Wijatniko Bambang Dwi, Ishii Yoshiki, Hirayama Makoto, Suzuki Takuya
Graduate School of Integrated Sciences for Life, Hiroshima University, Higashihiroshima 739-8528, Japan.
Department of Food and Agricultural Product Technology, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia.
Foods. 2024 Oct 8;13(19):3198. doi: 10.3390/foods13193198.
The production of inflammatory cytokines such as tumor necrosis factor (TNF)-α by activated macrophage cells plays an important role in the development of intestinal inflammation. The present study investigated the anti-inflammatory effect of the protein hydrolysates prepared from the jack bean (JBPHs), (L.) DC, using the enzyme Alcalase, in a murine macrophage model, RAW 264.7 cells, which were stimulated by lipopolysaccharides. JBPHs reduced the TNF-α expression at the protein and mRNA levels through the downregulation of cellular signaling pathways involved in nuclear factor kappa B (NF-κB), extracellular signal-regulated kinase (ERK), and p38. A combination of mass spectrometry and in silico approaches identified 10 potential anti-inflammatory peptides in the JBPHs, including LFLLP and DFFL. Interestingly, while LFLLP targeted the NF-κB pathway, DFFL targeted p38 and ERK to suppress the TNF-α production in the RAW 264.7 cells. In addition, LFLLP and DFFL were localized in the cytosol of the cells. These results demonstrated that LFLLP and DFFL were incorporated by RAW 264.7 cells and, at least in part, contributed to the reduction in TNF-α by JBPHs. These peptides isolated from JBPHs may well be utilized as new alternatives to alleviate intestinal inflammation.
活化的巨噬细胞产生的炎性细胞因子，如肿瘤坏死因子（TNF）-α，在肠道炎症的发展中起重要作用。本研究在脂多糖刺激的小鼠巨噬细胞模型RAW 264.7细胞中，研究了用碱性蛋白酶从刀豆制备的蛋白水解物（JBPHs）的抗炎作用。JBPHs通过下调参与核因子κB（NF-κB）、细胞外信号调节激酶（ERK）和p38的细胞信号通路，在蛋白质和mRNA水平上降低了TNF-α的表达。质谱分析和计算机模拟方法相结合，在JBPHs中鉴定出10种潜在的抗炎肽，包括LFLLP和DFFL。有趣的是，LFLLP靶向NF-κB途径，而DFFL靶向p38和ERK以抑制RAW 264.7细胞中TNF-α的产生。此外，LFLLP和DFFL定位于细胞的细胞质中。这些结果表明，LFLLP和DFFL被RAW 264.7细胞摄取，并且至少部分地促成了JBPHs对TNF-α的降低作用。从JBPHs中分离出的这些肽很可能被用作减轻肠道炎症的新替代品。
