Wang, K. T., Pan, P. K., Wu, T. M., & Wu, Y. S. (2021). Sarcodia suieae Acetyl-xylogalactan Down-regulate RAW264.7 Macrophage Apoptosis Factors Ddit3 and Hyou1 Expression Under Heat Stress. Preprints. https://doi.org/10.20944/preprints202107.0581.v1
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Wang, K., Tsung-Meng Wu and Yu-Sheng Wu. 2021 "Sarcodia suieae Acetyl-xylogalactan Down-regulate RAW264.7 Macrophage Apoptosis Factors Ddit3 and Hyou1 Expression Under Heat Stress" Preprints. https://doi.org/10.20944/preprints202107.0581.v1
Abstract
We aimed to evaluate the protective effects of acetyl-xylogalactan on the activity of RAW264.7 macrophages against heat stress. To this end, we assessed cell survival, phagocytic activity, in-tracellular Ca2+ level, mitochondria potential exchange, apoptotic assay findings, galactosidase activity and the RNA-seq by NGS and real-time polymerase chain reaction (PCR) expression. In our evaluation of macrophage morphology at 37°C and 41°C, the macrophages showed an oval shape at 41°C , unlike the spindle shape at 37°C. Therefore, 41°C was chosen as the heat stress condition. Subsequently, we designed an experiment to evaluate changes in the RAW264.7 macrophages after acetyl-xylogalactan treatment under heat stress. The survival of RAW264.7 macrophages treated with acetyl-xylogalactan was higher than that of controls that were not treated with acetyl-xylogalactan. Moreover, on the basis of the results of the annexin-V detection assay, the apoptotic activity of macrophages appeared to have reduced after treatment with ac-etyl-xylogalactan. Moreover, treatment with acetyl-xylogalactan resulted in a stronger recovery trend in the intracellular Ca2+ and mitochondrial membrane potential after heat stress. RNA sequencing and real-time polymerase chain reaction (PCR) illustrated that Sarcodia suieae acetyl-xylogalactan could upregulate the expression of the anti-apoptosis Cflar gene and down-regulate the expression of the apoptosis factors Ddit3, and Hyou1 to protect macrophages under heat stress.
Biology and Life Sciences, Biochemistry and Molecular Biology
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