Objective Exploring the mechanism of mitophagy-mediated efferocytosis in myocardial fibrosis progression based on transcriptomics and bioinformatics,with potential chinese herbal medicine prediction and experimental validation.Methods A rat model of myocardial fibrosis was constructed by injecting angiotensin Ⅱ subcutaneously.RNA extraction and high-throughput sequencing were performed on myocardial tissue of rats,and differentially expressed genes(DEGs)were detected.Efferocytosis-related genes(ERGs)were obtained from the GeneCards database,and the intersection of DEGs and ERGs was mapped using the"venneuler"package in R 4.2.0 software.Differentially expressed efferocytosis-related genes(DEERGs)were obtained,and KEGG and GO enrichment analyses were performed for DEERGs.The network analysis of protein-to-protein interactions(PPI)and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were conducted.The key genes were imported into Coremine database for chinese herbal medicine prediction.Combined with the Traditional Chinese Medicine pathogenesis of myocardial fibrosis and the previous research progress,the highest frequency of Astragali Radix,Carthami Flos and Lepidii Semen was selected for animal experiments.Masson staining was used to analyze the degree of fibrosis.Mitochondrial swelling assay and mitochondrial membrane potential detection were performed to evaluate the extent of mitochondrial dysfunction.RT-PCR and Western blot were employed to measure the mRNA and protein expression of mitophagy-related genes to assess mitophagy levels.Cardiac-resident macrophages were isolated,treated with AngⅡ,and subsequently assessed for their efferocytosis of apoptotic endothelial cells.Results There were 428 DEGs between in the control group and model group,including 165 up-regulated and 263 down-regulated DEGs.250 ERGs were identified from GeneCards database,and 12 DEERGs were obtained by drawing the intersection of DEGs and ERGs.KEGG and GO enrichment analysis of DEERGs showed that the process involved cellular burial,mitochondrial autophagy and endothelial cell regulation.PPI network analysis and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were performed.The top 10 key genes were HIF1A,PINK1,GABARAP,ARG1,PPARG,UCP2,SQSTM1,CD36,SIRT1 and RAB7A.The key genes were imported into Coremine database to search for corresponding chinese herbal medicine.The most frequent ones were Astragali Radix,Carthami Flos and Lepidii Semen.Experimental validation demonstrated that the Astragali Radix,Carthami Flos and Lepidii Semen formulation inhibited AngⅡ-induced myocardial fibrosis and improved cardiac function in rats.Notably,it significantly enhanced the efferocytosis of apoptotic endothelial cells by AngⅡ-treated resident macrophages,with the high-dose group showing the most pronounced effect(P<0.05).Mechanistic studies using the high-dose Astragali Radix,Carthami Flos and Lepidii Semen formulation group revealed that,compared to control and model groups,the treatment group exhibited reduced mitophagy and ameliorated mitochondrial dysfunction(P<0.05).Conclusion The Astragali Radix,Carthami Flos and Lepidii Semen formulation may inhibit excessive mitophagy by regulating the expression of PINK1 and P62,promote macrophage efferocytosis,and thereby suppress the formation of myocardial fibrosis.

Objective Exploring the mechanism of mitophagy-mediated efferocytosis in myocardial fibrosis progression based on transcriptomics and bioinformatics,with potential chinese herbal medicine prediction and experimental validation.Methods A rat model of myocardial fibrosis was constructed by injecting angiotensin Ⅱ subcutaneously.RNA extraction and high-throughput sequencing were performed on myocardial tissue of rats,and differentially expressed genes(DEGs)were detected.Efferocytosis-related genes(ERGs)were obtained from the GeneCards database,and the intersection of DEGs and ERGs was mapped using the"venneuler"package in R 4.2.0 software.Differentially expressed efferocytosis-related genes(DEERGs)were obtained,and KEGG and GO enrichment analyses were performed for DEERGs.The network analysis of protein-to-protein interactions(PPI)and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were conducted.The key genes were imported into Coremine database for chinese herbal medicine prediction.Combined with the Traditional Chinese Medicine pathogenesis of myocardial fibrosis and the previous research progress,the highest frequency of Astragali Radix,Carthami Flos and Lepidii Semen was selected for animal experiments.Masson staining was used to analyze the degree of fibrosis.Mitochondrial swelling assay and mitochondrial membrane potential detection were performed to evaluate the extent of mitochondrial dysfunction.RT-PCR and Western blot were employed to measure the mRNA and protein expression of mitophagy-related genes to assess mitophagy levels.Cardiac-resident macrophages were isolated,treated with AngⅡ,and subsequently assessed for their efferocytosis of apoptotic endothelial cells.Results There were 428 DEGs between in the control group and model group,including 165 up-regulated and 263 down-regulated DEGs.250 ERGs were identified from GeneCards database,and 12 DEERGs were obtained by drawing the intersection of DEGs and ERGs.KEGG and GO enrichment analysis of DEERGs showed that the process involved cellular burial,mitochondrial autophagy and endothelial cell regulation.PPI network analysis and identification of key genes of DEERGs and mitochondrial autophagy(PINK1,P62)were performed.The top 10 key genes were HIF1A,PINK1,GABARAP,ARG1,PPARG,UCP2,SQSTM1,CD36,SIRT1 and RAB7A.The key genes were imported into Coremine database to search for corresponding chinese herbal medicine.The most frequent ones were Astragali Radix,Carthami Flos and Lepidii Semen.Experimental validation demonstrated that the Astragali Radix,Carthami Flos and Lepidii Semen formulation inhibited AngⅡ-induced myocardial fibrosis and improved cardiac function in rats.Notably,it significantly enhanced the efferocytosis of apoptotic endothelial cells by AngⅡ-treated resident macrophages,with the high-dose group showing the most pronounced effect(P<0.05).Mechanistic studies using the high-dose Astragali Radix,Carthami Flos and Lepidii Semen formulation group revealed that,compared to control and model groups,the treatment group exhibited reduced mitophagy and ameliorated mitochondrial dysfunction(P<0.05).Conclusion The Astragali Radix,Carthami Flos and Lepidii Semen formulation may inhibit excessive mitophagy by regulating the expression of PINK1 and P62,promote macrophage efferocytosis,and thereby suppress the formation of myocardial fibrosis.