Kim Ji Won, Kwon Ki Woong, Kim Mi-Yeon, Cho Jae Youl
Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Korea.
School of Systems Biomedical Science, Soongsil University, Seoul 06978, Korea.
Plants (Basel). 2022 Jun 30;11(13):1750. doi: 10.3390/plants11131750.
Inflammation is an immune response that protects against harmful stimuli. However, severe inflammation can cause many diseases, such as diabetes, cancer, and arthritis. In this study, we examined the anti-inflammatory efficacy and mechanism of Nutt. ethanol extract (Pp-EE) as a new strategy for controlling the inflammatory response. Cellular activities and the molecular target of Pp-EE were identified in RAW264.7 cells and HEK293T cells. The effect of Pp-EE was analyzed using the Griess assay, the luciferase assay, reverse transcription-polymerase chain reaction, and Western blotting. To evaluate the in vivo effects, an HCl/EtOH-induced gastritis mouse model was used. NO production and pro-inflammatory gene (, , and ) mRNA levels were decreased by Pp-EE in a concentration-dependent manner without showing cytotoxicity. The activation of the transcription factor, particularly NF-κB, was effectively suppressed by Pp-EE. It was also found that Pp-EE directly inhibits the activation of Src in lipopolysaccharide (LPS)-treated RAW264.7 cells and in Src-overexpressed HEK293 cells by Western blotting analysis and cellular thermal shift assay. Experiments in the gastritis mouse model indicated that Pp-EE suppresses HCl/EtOH-induced gastric lesions, the expression levels of , , and , and the phosphorylation of p65, p50, and Src. Taken together, these results suggest that Pp-EE can be applied as an anti-inflammatory remedy with a Src/NF-κB inhibitory property.
炎症是一种针对有害刺激的免疫反应。然而,严重的炎症会引发许多疾病,如糖尿病、癌症和关节炎。在本研究中,我们研究了坚果乙醇提取物(Pp-EE)作为控制炎症反应的新策略的抗炎功效及机制。在RAW264.7细胞和HEK293T细胞中确定了Pp-EE的细胞活性和分子靶点。使用格里斯试剂法、荧光素酶测定法、逆转录-聚合酶链反应和蛋白质印迹法分析了Pp-EE的作用。为了评估体内效果,使用了盐酸/乙醇诱导的胃炎小鼠模型。Pp-EE以浓度依赖性方式降低了一氧化氮的产生和促炎基因( 、 和 )的mRNA水平,且未显示出细胞毒性。Pp-EE有效抑制了转录因子尤其是核因子κB的激活。通过蛋白质印迹分析和细胞热迁移分析还发现,Pp-EE直接抑制脂多糖(LPS)处理的RAW264.7细胞和Src过表达的HEK293细胞中Src的激活。胃炎小鼠模型实验表明,Pp-EE可抑制盐酸/乙醇诱导的胃部损伤、 、 和 的表达水平以及p65、p50和Src的磷酸化。综上所述,这些结果表明Pp-EE可作为一种具有Src/核因子κB抑制特性的抗炎药物应用。