Sarcodia suieae acetyl-xylogalactan regulate RAW 264.7 macrophage NF-kappa B activation and IL-1 beta cytokine production in macrophage polarization

猪肉丝虫乙酰木半乳聚糖调节 RAW 264.7 巨噬细胞 NF-κB 活化和巨噬细胞极化中 IL-1β 细胞因子的产生

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作者:Tsung-Meng Wu, Fan-Hua Nan, Kuan-Chu Chen, Yu-Sheng Wu

Abstract

In this study, the effects of acetyl-xylogalactan extracted from Sarcodia suieae on RAW 264.7 macrophage polarisation were evaluated. This extracted acetyl-xylogalactan had a monosaccharide composition of 91% galactose and 9% xylose, with polysaccharide and acetyl contents of 80.6% and 19.3%, respectively. MALDI-TOF mass spectrometry and NMR spectroscopy revealed the molecular weight of the acetyl-xylogalactan to be 88.5 kDa. After acetyl-xylogalactan treatment, RAW 264.7 macrophage polarisation was noted, along with enhanced phagocytic ability. Furthermore, the Cell Counting Kit-8 (CCK-8) assay was performed and the results demonstrated non-significant alteration in lactate dehydrogenase levels in the treated cells. Next, interleukin (IL) 1β, TNF, and Malt-1 expression in RAW 264.7 macrophages treated with the S. suieae acetyl-xylogalactan was investigated through real-time quantitative polymerase chain reaction, and the results demonstrated that S. suieae acetyl-xylogalactan induced IL-1β and Malt-1 expression. RNA sequencing analysis results indicated the S. suieae acetyl-xylogalactan positively regulated cytokine production and secretion, protein secretion, and response to IL-1 activation, based on the observed GO terms. The predicted target genes in the GO enrichment analysis were found to upregulate NF-κB signalling and M0 to M1 macrophage conversion through the observed cytokine production. Thus, acetyl-xylogalactan can positively regulate RAW 264.7 macrophage polarisation.

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