, IL-1β, IL-6 levels and the protein expression of NF-κB signaling pathway, as well as the regulation of glycine, creatine, methionine and succinic acid metabolism.
基于1H-NMR代谢组学技术和蛋白质免疫印迹法，探讨了黄鹤菜的抗炎机制。本研究中，雄性SD大鼠分别用水提取物（2.5 g·kg(-1)）和醋酸地塞米松（6.25×10(-4) g·kg(-1)）处理一周，并用脂多糖（LPS）诱导炎症模型。然后观察大鼠全血中炎症细胞白细胞（WBC）、嗜酸性粒细胞（EO）、淋巴细胞（LY）、嗜碱性粒细胞（BA）和中性粒细胞（NE）的计数。检测血清炎症因子肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、IL-6水平以及肺组织中核因子-κB（NF-κB）信号通路p65和p-IκBα蛋白的表达，并通过1H-NMR代谢组学技术分析血清内源性代谢物的变化规律。将TNF-α、IL-1β、IL-6水平以及NF-κB信号通路p65和p-IκBα蛋白与1H-NMR代谢组学相结合，研究黄鹤菜的抗炎机制。结果表明，黄鹤菜水提取物显著降低了WBC、NE、EO的数量，增加了BA和LY的数量，降低了TNF-α、IL-1β、IL-6水平以及NF-κB信号通路中p65和p-IκBα蛋白的表达，并有效缓解了炎症症状。在黄鹤菜调节的差异代谢物的相关性分析中，获得了4种显著的代谢物，包括甘氨酸、肌酸、蛋氨酸和琥珀酸。黄鹤菜的抗炎机制可能与TNF-α、IL-1β、IL-6水平的降低以及NF-κB信号通路的蛋白表达有关，还与甘氨酸、肌酸、蛋氨酸和琥珀酸代谢的调节有关。
