基于转录组学探索蒸制黄精多糖防治血虚模型小鼠的作用机制

Explore the Mechanism of Steam-processed Polygonatum Sibiricum Polysaccharides in Prophylaxis and Treatment of Blood Deficiency Mice Model Based on Transcriptomics

目的采用转录组测序技术(RNA sequencing,RNA-seq)探索蒸制黄精多糖防治小鼠血虚证(blood deficiency syndrome,BDS)的作用机制。方法将小鼠随机分为5组(每组10只),即正常组、模型组、黄精多糖组(0.1,0.4 g·kg^(-1))、当归补血口服液阳性药组。采用乙酰苯肼和环磷酰胺诱导建立BDS小鼠模型。连续给药14 d后,测定小鼠血常规、体质量和体温;采用RNA-seq技术对BDS小鼠肝组织进行转录组测序分析,筛选出黄精多糖治疗BDS的相关差异基因,对其进行功能注释和富集分析,筛选出黄精多糖治疗血虚的基因表达通路,并采用定量聚合酶链反应(quantitative polymerase chain reaction,qPCR)进行实验验证。结果与模型组比,黄精多糖(0.4 g·kg^(-1))可升高血虚小鼠红细胞、白细胞、血红蛋白、血小板、红细胞平均血红蛋白浓度等血常规指标(P<0.01),恢复小鼠体质量和体温(P<0.01或P<0.05)。转录组学分析结果显示其作用机制主要与造血细胞系、视黄醇代谢、甾类激素生物合成、血小板激活、B细胞受体信号通路、白细胞穿内皮移行等通路相关。qPCR结果表明,与血虚模型组比,黄精多糖(0.4 g·kg^(-1))可明显促进JAK1、STAT1和GATA1m RNA的表达水平(P<0.01或P<0.05)。结论黄精多糖具有治疗BDS的作用,其治疗BDS的关键差异基因主要与恢复造血功能、调节激素和免疫功能等相关。黄精多糖治疗BDS可能通过干预JAK1/STAT1信号通路发挥作用。

OBJECTIVE To explore the mechanism of steam-processed Polygonatum sibiricum polysaccharides(SPSP)in prophylaxis and treatment of mice with blood deficiency syndrome(BDS)by RNA sequencing(RNA-seq)technology.METHODS The mice were randomly divided into five groups(10 mice in each group),namely normal group,model group,SPSP groups(0.1,0.4 g·kg^(-1)),Danggui Buxue oral liquid(DOL)group.BDS model was induced in mice by acetylphenyl-hydrazine and cyclophosphamide.Blood routine,body weight and body temperature were tested after a consecutive administration for 14 d.The differential expressed genes(DEGs)related to anti-BDS by SPSP were screened through the transcriptome sequencing of the hepatic tissue in BDS mice.Functional annotation and enrichment analysis were performed to screen out the gene expression signaling pathways related to the treatment of SPSP on BDS mice.Quantitative polymerase chain reaction(qPCR)was used to verify the experiment.RESULTS Compared with the model group,SPSP(0.4 g·kg^(-1))could elevate the blood routine indexes such as red blood cell,white blood cell,hemoglobin,platelet,mean corpuscular hemoglobin concen-tration(P<0.01),and reverse the body weight and body temperature to normal(P<0.01 or P<0.05).The result of transcriptomic analysis showed that the underlying mechanism was mainly related to hematopoietic cell line,retinol metabolism,steroid hormone biosynthesis,platelet activation,B cell receptor signaling pathway,and leukocyte transendothelial migration,etc.The result of qPCR showed that SPSP(0.4 g·kg^(-1))could elevate the expression of JAK1,STAT1 and GATA1 mRNA(P<0.01 or P<0.05).CONCLUSION SPSP has therapeutic effects on BDS.The key DEGs in the treatment of BDS by SPSP are mainly related to the restoration of hematopoietic function,regulation of hormone and immune function.The mechanism of SPSP on treatment of BDS might be the regulation of JAK1/STAT1 signaling pathway.