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To explore the effect of berberine hydrochloride on the proliferation and differentiation of chicken myoblasts. In this study, the effects of berberine hydrochloride on cell proliferation and differentiation were detected by CCK-8 assay, EdU staining and Desmin immunofluorescence staining. The gene expression levels of myogenic factor 5 (MYF5) and myogenic differentiation 1 (MYOD) were determined by real-time quantitative PCR (RT-qPCR). The effects of berberine hydrochloride on the gene expression profiles of chicken myoblasts were also investigated by RNA sequencing analysis. The results showed that berberine hydrochloride treatment significantly promoted the proliferation of chicken myoblasts while inhibiting the expression of MYOD (P<0.01) and MYF5 (P<0.05). Desmin staining showed slightly weaker fluorescence signals in the cytoplasm of the experimental group than that of the control group, indicating that berberine hydrochloride inhibited the differentiation of chicken myoblasts to a certain extent. In addition, Gene Ontology enrichment analysis (GO) showed that the differential genes were mainly enriched in extracellular space and matrix, cell adhesion, and cell migration regulation after berberine hydrochloride treatment; Kyoto Encyclopedia of Genes and Genomes enrichment analysis (KEGG) showed that the differential genes were mainly enriched in tyrosine metabolism, TGF-β signaling pathway, MAPK signaling pathway and cell adhesion molecule-related signaling pathway. The RT-qPCR results showed that in the TGF-β pathway, the expression of Inhibitor of DNA Binding 1 (ID1) and Paired-like Homeodomain Transcription Factor 2 (PITX2) was upregulated, while Follistatin (FST) was downregulated. In the MAPK pathway, the pro-proliferative genes Insulin-like Growth Factor 2 (IGF2), Fibroblast Growth Factor 7 (FGF7), FBJ Murine Osteosarcoma Viral Oncogene Homolog (FOS), Fms-related Tyrosine Kinase 1 (FLT1), and MAPK-activated Protein Kinase 3 (MAPKAPK3) were all upregulated, whereas the calcium channel gene Calcium Voltage-gated Channel Subunit Alpha1 E (CACNA1E) was downregulated. These findings suggested that berberine hydrochloride might synergistically regulate the proliferation-differentiation balance of myoblasts through the interaction between the TGF-β and MAPK signaling pathways. In this study, we found that berberine hydrochloride promoted the proliferation of chicken myoblasts and analyzed the molecular mechanism and provided theoretical support for the efficient culture of chicken myoblasts in vitro.
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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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