Kalantar Kurosh, Gholijani Nasser, Mousaei Nashmin, Yazdani Malihe, Amirghofran Zahra
Department of Immunology, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.
Autoimmune Diseases Research Center and Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Antiinflamm Antiallergy Agents Med Chem. 2018;17(1):39-49. doi: 10.2174/1871523017666180608081656.
Dracocephalum kotschyi is traditionally used for its anti-inflammatory effects. We aimed to investigate the effects of ethyl acetate extract of D. kotschyi on the expression of key inflammatory mediators and main signaling molecules involved in the regulation of inflammation.
Lipopolysaccharide (LPS)-stimulated J774.1 mouse macrophages were cultured in the presence of the plant extract and examined by the real time-PCR for gene expressions of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Cytokine levels and phosphorylated forms of stressactivated protein kinases/c-Jun N-terminal kinase (SAPK/JNK), signal transducer and activator of transcription (STAT)-3, p38, IκB-α and nuclear factor (NF)-κB p65 were determined using ELISA.
The extract significantly reduced the expression of key mediators of inflammation. iNOS expression level decreased from 138±8.5 fold in LPS-only treated cells to 6.5±2.6 fold after treatment with 25 µg/ml of the extract (p<0.001). Similarly, COX-2 expression decreased from 632 ±98.8 fold in control to 124 ±24.6 fold (p<0.01). Treatment of cells with the extract significantly reduced IL-1β and TNF-α cytokines at both gene and protein expression levels. The extract at 25 µg/ml caused significant decreases in phospho- SAPK/JNK and phospho-STAT3 levels in macrophages (p<0.01). Proteins of phospho-p38, NFκB-p65 and phospho-NF-κB p65 had a reduced level in treated cells (p<0.05). No significant change in phospho-IκB level was observed.
These findings suggested that D. kotschyi with inhibition of NF-κB, SAPK/JNK, STAT-3 and p-38 might have reduced the expression levels of key inflammatory mediators and thus possibly have potential beneficial impact on inflammatory diseases.
传统上，科氏青兰因其抗炎作用而被使用。我们旨在研究科氏青兰乙酸乙酯提取物对关键炎症介质表达以及参与炎症调节的主要信号分子的影响。
在植物提取物存在的情况下培养脂多糖（LPS）刺激的J774.1小鼠巨噬细胞，并通过实时聚合酶链反应检测白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、诱导型一氧化氮合酶（iNOS）和环氧化酶（COX）-2的基因表达。使用酶联免疫吸附测定法测定细胞因子水平以及应激激活蛋白激酶/ c-Jun氨基末端激酶（SAPK/JNK）、信号转导和转录激活因子（STAT）-3、p38、IκB-α和核因子（NF）-κB p65的磷酸化形式。
提取物显著降低了关键炎症介质的表达。iNOS表达水平从仅用LPS处理的细胞中的13８±8.5倍降至用25μg/ml提取物处理后的6.5±2.6倍（p<0.001）。同样，COX-2表达从对照中的632±98.8倍降至124±24.6倍（p<0.01）。用提取物处理细胞在基因和蛋白质表达水平上均显著降低了IL-1β和TNF-α细胞因子。25μg/ml的提取物使巨噬细胞中的磷酸化SAPK/JNK和磷酸化STAT3水平显著降低（p<0.01）。处理细胞中磷酸化p38、NFκB-p65和磷酸化NF-κB p65的蛋白质水平降低（p<0.05）。未观察到磷酸化IκB水平有显著变化。
这些发现表明，抑制NF-κB、SAPK/JNK、STAT-3和p-38的科氏青兰可能降低了关键炎症介质的表达水平，因此可能对炎症性疾病具有潜在的有益影响。
