| 引用本文:卜祥伟1，郝晓晖1，张美珍2，王泽1，王皓朔1，史佩玉1，张润云1，倪青1，林兰1.基于高通量测序研究清润方改善2型糖尿病大鼠肝脏胰岛素抵抗的作用机制[J].世界中医药,2024,(11):. |
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| 基于高通量测序研究清润方改善2型糖尿病大鼠肝脏胰岛素抵抗的作用机制 |
| Mechanism of Qingrun Formula in Improving Hepatic Insulin Resistance of Rats with Type 2 Diabetes Mellitus Based on High-throughput Sequencing |
| 投稿时间：2023-04-21 |
| DOI：10.3969/j.issn.1673-7202.2024.11.010 |
| 中文关键词: 清润方 2型糖尿病 胰岛素抵抗 转录组测序 竞争性内源性RNA网络 差异基因 信号通路 作用机制 |
| English Keywords:Qingrun Formula Type 2 diabetes mellitus Insulin resistance Transcriptome sequencing Networks of competitive endogenous RNA Differential gene Signaling pathway Mechanism |
| 基金项目:国家自然科学基金面上项目(81973685) |
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| 中文摘要: |
| 目的：基于高通量转录组测序（RNA-seq）研究清润方改善2型糖尿病（T2DM）大鼠肝脏胰岛素抵抗的作用机制。方法：采用高脂饲料喂养联合链脲佐菌素腹腔注射构建T2DM大鼠模型，将成模大鼠采用随机数字表随机分为模型组，二甲双胍组（150 mg/kg），清润方大（11.2 g/kg）、中（5.6 g/kg）、小（2.8 g/kg）剂量组，另设正常组，灌胃干预8周。观察空腹血糖（FBG）、胰岛素抵抗指数（IRI）、胰岛素敏感指数（ISI）变化，利用RNA-seq结合生物信息学分析筛选差异表达的长链非编码RNA(lncRNA)、微RNA(miRNA)、信使RNA(mRNA)，对差异基因进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析及qPCR验证，构建竞争性内源性RNA(ceRNA)调控网络。结果：与模型组比较，干预第6、8周清润方大剂量组FBG明显降低（P<0.05）；第8周清润方各剂量组IRI降低（P<0.01），清润方大、小剂量组ISI升高（P<0.01，P<0.05）。通过差异表达筛选得到85个关键mRNA、12个miRNA、37个lncRNA，通过蛋白质-蛋白质相互作用(PPI)网络得到12个核心基因，并构建lncRNA-miRNA-mRNA网络。KEGG富集分析显示，差异基因主要涉及Janus激酶/信号转导及转录活化因子（JAK-STAT）、过氧化物酶体增殖物激活受体（PPAR）、氨基酸代谢、脂质代谢等通路。结论：清润方可能通过lncRNA-miRNA-mRNA网络调控CYP2、Acer2等基因，并影响Lpin1、Insig1等基因和PPAR、JAK-STAT、氨基酸代谢、脂质代谢等信号通路，改善2型糖尿病大鼠肝脏胰岛素抵抗。 |
| English Summary: |
| To explore the mechanism of Qingrun Formula in improving hepatic insulin resistance in rats with type 2 diabetes mellitus(T2DM) through high-throughput RNA sequencing(RNA-seq).Methods:A T2DM model of rats was established by feeding rats a high-fat diet combined with intraperitoneal injection of streptozotocin(STZ).The successfully modeled rats were randomly divided into the model group，metformin group(150 mg/kg)，and high-dose(11.2 g/kg)，medium-dose(5.6 g/kg)，and low-dose(2.8 g/kg) groups of Qingrun Formula，and a normal control group was set up.The rats were given intragastric intervention for eight weeks.Changes in fasting blood glucose(FBG)，insulin resistance index(IRI)，and insulin sensitivity index(ISI) were observed among the groups.RNA-seq technology combined with bioinformatics analysis was used to screen differentially expressed long-chain non-coding RNA(lncRNA)，micro RNA(miRNA)，and messenger RNA(mRNA).Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analyses and qPCR verification of differentially expressed genes were performed，and regulatory networks of competitive endogenous RNA(ceRNA) were constructed.Results:Compared with that in the model group，the FBG in the high-dose group of Qingrun Formula decreased significantly in the 6th and 8th week of intervention(P<0.05).In the 8th week of intervention，the IRI in all groups of Qingrun Formula decreased significantly(P<0.01)，while that in the high-dose and low-dose groups of Qingrun Formula increased significantly(P<0.01，P<0.05).85 key mRNA，12 miRNA，and 37 lncRNA were obtained through differential expression screening.12 core genes were obtained by protein-protein interaction(PPI) network，and the network of lncRNA-miRNA-mRNA was constructed.KEGG enrichment analysis revealed that differentially expressed genes were mainly involved in pathways such as JAK-STAT，PPAR，amino acid metabolism，and lipid metabolism.Conclusion:Qingrun Formula may improve hepatic insulin resistance of rats with T2DM by regulating CYP2 and Acer2 genes through the lncRNA-miRNA-mRNA network and affecting Lpin1 and Insig1 genes，as well as PPAR，JAK-STAT，amino acid metabolism，lipid metabolism，and other signaling pathways. |
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