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Outline
To investigate the regulatory effects of icariin (ICA) on the proliferation of H9C2 cardiomyocytes injured by oxygen and glucose deprivation/reperfusion (OGD/R) and on filamin B (FLNB)/β-actin (ACTB)/cyclin D1 (CCND1) signaling pathway, H9C2 cardiomyocytes were used as the research model. Cell viability was determined using the cell counting kit-8 assay. The cells were divided into three groups, normal control group, OGD/R group, and ICA group. The normal control group was cultured under normal conditions, while the other two groups were subjected to 4 hours of OGD followed by 2 hours of reperfusion. Enzyme-linked immunosorbent assay was employed to measure the levels of aspartate transaminase (AST), lactate dehydrogenase (LDH), and creatine kinase (CK). Transcriptome sequencing was performed to identify the target genes of ICA in H9C2 cells; differentially expressed genes (DEGs) were analyzed via gene ontology (GO) functional annotation and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment. Molecular docking and molecular dynamics simulation were used to evaluate the binding affinity between ICA and core targets. Compared with the OGD/R group, ICA groups at concentrations of 1-10 μmol·L-1 exerted a significant protective effect, and the levels of AST, LDH, and CK were notably decreased. Transcriptome sequencing results revealed that ICA significantly regulated 436 common DEGs in H9C2 cells after OGD/R, primarily affecting biological functions such as anatomical structure morphogenesis, protein binding, and intracellular anatomical structure, as well as signaling pathways including focal adhesion and pathways in cancer. STRING database analysis identified core targets including FLNB, ACTB, and CCND1. ICA significantly reversed the OGD/R-induced upregulation of FLNB, ACTB, and CCND1 expression. Molecular docking demonstrated strong binding between ICA and the core differential proteins, and molecular dynamics simulation confirmed that ICA significantly maintained the stability of FLNB protein. In conclusion, ICA can alleviate OGD/R-induced injury in H9C2, reduce the levels of AST, LDH, and CK, and downregulate the expression of FLNB, ACTB, and CCND1. This protective effect may be closely associated with ICA's binding to FLNB directly and subsequent mediation of the ACTB/CCND1 signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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