Myc is involved in Genistein protecting against LPS-induced myocarditis in vitro through mediating MAPK/JNK signaling pathway

Figures

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  Figure 1. Genistein alleviated LPS-induced H9c2 cells damage by limiting apoptosis and the expression of inflammatory factors (A,B) The results from CCK-8 and MTT assays showed that the decreased proliferation of H9c2 cells induced by LPS was attenuated with the help of Genistein. (C,D) The increased apoptosis of H9c2 cells caused by LPS was relieved after Genistein treatment. The up-regulation of pro-inflammatory factors including TNF-α (E), IL-6 (F), and IL-8 (G) in H9c2 cells caused by LPS were inhibited under the guidance of Genistein. (H) The down-regulation of anti-inflammatory factor IL-10 in H9c2 cells caused by LPS was suppressed with the support of Genistein. **P<0.01 vs. control, ##P<0.01 vs. LPS.

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  Figure 2. Searching for genes and signaling pathway related to Genistein with the guidance of several bioinformatics prediction websites (A,B) The 104 target proteins were obtained with the help of String database. (C) The top 10 genes were acquired by sorting the 104 genes encoding these proteins with the support of hubgene analysis. (D) Myc gene was acquired by the intersection with the target genes and differential genes. (E) The expression of Myc in myocarditis was up-regulated by inquiring the GEO database, P=0.0022. (F) KEGG pathway enrichment analysis was performed to explore the potential signaling pathway related to Myc.

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  Figure 3. Associations among Genistein, Myc and MAPK pathway were identified by Western blot (A,B) The expression of Myc, p-P38 MAPK, P38 MAPK, p-JNK, and JNK were detected with the help of Western blot after different treatment. **P<0.01 vs. control, ##P<0.01 vs. LPS.

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  Figure 4. The protective effects of Genistein on H9c2 cells were mediated by Myc (A) The interference efficiency of Myc was proved to be successful. The H9c2 cells activity was evaluated by CCK-8 (B) and MTT kits (C) after different treatment. (D,E) The apoptosis of H9c2 cells was assessed by flow cytometry after different treatment. (F–I) The inflammatory factors were calculated by ELISA kit after different stimulation. **P<0.01 vs. LPS, #P<0.05 vs. LPS+Genistein, &P<0.05 vs. LPS+si-Myc.

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  Figure 5. The mitigation effects of Genistein on H9c2 cells were partly regulated by MAPK/JNK pathway (A,B) The expression of Myc, p-P38 MAPK, P38 MAPK, p-JNK, and JNK were estimated by Western blot after different stimulation. **P<0.01 vs. LPS, ##P<0.01 vs. LPS+Genistein, &&P<0.01 vs. LPS+si-Myc.