TIMP3 induces gene expression partly through PI3K and their association with vascularization and heart rate

Figures

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  Figure 1. Recombinant TIMP3 (rTIMP3) modulated differential gene expression and potential pathways involved. (A) A volcano picture shows differentially expressed genes (DEGs) regulated by rTIMP3, including up-regulated (Up) and down-regulated DEGs (Down) with change fold ≥2 and p.adjust < 0.05 and other genes without significance (None). (B) KEGG pathway enrichment figure shows some pathways that were significantly regulated by rTIMP3 through DEGs in cardiomyocytes.

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  Figure 2. Some DEGs regulated by rTIMP3 were verified by real-time PCR. Neonatal rat ventricular myocytes were treated with rTIMP3 (100 ng/ml) and mRNA levels (n = 3), were determined with real-time PCR. ADAM, a disintegrin and metalloproteinase; ADAMTS7, ADAM with thrombospondin motifs 7; APLN, apelin; ATF3, activating transcription factor 3; IL, interleukin; MAP2K6, dual specificity mitogen-activated protein kinase kinase 6; MMP, matrix metalloproteinase. *p < 0.05, **p < 0.01 vs. control.

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  Figure 3. Some DEGs modulated by rTIMP3 were involved in regulation of vascularization and heart rate. DEGs regulated by rTIMP3 were predicted to increase vascularization (A) or to decrease heart rate (B) via Ingenuity Pathway Analysis (IPA) software, and their alterations were presented with heat maps illustrated by using TBtools.

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  Figure 4. Pi3k and AGTR2 participated in rTIMP3-induced gene expression. Neonatal rat ventricular myocytes were treated with rTIMP3 (100 ng/ml) for 48 h with or without pre-incubation of specific inhibitors for phosphoinositide-3-kinase (PI3K, LY294002, 10 μM) or Type-2 angiotensin II receptor (AGTR2, PD123319, 1 μM) for 30 min. Total RNAs were collected and mRNA levels of ADAM12, ADAM17, ADAMTS7, APLN, IL1β, IL6, IL33, MMP2, MMP3, and MMP9 were detected by using real-time PCR (n = 3). **p < 0.01 vs. control, # p < 0.05, ## p < 0.01, vs. rTIMP3.