基于RNA-Seq数据分析葡萄糖诱导绵羊卵泡颗粒细胞衰老的差异表达基因

doi:10.16431/j.cnki.1671-7236.2022.02.025

Abstract

Abstract: 【Objective】 This study was to investigate the effect of glucose on the senescence and gene expression of sheep follicular granulosa cells.【Method】 Primary isolated sheep follicular granulosa cells were cultured in vitro with 17.5 (H group) and 2 mmol/L (L group) glucose.Each group was treated for 72 h, and cellular senescence was detected by β-galactoside staining.Transcriptome sequencing was performed on the preadipocytes by RNA-Seq technology, and the differentially expressed genes (DEGs) were analyzed by GO function enrichment analysis and KEGG signal pathway enrichment analysis.The expression levels of ankyrin repeat domain-containing protein 1(ANKRD1), interleukin-8(IL-8), oxytocin(OXT) and NADPH oxidase 4(NOX4) genes were detected by Real-time quantitative PCR.【Result】 The percentage of β-galactoside-positive cells in group H was extremely significantly higher than that in group L (P < 0.01).RNA-Seq results showed that there were 401 DEGs between the two groups, including 153 up-regulated genes and 248 down-regulated genes, including 9 genes related to cell abnormalities induced by high glucose and 6 genes related to cell cycle.The GO function enrichment analysis showed that the DEGs were involved in biological processes such as cellular process, biological regulation, metabolic process, multicellular organismal process and locomotion.The concentration of cellular component mainly included extracellular region, membrane, synapse, organelle and supramolecular complex.The enrichment of molecular function was mainly in catalytic activity, binding, transporter activity, molecular function regulator and structural molecule activity.KEGG signal pathway enrichment analysis showed that the DEGs were mainly concentrated in AGE-RAGE signaling pathway in diabetic complications, TNF signaling pathway, PPAR signaling pathway.The Real-time quantitative results showed that compared with L group, the expression level of IL-8 gene was extremely significantly up-regulated (P < 0.01), the expression level of ANKRD1 gene was significantly up-regulated (P < 0.05), and the expression level of OXT gene was extremely significantly down-regulated (P < 0.01), NOX4 gene expression showed an upward trend, but the difference was not significant (P > 0.05).The result of Real-time quantitative PCR was basically consistent with that of transcriptome sequencing.【Conclusion】 17.5 mmol/L glucose induced senescenc and genes expression changes in sheep follicular granulosa cells.The results provided a basis for the functional and molecular mechanism of glucose induced senescence of granulosa cells.

Key words: RNA-Seq; glucose; sheep; granulosa cells; senescence

CLC Number:

ZHANG Peiying, SONG Pengyan, ZHOU Ying, CHEN Xiaoyong, ZHOU Rongyan. Analysis of Differentially Expressed Genes in Glucose-induced Senescence of Sheep Follicular Granulosa Cells Based on RNA-Seq Data[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 631-640.

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Analysis of Differentially Expressed Genes in Glucose-induced Senescence of Sheep Follicular Granulosa Cells Based on RNA-Seq Data

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