基于系统药理学探讨草乌改善2型糖尿病的分子机制

Exploring the Molecular Mechanism of Aconiti Kusnezoffii Radix Ameliorates Diabetes Mellitus Type 2 Based on Systems Pharmacology

关键词:  草乌  2型糖尿病  系统药理学  生物信息学  分子对接  分子机制

Key words:  Aconiti Kusnezoffii Radix  diabetes mellitus type 2  systematic pharmacology  bioinformatics  molecular docking  molecular mechanism

摘要:

目的 基于系统药理学、生物信息学、分子对接及体内外实验探讨草乌影响2型糖尿病(diabetes mellitus type 2，T2DM)的潜力及对T2DM相关症状的潜在机制。方法 利用数据库检索草乌相关化学成分、预测潜在作用靶点以及干预相关疾病；GEO数据库检索T2DM相对健康人的差异基因，与草乌作用靶点映射后置于DAVID数据库进行生物功能富集，利用单因素方差分析、二元Logistic回归分析及ROC曲线分析目标基因对于T2DM的敏感性。借助分子对接技术分析草乌化学成分与靶蛋白的结合位置及相互作用力。通过体内外T2DM模型验证草乌及其化学成分对目标蛋白表达的影响。结果 通过数据库检索与分析得到草乌干预靶点相关疾病304种(P<0.05，FDR<0.05)，选取度值最高的T2DM(Degree=59)进行分析。T2DM相对健康人差异基因与草乌潜在作用靶点取交集得到43个目标基因，进行单因素方差分析得到有显著性差异的基因共9个，二元Logistic回归分析得到5个有意义基因，ROC曲线下面积AUC>0.5的基因共3个。分子对接得到草乌化学成分异波尔定碱[(+)-Isoboldine]与蛋白APEX1、CASP1、CBFB，欧乌头碱(Napelline)与蛋白CBX1、EHMT2结合，通过氢键相互作用、疏水相互作用、可电离性和静电相互作用等不同作用力结合，从而增加配体靶向作用蛋白的能力。T2DM大鼠经草乌水提物治疗2周后，草乌可能通过改善周围神经病变进而缓解T2DM症状。并且草乌可以影响大鼠肝组织中APEX1、CASP1、CBFB、CBX1、EHMT2的蛋白表达。草乌中(+)-Isoboldine和Napelline化学成分对体外模型的目标蛋白的影响与体内实验一致。结论 本研究初步揭示草乌可作为改善T2DM周围神经病变的潜在治疗药物，为中蒙药研发以及挖掘治疗T2DM新靶标药物奠定理论基础。

Abstract:

OBJECTIVE To explore the potential of Aconiti Kusnezoffii Radix in affecting diabetes mellitus type 2(T2DM) and the potential mechanism for T2DM related symptoms based on systematic pharmacology, bioinformatics, molecular docking and in vitro and in vivo experiments. METHODS The database was used to search the related chemical components of Aconiti Kusnezoffii Radix, predict the potential targets and intervene related diseases. The differential genes of T2DM relative healthy people were retrieved from GEO database, mapped with the action target of Aconiti Kusnezoffii Radix, and placed in DAVID database for biological function enrichment. The sensitivity of the target gene to T2DM was analyzed by one-way ANOVA, binary logistic regression analysis and ROC curve. The binding position and interaction force between chemical compounds of Aconiti Kusnezoffii Radix and target proteins were analyzed by molecular docking technology. The effect of Aconiti Kusnezoffii Radix and its chemical compounds on the expression of target protein was verified by T2DM model in vivo and in vitro. RESULTS Through database retrieval and analysis, 304 kinds of target related diseases(P value<0.05, FDR<0.05) were obtained, and T2DM with the highest degree value(Degree=59) was selected and analyzed. The 43 target genes were obtained from the intersection of differential genes in T2DM relatively healthy people and potential action targets of Aconiti Kusnezoffii Radix. A total of 9 genes with significant differences were obtained by one-way ANOVA, 5 meaningful genes were obtained by binary logistic regression analysis, and 3 genes with area under ROC curve AUC>0.5 were obtained. By molecular docking (+)-Isoboldine binds to proteins APEX1, CASP1 and CBFB, Napelline binds to proteins CBX1 and EHMT2 through different forces such as hydrogen bond interaction, ligand interaction, hydrophobic interaction, ionizability and electrostatic interaction, so as to increase the ability of ligands to target proteins. After 2 weeks of treatment with Aconiti Kusnezoffii Radix aqueous extract, Aconiti Kusnezoffii Radix may alleviated the symptoms of T2DM by improving peripheral neuropathy. Moreover, Aconiti Kusnezoffii Radix could affect the protein expression of APEX1, CASP1, CBFB, CBX1 and EHMT2 in rat liver tissue. The effect of (+)-Isoboldine and Napelline chemical compounds in Aconiti Kusnezoffii Radix on the target protein of the model in vitro was consistent with that in vivo. CONCLUSION It is preliminarily revealed that Aconiti Kusnezoffii Radix can be used as a potential therapeutic drug to improve T2DM peripheral neuropathy, which lays a theoretical foundation for the research and development of Chinese Mongolian medicine and the excavation of new target drugs for the treatment of T2DM.