基于RNA-Seq探讨玉竹多糖对D-半乳糖诱导衰老小鼠的保护作用

来源:用户上传      作者:何超平　彭莎　陈博威　李力松　廖端芳

　　 〔摘要〕 目的 探玉竹多糖对D-半乳糖诱导衰老小鼠认知功能保护作用的分子机制。方法 采用水提醇沉法提取玉竹总多糖组分并进行含量测定;将30只昆明小鼠随机分为正常组、模型组、玉竹多糖组（2 g/kg）。模型组及玉竹多糖组小鼠每日皮下注射D-半乳糖（150 mg/kg），正常组及模型组每日灌胃等体积生理盐水，同时玉竹多糖组小鼠灌胃等体积相应剂量的药物。给药8周后处死老鼠并收集海马组织，使用RNA测序（RNA-Seq）分析各组间差异表达的mRNAs，筛选玉竹多糖干预后表达逆转的mRNAs。通过基因本体分析及京都基因与基因组百科全书通路富集分析差异基因参与的主要生物学过程，实时荧光定量PCR（real-time quantitative PCR， RT-qPCR）验证测序结果。结果 玉竹多糖提取率为7.12%，含量为81.27%±0.02%。RNA-Seq分析显示玉竹多糖逆转了衰老小鼠海马组织19个mRNAs的表达，生物信息学分析显示这些基因可能通过各类代谢、血管内皮生长因子信号通路及过氧化物酶体增殖物激活受体信号通路发挥治疗作用，RT-qPCR验证了RNA-Seq结果中差异基因的准确性。结论 玉竹多糖可通过多条信号通路来保护D-半乳糖诱导的衰老小鼠的认知功能。
　　 〔关键词〕 玉竹多糖;衰老;RNA-Seq;认知障碍;D-半乳糖
　　 〔中图分类号〕R285.5 〔文献标志码〕A 〔文章编号〕doi：10.3969/j.issn.1674-070X.2022.03.004
　　〔Abstract〕 Objective To explore the molecular mechanism of Polygonatum odoratum polysaccharide on the cognitive function of D-galactose-induced aging mice. Methods The total polysaccharides of Polygonatum odoratum were extracted by water alcohol precipitation method and the content was determined. 30 KM mice were randomly divided into normal group， model group， and Polygonatum odoratum polysaccharide group. （2 g/kg）. Mice in the model group and Polygonatum odoratum polysaccharide group were injected subcutaneously with D-galactose （150 mg/kg） daily. At the same time， the normal group and model group were gavaged with equal volume of normal saline daily， and mice in the Polygonatum odoratum polysaccharide group were gavaged with equal volume of corresponding dose. After 8 weeks of administration， the mice were sacrificed and hippocampal tissues were collected. RNA sequencing （RNA-Seq） was used to analyze the differentially expressed mRNAs between the groups， and the mRNAs whose expression was reversed after the intervention of Polygonatum odoratum polysaccharide were screened. Through GO function and KEGG pathway enrichment analysis of the main biological processes involved in differential genes， the sequencing results real-time quantitative PCR （RT-qPCR）. Results The extraction rate of Polygonatum odoratum polysaccharide was 7.12%， and the content was 81.27%±0.02%， RNA-Seq analysis showed that Polygonatum odoratum polysaccharide reversed the expression of 19 mRNAs in the hippocampus of aging mice. Bioinformatics analysis showed that these genes may play a therapeutic role through various metabolism， vascular endothelial growth factor signaling pathway and PPAR signaling pathway. RT-qPCR verified the accuracy of RNA-Seq results. Conclusion Polygonatum odoratum polysaccharide may protect the cognitive function of D-galactose-induced aging mice through multiple signaling pathways.

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