Effects of EGCG on the expression of lncRNA in the longissimus dorsi muscle of Bama Xiang pig under heat stress
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Abstract
Objective This study aimed to investigate the effects of epigallocatechin-3-gallate (EGCG) on the expression of long non-coding RNA (lncRNA) in the longissimus dorsi muscle of Bama Xiang pigs under heat stress, with the goal of providing a theoretical basis for elucidating the molecular mechanism through which EGCG regulated intramuscular fat (IMF) deposition in Bama Xiang pigs under heat stress.Method Twenty male Bama Xiang pigs of 8-month-old were randomly assigned into four groups. The control group (TN) was reared at a normal temperature of 22 ℃, while the three high-temperature treatment groups (HS0, HS250, and HS500) were rear at 35 ℃. In the HS0, HS250, and HS500 groups, 0%, 0.025%, and 0.05% EGCG were added in their diets. At the end of the experiment, the Bama Xiang pigs were slaughtered and the longissimus dorsi muscle tissues were collected, from which the total RNA were extracted and their transcriptomes were sequenced. Differentially expressed lncRNAs were screened using DESeq to perform functional annotation analysis, KEGG signaling pathway enrichment analysis, and ceRNA interaction network construction. Five lncRNA target genes were randomly selected and analyzed using real-time fluorescence quantitative PCR to verify the reliability of the transcriptome sequencing results.Result A total of 263 differentially expressed lncRNAs were screened from the groups of TN vs HS0, TN vs HS500, and HS0 vs HS500, and 2819 target genes were enriched. Among them, the target genes PLPP1 of lncRNA MSTRG.11234, PPARα of ENSSSCT00000068789, and SCD5 of MSTRG.26033 were closely related to fat synthesis. KEGG signaling pathway enrichment analysis showed that, among the signaling pathways enriched by target genes of differentially expressed lncRNAs, peroxisome proliferator-activated receptors, adenosine monophosphate-activated protein kinase, and phosphatidylinositol 3-kinase-Akt pathways and so on were related to lipid metabolism. Results from the ceRNA interaction network revealed that some lncRNAs may regulate lipid metabolism and stress responses through ceRNA mechanisms. Five differentially expressed lncRNA target genes were randomly selected, and their expression trends were consistent with the transcriptome sequencing results, indicating that the transcriptome sequencing results were reliable.Conclusion 263 differentially expressed lncRNAs are screened, and their target genes were significantly enriched in lipid metabolism-related signaling pathways, including the peroxisome proliferator-activated receptor, adenosine monophosphate-activated protein kinase, and phosphatidylinositol 3-kinase-Akt pathways. PLPP1, PPARα, and SCD5 genes are closely associated with IMF deposition. Heat stress may mediate IMF deposition by regulating lncRNA expression and affect the transcriptional activity of downstream lipid metabolism-related genes.
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