| Acrolein is a highly reactive α-β unsaturated aldehyde,which is a colorless,flammable,volatile,pure liquid known for its pungent odor and strong irritation to mucous membranes,especially the eyes and upper respiratory tract.Thermal processing of foods is an important source of acrolein in the atmosphere.Acrolein is produced by various pathways during thermal processing of foods and is widely distributed in fried baked goods,overheated vegetable oils,alcoholic beverages,and foods rich in lipids and carbohydrates.Acrolein is neurotoxic,genotoxic and potentially carcinogenic;therefore,the search for active substances that can reduce or antagonize the toxic effects of acrolein in foods is of great importance to protect humans from its hazards.Ganoderma atrum(G.atrum)has been touted for thousands of years as a medicine or diet with high nutritional value.Ganoderma atrum polysaccharides(PSG-1)exhibits a wide range of beneficial,health effects,including immunomodulatory,chemo-protective,antioxidant and hypoglycemic activities.In particular,recent studies have also demonstrated that PSG-1 can modulate mucosal immunity through epithelial cells.However,little information is available on acrolein-induced effects on the intestine.Therefore,in this study,we established a model of acrolein-induced IEC-6 cell injury,and then comprehensively screened PSG-1 and acrolein-treated or untreated IEC-6 transcripts by whole-transcriptome sequencing.The main findings are summarized as follows:1.Acrolein applied to IEC-6 cells could reduce cell viability,disrupt intracellular oxidative homeostasis,decrease SOD activity and GPx activity,increase MDA content,and decrease ocdcludin,claudin and ZO-1 protein levels;could increase autophagy key protein LC3 and Beclin 1 protein levels,decrease p-m TOR and p-akt expression levels,thus activating the downstream pathway of autophagy;could reduce MMP,decrease Bcl-2 protein expression and increase the expression levels of apoptosis key proteins Caspase-3 and Caspase-9,thus inducing apoptosis.2.PSG-1 restored acrolein-induced oxidative damage,increased the antioxidant capacity of IEC-6 cells,enhanced acrolein-reduced expression of ocludin,claudin and ZO-1,and maintained the integrity of intestinal mucosa.PSG-1 treatment reduced autophagy key protein overexpression and slowed down acrolein-induced pleiotropic autophagy.PSG-1 treatment of cells increased MMP,increased Bcl-2 protein expression,decrease apoptosis key protein Caspase-3 and Caspase-9 expression levels,and prevent endogenous pathway of apoptosis thereby protecting cells.3.The expression of tight junction proteins increased after the action of autophagy inhibitor and apoptosis inhibitor on cells IEC-6;the expression of autophagy protein LC3 was reduced in IEC-6 cells treated with autophagy inhibitor,and the expression of Bcl-2,Caspase-3 and Caspase-9 was inhibited by autophagy inhibitor;the expression of Caspase-3 protein was reduced in IEC-6 cells treated with apoptosis inhibitor,and the expression of key autophagy protein Beclin 1 was inhibited by autophagy inhibitor.4.By whole transcriptome sequencing,the obtained m RNA data showed that compared with the control group,238 differentially expressed genes were screened in the acrolein group,including 142 up-regulated genes and 96 down-regulated genes;512 differentially expressed genes were screened in the PSG-1 group,including 262up-regulated genes and 250 down-regulated genes;compared with the acrolein group,a total of 327 differentially expressed.In comparison with the acrolein group,327 differentially expressed genes were screened in the PSG-1 group,including 195up-regulated genes and 132 down-regulated genes.For the resulting DEGs,GO and KEGG enrichment analyses were performed,and the first three enriched most significant functional sets were oxidoreductase activity,regulation of cytokine production,and response to extracellular stimuli.KEGG enrichment analysis identified cytokine-cytokine receptor interactions,glutathione metabolism and other pathways. |
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