基于RNA-Seq探討玉竹多糖對D-半乳糖誘導衰老小鼠的保護作用

何超平　彭莎　陳博威　李力松　廖端芳

〔摘要〕 目的 探討玉竹多糖對D-半乳糖誘導衰老小鼠認知功能保護作用的分子機制。方法 采用水提醇沉法提取玉竹總多糖組分并進行含量測定;將30只昆明小鼠隨機分為正常組、模型組、玉竹多糖組（2 g/kg）。模型組及玉竹多糖組小鼠每日皮下注射D-半乳糖（150 mg/kg），正常組及模型組每日灌胃等體積生理鹽水，同時玉竹多糖組小鼠灌胃等體積相應劑量的藥物。給藥8周后處死老鼠并收集海馬組織，使用RNA測序（RNA-Seq）分析各組間差異表達的mRNAs，篩選玉竹多糖干預后表達逆轉的mRNAs。通過基因本體分析及京都基因與基因組百科全書通路富集分析差異基因參與的主要生物學過程，實時熒光定量PCR（real-time quantitative PCR， RT-qPCR）驗證測序結果。結果 玉竹多糖提取率為7.12%，含量為81.27%±0.02%。RNA-Seq分析顯示玉竹多糖逆轉了衰老小鼠海馬組織19個mRNAs的表達，生物信息學分析顯示這些基因可能通過各類代謝、血管內皮生長因子信號通路及過氧化物酶體增殖物激活受體信號通路發揮治療作用，RT-qPCR驗證了RNA-Seq結果中差異基因的準確性。結論 玉竹多糖可通過多條信號通路來保護D-半乳糖誘導的衰老小鼠的認知功能。

〔關鍵詞〕 玉竹多糖;衰老;RNA-Seq;認知障礙;D-半乳糖

〔中圖分類號〕R285.5? ? ? ?〔文獻標志碼〕A? ? ? ? 〔文章編號〕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.

〔Keywords〕 Polygonatum odoratum polysaccharid; aging; RNA-Seq; cognitive impairment; D-galactose

隨著人口老齡化的不斷加劇，衰老相關疾病發病率也逐漸增高[1]。衰老是機體各組織、器官功能隨年齡增長而發生的退行性變化，其中神經退行性病變是指由于神經元進行性變性死亡導致的以大腦為主的中樞神經系統損害的神經系統疾病[2]。……