基于转录组和蛋白质组学及实验验证探究通脑饮减轻大鼠脑缺血再灌注损伤的机制OA

Objective To investigate the molecular targets and mechanism of Tongnao Decoction(TND)in alleviating cerebral ischemia-reperfusion injury(CIRI).Methods A total of eighteen Sprague-Dawley(SD)rats were randomly divided into a control group(sham operation group),a model group and a TND group,with 6 rats in each group.The middle cerebral artery occlusion(MCAO)model was used to simulate CIRI in the model group and the TND group.After the successful modeling,the TND group received water extract of TND by daily intragastric administration at a dose of 10 g/(kg·d),while the control group and the model group were gavaged with sterile distilled water.All groups received the administration volume at 1 mL/100 g body weight for 1 week.Neurological deficit severity was evaluated by Zea Longa score;cerebral infarct volume was observed by 2,3,5-triphenyltetrazolium chloride(TTC)staining;histopathological morphology of brain tissue was observed by hematoxylin-eosin(HE)staining to assess the neuroprotective effect.Transcriptomics and proteomics were combined to screen differentially expressed genes/proteins and overlapping targets between control group vs.model group and model group vs.TND group,followed by functional enrichment analysis.Finally,key targets were verified by qRT-PCR and Western blot.Results Compared with the control group,the model group showed a significantly higher Zea Longa score(P<0.01),and a larger percentage of infarct area(P<0.01).Compared with the model group,the Zea Longa score was significantly decreased(P<0.01)and a smaller percentage of infarct area was observed(P<0.01)in the TND group.Integrated transcriptomic and proteomic analysis revealed that TND significantly modulated the complement and coagulation cascades pathway and the ECM-receptor interaction pathway.Molecular biological experiments confirmed that TND upregulated the genes of SPP1,Fga,Serpinf2,F2,A2m,Itga5,Kng2,C4b,and Usp18,as well as the protein expressions of SPP1 and Serpinf2.Its mechanism may be related to the regulation of complement and coagulation cascade and ECM-receptor interaction pathway.Conclusion Tongnao Decoction may exert alleviation effect on CIRI by regulating complement and coagulation cascade and ECM-receptor interaction.