基于网络药理学和实验验证探讨大黄素改善脓毒症急性肺损伤的分子机制OA

Objective To investigate the molecular mechanism of emodin in improving sepsis-induced acute lung injury using network pharmacology and experimental validation.Methods The targets of emodin were predicted using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SwissTargetPrediction,and Similarity Ensemble Approach(SEA)database.Sepsis-related targets were obtained from DrugBank,Therapeutic Target Database(TTD),and Disease Gene Network(DisGeNET)database.The intersection of emodin targets and sepsis targets was identified.Biological function and pathway enrichment analyses of the common targets were performed using Functional Enrichment Analysis Tool(FunRich).The"component-target-pathway-disease"network of emodin was constructed using Cytoscape software,and molecular docking of emodin with the common targets was performed using the DockThor molecular docking platform.Thirty Sprague-Dawley(SD)rats were randomly divided into a sham operation group,a cecal ligation and puncture(CLP)group,and an emodin group,with 10 rats in each group.The sepsis model was induced using CLP.The sham operation group was subjected to laparotomy without ligation or puncture.The emodin group received emodin(35 mg·kg-1·d-1)by gavage for 5 consecutive days before modeling.The overall status of rats was evaluated by survival rate,motor activity,and body temperature changes.Lung injury was assessed by hematoxylin-eosin(HE)staining and lung wet/dry(W/D)weight ratio.The mRNA expression level of caspase-3 was detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction(RT-qPCR),and the protein expression level of cleaved caspase-3 was measured by Western blotting.Results Network analysis showed that there were 98 emodin-related targets,156 sepsis-related targets,and 10 common targets.These common targets were involved in biological processes such as apoptosis,protein metabolism,and energy metabolism,and were enriched in signaling pathways including adenosine diphosphate ribosylation factor 6(Arf6),phosphatidylinositol 3-kinase(PI3K),and focal adhesion kinase(FAK).Molecular docking demonstrated that emodin had good binding affinity with the 10 common targets,including caspase-3,coagulation factor Ⅶ(F7),and matrix metalloproteinase-9(MMP-9).In animal experiments,compared with the sham operation group,the CLP group showed significantly decreased survival rate[50%(5/10)vs.100%(10/10)],with manifestations including persistent fever,chills,decreased activity,reduced food intake,and decreased urine output.Pathological lung injury,including alveolar structure disruption and inflammatory cell infiltration,was observed.The lung W/D ratio,caspase-3 mRNA expression,and cleaved caspase-3 protein expression were significantly increased[lung W/D ratio:6.33±0.25 vs.4.97±0.25,caspase-3 mRNA expression(2-ΔΔCt):65.61±14.55 vs.1.00±0.02,cleaved caspase-3 protein expression(cleaved caspase-3/β-actin):1.92±0.17 vs.1.00±0.01,all P<0.01].Compared with the CLP group,the emodin group showed significantly increased survival rate[60%(6/10)vs.50%(5/10)],and significantly decreased lung W/D ratio,caspase-3 mRNA expression,and cleaved caspase-3 protein expression[lung W/D ratio:6.03±0.21 vs.6.33±0.25,caspase-3 mRNA expression(2-ΔΔCt):30.11±8.57 vs.65.61±14.55,cleaved caspase-3 protein expression(cleaved caspase-3/β-actin):1.36±0.06 vs.1.92±0.17,all P<0.05].Conclusions This study explored the potential mechanism of emodin in improving sepsis-induced acute lung injury through network pharmacology,and verified the protective effect of emodin on sepsis-induced lung injury and its regulatory effect on the caspase-3 pathway,which providesa scientific basis for the further application of emodin in the prevention and treatment of sepsis.