Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Sarcodia suieae Acetyl-xylogalactan Down-regulate RAW264.7 Macrophage Apoptosis Factors Ddit3 and Hyou1 Expression Under Heat Stress

Version 1 : Received: 23 July 2021 / Approved: 26 July 2021 / Online: 26 July 2021 (14:25:38 CEST)

How to cite: Wang, K.; Pan, P.; Wu, T.; Wu, Y. Sarcodia suieae Acetyl-xylogalactan Down-regulate RAW264.7 Macrophage Apoptosis Factors Ddit3 and Hyou1 Expression Under Heat Stress. Preprints 2021, 2021070581 (doi: 10.20944/preprints202107.0581.v1). Wang, K.; Pan, P.; Wu, T.; Wu, Y. Sarcodia suieae Acetyl-xylogalactan Down-regulate RAW264.7 Macrophage Apoptosis Factors Ddit3 and Hyou1 Expression Under Heat Stress. Preprints 2021, 2021070581 (doi: 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.

Keywords

Sarcodia suieae; Acetyl-xylogalactan; RAW264.7; Heat stress; Apoptosis

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