结合生物信息学和分子对接技术探讨基于肝糖异生抑制策略治疗2型糖尿病的靶标蛋白标志物

Combining Bioinformatics and Molecular Docking Technology to Explore the Target Protein Markers for the Treatment of T2DM Based on Hepatic Gluconeogenesis Inhibitory Strategies

关键词:  2型糖尿病  肝糖异生  生物信息学  分子对接  蛋白标志物

Key words:  type 2 diabetes mellitus  liver gluconeogenesis  bioinformatics  molecular docking  protein marker

摘要:

目的 基于肝糖异生抑制策略筛选2型糖尿病(type 2 diabetes mellitus,T2DM)的标志性靶标蛋白。方法 从NCBI数据库下载人类肝组织基因芯片数据集,使用GEO2R在线分析工具筛选T2DM易感性基因;利用GeneCards和OMIM数据库检索糖异生相关靶点;整合数据获取T2DM-肝糖异生交互靶点,并构建蛋白质间相互作用网络筛选标志性靶标蛋白;使用Metascape数据库对靶点进行基因本体论和通路富集分析;借助分子对接预测T2DM治疗药物与标志性靶标蛋白之间的相互作用能力;最后联合二甲双胍对标志性靶标蛋白在自发性T2DM KKAy小鼠肝组织中的影响进行验证。结果 共识别出1 143个T2DM易感性基因和958个糖异生相关靶点,获得56个T2DM-肝糖异生交互靶点,并经网络分析筛选出2个标志性靶标蛋白PTPRC和VCAM1;交互靶点主要富集在葡萄糖分解代谢、典型糖酵解、葡萄糖-6-磷酸糖酵解等生物过程,通过糖酵解/糖异生、糖尿病并发症中的AGE-RAGE信号通路、内分泌抵抗等通路调节发挥作用;26个降糖药物均与标志性靶点同时具有较好的空间匹配、能量匹配以及不同程度的结合能力;T2DM模型组小鼠血糖较正常组显著增高,且在治疗后显著降低;T2DM模型组小鼠肝组织中PTPRC、VCAM1的蛋白表达水平均较正常对照组显著增高,并且均在二甲双胍治疗后显著降低。结论 本研究系统筛选并验证了肝组织中可能影响T2DM肝糖异生的2个标志性靶标蛋白,揭示了T2DM中肝糖异生调控通路,为深入研究肝糖异生防治机制,寻找新的降糖药物作用靶点提供依据。

Abstract:

OBJECTIVE To screen the marker target protein of type 2 diabetes mellitus (T2DM) based on hepatic gluconeogenesis inhibition strategy.METHODS The microarray data set of human liver tissue was downloaded from NCBI database,and T2DM susceptibility genes were screened by GEO2R online analysis tool.Gluconeogenesis related targets were retrieved from GeneCards and OMIM databases.The interaction targets between T2DM and hepatic gluconeogenesis were obtained by integrating data,and the marker target proteins were screened by constructing a protein-protein interaction network.Gene ontology and pathway enrichment analysis were performed in Metascape database.The interaction ability between T2DM therapeutic drugs and marker target proteins was predicted by means of molecular docking.Finally,the effect of the marker target proteins in the liver tissue of spontaneous T2DM KKAy mice was validated by combining metformin.RESULTS A total of 1 143 T2DM susceptibility genes and 958 gluconeogenesis related targets were identified,56 targets of T2DM interacting with hepatic gluconeogenesis were obtained,and two marker target proteins PTPRC and VCAM1 were screened by network analysis.The interaction targets were mainly enriched in glucose catabolic process,canonical glycolysis,glycolytic process through glucose-6-phosphate,and other biological processes.And they functioned through the regulation of pathways such as Glycolysis/Gluconeogenesis,AGE-RAGE signaling pathway in diabetic complications,and endocrine resistance.The 26 hypoglycemic drugs all had good spatial matching,energy matching and different degrees of binding ability with marker targets.The blood glucose of T2DM model group was significantly higher than that of normal group,and decreased significantly after treatment.The protein expression levels of PTPRC and VCAM1 in the liver tissue of the mice in the T2DM model group were significantly higher than those in the normal group,and they were significantly lower after metformin treatment.CONCLUSION This study systematically screened and verified two marker target proteins in human liver tissue that may affect hepatic gluconeogenesis in T2DM,reveal the regulatory pathway of hepatic gluconeogenesis in T2DM,and provide a basis for in-depth study of the prevention and treatment mechanism of hepatic gluconeogenesis and looking for new targets of hypoglycemic drugs.