白色紫锥菊不定根抑制LPS诱导的小鼠巨噬细胞促炎介质的产生

Adventitious roots of Echinacea pallida inhibit the production of pro-inflammatory mediators in LPS induced mouse macrophages

为探究白色紫锥菊不定根抗炎特性,采用MTT法测定白色紫锥菊不定根提取物的细胞毒性。小鼠腹腔中巨噬细胞用不定根提取物处理后,再用脂多糖(LPS)刺激,采用Griess试剂法测定一氧化氮(NO)含量,用ELISA试剂盒法测定前列腺素E2(PGE 2 )、白介素-6(IL-6)、白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平,用BCA试剂盒法测定一氧化氮合酶(iNOS)和环氧合酶-2(COX-2)蛋白含量。为研究丝裂原活化蛋白激酶(MAPKs)信号通路,用免疫印迹法测定MAPKs(p38、JKN和ERK)蛋白的磷酸化水平。结果表明:巨噬细胞中PGE 2 和NO的释放量在提取物浓度为50和100 μg/mL时显著降低,而IL-1β和TNF-α的释放量在25~100 μg/mL时有显著的抑制效果,但IL-6不受提取物的影响。且不定根提取物(25~100 μg/mL)显著抑制了p38、JKN和ERK蛋白的磷酸化。综上所述,白色紫锥菊不定根提取物具有显著的抗炎作用,并参与调控MAPKs信号通路。此研究结果为应用白色紫锥菊不定根生产抗炎相关产品提供参考依据。

To investigate the anti-inflammatory properties of adventitious roots of Echinacea pallida (Nutt.) Nutt., the cytotoxicity of the extract of E. pallida adventitious roots was determined by MTT assay. Macrophages in the peritoneal cavity of mice were treated with adventitious root extract and stimulated with lipopolysaccharide (LPS). After that, the content of nitric oxide (NO) was determined by Griess reagent method. The levels of prostaglandin E 2 (PGE 2), interleukin-6 (IL-6), interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were determined by ELISA kit. Meantime, the content of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein was determined by BCA kit . To study the mitogen-activated protein kinase (MAPKs) signaling pathway, phosphorylation levels of MAPKs (p38, JKN and ERK) proteins were determined by immunoblotting. The results showed that the release of PGE 2 and NO in macrophages was significantly inhibited at extract concentrations of 50 and 100 μg/mL. Also, the release of IL-1β and TNF-α was significantly inhibited at 25-100 μg/mL, but the release of IL-6 was not affected by the extract. In addition, the adventitious root extract (25-100 μg/mL) significantly inhibited the phosphorylation of mitogen-activated protein kinase (p38, JKN and ERK) proteins. In summary, the extract of E. pallida adventitious roots had an significant inhibitory effect on inflammation and participated in the regulation of MAPKs signaling pathway. The results in this study provided a reference for the application of E. pallida adventitious roots to produce anti-inflammatory related products.