基于HDAC3/8分子互作技术的类叶牡丹抗类风湿性关节炎有效部位活性成分分析

目的筛选类叶牡丹Caulopyhllumrobustum抗人类风湿性关节炎(RA)有效部位中具有抑制HDAC3/8活性的成分。方法通过AutoDock计算机虚拟对接得到类叶牡丹活性成分与HDAC3/8对接结合能,再通过荧光检测方法进行结合生物活性验证。最后采用BiacoreT200分子相互作用分析系统,构建偶联HDAC受体的分子芯片,将较优活性成分与靶蛋白进行分子对接,考察成分与靶标的亲和性。结果葳严仙皂苷H、葳严仙皂苷G、葳严仙皂苷D和leonticin D与HDAC3/8均无结合能,常春藤皂苷元、刺囊酸和齐墩果酸与HDAC3/8的虚拟对接结果最好,且抑制HDAC3/8生物活性最佳。偶联HDAC受体的分子芯片后,具有结合趋势的活性成分中与靶蛋白亲和性最佳的成分是常春藤皂苷元。结论实验证明类叶牡丹抗RA有效部位中具有HDAC抑制活性的化学成分,其中与HDAC3/8结合最好的是常春藤皂苷元。

Analysis of active components of Caulopyhllum robustum against rheumatoid arthritis based on HDAC3/8 molecular interaction technology

Objective To screen for components that inhibit HDAC3/8 activity in the effective part of the Caulopyhllum robustum against rheumatoid arthritis (RA). Methods The binding energy of the active component of the C. robustum with HDAC3/8 was obtained by virtual docking with AutoDock computer, and the binding activity was verified by fluorescence detection. Finally, the Biacore T200 molecular interaction analysis system was used to construct a molecular chip coupled with HDAC receptor, and the optimal active component was molecularly docked with the target protein to investigate the affinity of the component with the target. Results Cauloside H, Cauloside G, Cauloside D, and Leonticin D have no binding energy to HDAC3/8. The virtual docking results of hederagenin, echinocystic acid and oleanolic acid and HDAC3/8 are the best, and the biological activity of HDAC3/8 is best. After coupling the molecular chip of the HDAC receptor, the component with the best affinity for the target protein in the binding tendency is hederagenin. Conclusion The experimental results showed that the active constituents of the C. robustum have the chemical component of HDAC inhibitory activity, and the best combination with HDAC3/8 is hederagenin.