桑色素通过抑制MMP9表达改善慢性阻塞性肺疾病

Morin ameliorates chronic obstructive pulmonary disease by inhibiting the expression of MMP9

目的探讨桑色素通过调控基质金属蛋白酶9(MMP9)对慢性阻塞性肺疾病(COPD)动物和细胞模型的影响。方法运用SwissTargetPrediction数据库预测桑色素的基因靶点;GeneCards、OMIM及DisGenet数据库对COPD的风险基因进行预测并且借助Venny在线网站对COPD相关基因靶点和桑色素基因靶点取交集;利用String数据库完成蛋白质互作用(PPI)分析并借助Cytoscape 3.8.2软件完成拓扑学分析;借助分子对接技术对药物成分-基因靶点的对接情况进行验证;构建COPD大鼠模型并且验证桑色素对COPD大鼠模型的治疗作用;构建COPD细胞模型并且干预细胞中MMP9的表达;流式细胞术检测细胞凋亡率;ELISA检测炎性因子浓度。结果SwissTargetPrediction数据库预测得到96个桑色素的基因靶点,GeneCards、OMIM、DisGenet数据库预测得到7122个COPD的基因靶点,取交集得到67个桑色素和COPD的共同作用基因靶点;拓扑学分析显示交集基因靶点中互作用较强的前5个基因靶点为SRC、MMP9、MMP2、PTGS2、FURIN;分子对接结果显示MMP9与桑色素结合活性最好;动物实验表明桑色素能够改善COPD大鼠呼吸功能参数、肺组织病理损伤并且抑制肺组织中MMP9的表达(P<0.05);细胞实验显示桑色素能够增加COPD细胞模型的细胞活力,抑制MMP9的表达(P<0.05)。此外,桑色素能够抑制COPD细胞模型的细胞凋亡率以及炎性因子表达,但该作用被MMP9过表达载体逆转(P<0.05)。结论桑色素能够通过抑制MMP9的表达COPD细胞模型细胞凋亡以及炎性因子表达,减少动物模型肺组织病理改变。

Objective To investigate the effects of morin on chronic obstructive pulmonary disease(COPD)animal and cell model through the regulation of matrix metalloproteinase-9(MMP9).Methods SwissTargetPrediction database was used to predict the genetargets of morin,GeneCards,OMIMand DisGenet databases were used to predict the risk genes of COPD,the intersection of COPD-related gene targets and morin gene targets was obtained by using Venny online website;Protein-protein interaction(PPI)analysis was performed using the String database,and topological analysis was performed by Cytoscape 3.8.2 software.Molecular docking was used to validate the interaction between the drug components and gene targets;COPD rat models were constructed to verify the therapeutic effects of morin om COPD rat models.The COPD cell model was established to interfere with the expression of MMP9 in the cells.COPD cell models were established and the expression of MMP9 in cells was intervened;flow cytometry was used to detect cell apoptosis rate;ELISA was used to measure the concentration of inflammatory factors.Results The Swiss Target Prediction database predicted 96 gene targets of morin.GeneCards,OMIM,and DisGenet databases predicted 7122 gene targets of COPD,intersection analysis identified 67 common gene targets between morin and COPD.Topological analysis revealed that the top 5 gene targets with strongest interactions among the intersected targets were SRC,MMP9,MMP2,PTGS2,and FURIN;Molecular docking results showed the best binding activity between MMP9 and morin;animal experiments showed that morin improved respiratory function parameters and lung tissue pathological damage,and inhibited MMP9 expression in COPD rats(P<0.05);Cell experiments showed that morin increased cell viability in COPD cell models and inhibited MMP9 expression(P<0.05).In addition,morin inhibited cell apoptosis and expression of inflammatory factors in the COPD cell model,but this effect was reversed by overexpression of MMP9(P<0.05).Conclusion Morin can improve apoptosis and and expression of inflammatory cytokines in COPD cell model,reduce lung tissue pathological changes by inhibiting MMP9 expression.