基于反向对接法的黄连碱抗炎机制分子模拟研究

Study on anti-inflammatory mechanism of coptisine based on reverse docking method

目的:基于分子反向对接技术,以黄连碱为研究对象,对TLR4/NF-κB、p38、JAK2/STAT3 3条通路上的蛋白进行反向筛选,以筛选出黄连碱的炎症靶标蛋白。方法:获取Toll样受体4/核因子(TLR4/NF-κB)、p38促分裂原活化蛋白激酶(P38 MAPK)和Janus激酶-信号转导转录激活因子(JAK2/STAT3)3条炎症通路上的30个蛋白的晶体学结构以及黄连碱的化学结构;利用AutoDockTools对所有蛋白的晶体学结构进行标准化处理;Auto Grid对蛋白的活性位点进行格子计算;利用AutoDock对黄连碱进行反向对接模拟实验;根据对接结果自由能高低进行排序,筛选出亲和力高的靶蛋白;对筛选得到的靶蛋白进行作用力分析并作图。结果:反向筛选得到4个与黄连碱具有高亲和性的靶蛋白,4个靶蛋白分别为PI3Kδ、PI3Kγ、IKKβ、PI3Kα。结论:黄连碱对PI3Kδ、PI3Kγ、IKKβ、PI3Kα靶蛋白具有竞争抑制的活性,提示黄连碱可以通过抑制该4个靶蛋白进而阻碍炎症信号传递,影响下游蛋白的表达,发挥抗炎作用。

Objective： To investigate the mechanism of anti-inflammation of coptisine via the approach of reverse docking, which was utilized to dock coptisine with proteins in the signal pathways of TLR4/NF-κB, P38 and JAK2/STAT3. Method： 30 proteins of TLR4/NF-κB, P38 and JAK2/STAT3 were obtained from PDB website. The structure of coptisine was obtained from PubChem website. The 30 proteins were processed with standardization disposal of AutoDock tools. Then AutoGrid was utilized to calculate the grids of active sites. The docking results were ranked according to docking energy, for the purpose of selecting the proteins with high affinity for coptisine. The action force of the target protein were analyzed and plotted. Result： 4 target proteins including PI3Kδ、PI3Kγ、IKKβ and PI3Kα were found to interact with coptisine with high affinity. Conclusion： Anti-inflammation of coptisine relates to its function of competitive inhibition of the 4 target proteins, which suggests that the behavior of coptisine results in the obstruction to signal pathways. Finally the purpose of treatment for inflammation has been achieved by this method.