基于转录组测序、网络药理学探究益气活血方促进心肌梗死大鼠血管新生的机制OA

Objective To investigate the regulatory mechanism of Yiqi Huoxue Formula(YQHXF)on angiogenesis in myocardial infarction(MI)rats through transcriptome sequencing and network pharmacology.Methods An MI model was established via left anterior descending coronary artery ligation in 12 male SD rats.After successful modeling,the rats were divided into model and YQHXF groups using a random number table method,with six rats per group.The remaining six rats in the sham operation group were only threaded without undergoing ligation.On the second day after the operation,the YQHXF group received intragastric administration of YQHXF(8.2 g/kg),whereas the other two groups received an equal volume of distilled water,once a day for 28 days.Cardiac function was assessed using echocardiography to measure left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-systolic volume(LVESV),and left ventricular end-diastolic volume(LVEDV).The appearance of the heart was observed and photographed.Inflammatory cell infiltration and collagen fiber proliferation in the MI border area were observed using hematoxylin-eosin(HE)staining and Masson staining.Immunofluorescence staining was used to detect the fluorescence expression of CD31 in the MI border area.Transcriptome sequencing was used to identify differentially expressed genes(DEGs)between the sham operation and model groups,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed.Network pharmacology was used to screen the targets of YQHXF.These DEGs were then intersected with targets of YQHXF,and GO and KEGG analyses were performed to predict core targets and pathways involved in YQHXF-mediated post-MI angiogenesis.Results Hearts in the sham operation group appeared reddish and smooth with no significant necrosis,whereas those in the model group were pale,wrinkled,and exhibited apparent MI in the ligation area and surrounding regions.Hearts in the YQHXF group also showed whitish myocardial tissue but had a reduced MI size compared to the model group,along with significant cardiac enlargement.Compared to the sham operation group,the model group had significantly decreased LVEF and LVFS,and increased LVESV and LVEDV(P＜0.05).Compared to the model group,the YQHXF group showed increased LVEF and LVFS,and decreased LVESV and LVEDV(P＜0.05).HE staining showed neatly arranged and intact cardiomyocytes in the sham operation group.In contrast,the model group exhibited irregular cardiomyocyte morphology and arrangement with extensive inflammatory cell infiltration in the MI border area.The YQHXF group showed reduced inflammatory cell infiltration compared to the model group.Masson staining revealed red-stained,well-arranged,and clear myocardial fibers in the sham operation group.The model group had blurred myocardial cell boundaries and extensive blue collagen fiber proliferation.The YQHXF group showed reduced blue collagen fiber and a more orderly myocardial cell arrangement compared to the model group.Compared with the sham operation group,the CD31 fluorescence intensity in the model group was increased(P＜0.05).Compared with the model group,the CD31 fluorescence intensity was increased in the YQHXF group(P＜0.05).Transcriptome sequencing identified 691 DEGs between the model and sham operation groups.GO analysis involved cell biological processes,including extracellular matrix(ECM)and extracellular space structure,and KEGG enrichment was primarily in pathways such as ECM-receptor interaction and transforming growth factor-β signaling pathways.Network pharmacology retrieval identified 47 main components of YQHXF and 756 potential targets.Intersecting these with the DEGs of transcriptome sequencing,a total of 31 core targets for YQHXF in treating post-MI angiogenesis were obtained.GO and KEGG analyses predicted that YQHXF may regulate angiogenesis through calcium ion transport and the vascular endothelial growth factor(VEGF)pathway.Conclusion YQHXF may promote angiogenesis after MI through calcium ion transport and the VEGF pathway,thereby improving cardiac structure and function.