1. bookVolume 16 (2022): Edition 2 (April 2022)
Détails du magazine
License
Format
Magazine
eISSN
1875-855X
Première parution
01 Jun 2007
Périodicité
6 fois par an
Langues
Anglais
Accès libre

Serotonin receptor subtype-2B signaling is associated with interleukin-18-induced cardiomyoblast hypertrophy in vitro

Publié en ligne: 29 Apr 2022
Volume & Edition: Volume 16 (2022) - Edition 2 (April 2022)
Pages: 79 - 87
Détails du magazine
License
Format
Magazine
eISSN
1875-855X
Première parution
01 Jun 2007
Périodicité
6 fois par an
Langues
Anglais
Abstract Background

In patients with heart failure, interleukin-18 (IL-18) levels increase in the circulatory system and injured myocardial tissue. Serotonin (5-hydroxytryptamine) receptors subtype 2B (HTR2B) play an essential role in cardiac function and development, and their overexpression in rats leads to myocardial hypertrophy. Epigallocatechin gallate (EGCG) is cardioprotective in myocardial ischemia–reperfusion injury in rats and can prevent pressure overload-mediated cardiac hypertrophy in vivo. Mice deficient in peroxisome proliferator-activated receptor delta (PPARδ) can have cardiac dysfunction, myocardial hypertrophy, and heart failure. Matrix metalloproteinases (MMPs) are possibly involved in cardiac remodeling. However, the relationship between IL-18 signaling, cardiac hypertrophy, and the molecular mechanisms involved remain to be fully elucidated.

Objectives

To elucidate the relationship between HTR2B and IL-18-induced myocardial hypertrophy and examine the antihypertrophic effects of EGCG and PPARδ.

Methods

We induced H9c2 cardiomyoblast hypertrophy with IL-18 in vitro and investigated the downstream signaling by real-time polymerase chain reaction (PCR) and western blotting. Hypertrophy was assessed by flow cytometry. We determined the effects of EGCG and PPARδ on IL-18-induced hypertrophic signaling via HTR2B-dependent mechanisms.

Results

IL-18-induced H9c2 hypertrophy upregulated brain natriuretic peptide (BNP) protein and mRNA expression by inducing the expression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and the hypertrophy was attenuated by pretreatment with EGCG (20 μM) and L-165,041 (2 μM), a PPARδ agonist. IL-18 upregulated the expression of HTR2B, which was inhibited by pretreatment with EGCG and L-165,041. SB215505 (0.1 μM), a HTR2B antagonist and siRNA for HTR2B, attenuated H9c2 hypertrophy significantly. Inhibition of HTR2B also downregulated the expression of MMP-3 and MMP-9.

Conclusions

IL-18 and HTR2B play critical roles in cardiomyoblast hypertrophy. EGCG and L-165,041 inhibit the expression of HTR2B and augment remodeling of H9c2 cardiomyoblasts, possibly mediated by MMP-3 and MMP-9.

Keywords

Heart hypertrophy can cause cardiac failure and even death; consequently, cardiac remodeling is an important clinical problem [1]. An obvious change in morphology is a key characteristic of cardiac hypertrophy; the change is due mainly to cardiomyocyte enlargement, which causes the heart wall to thicken, and thus reduces the size of the ventricular chambers [2].

Interleukin-18 (IL-18)-induced ventricular cell hypertrophy induces the expression of atrial natriuretic peptide (ANP) mRNA and protein synthesis. IL-18 also induces the expression of GATA4 transcription factor in vitro, and activates protein kinase B (Akt) via a phosphatidylinositol 3-kinase (PI3K)–phosphoinositide-dependent kinase (PDK1) pathway [3, 4]. Activation of these pathways by IL-18 signaling may be related to cardiac hypertrophy. Clinically, higher serum IL-18 levels are associated with the occurrence of congestive heart failure, coronary artery disease, and myocardial ischemia, and IL-18 is found in symptomatic (unstable) atherosclerotic plaques [5, 6]. This suggests that chronic serum IL-18 elevation is associated with cardiac hypertrophy. Therefore, IL-18 is considered to play an important role in maladaptive myocardial hypertrophy [7]. The relationship between IL-18 signaling and cardiac hypertrophy remains to be fully elucidated. Here, we sought to investigate possible mechanisms of cardiomyocyte hypertrophy induced by IL-18.

Serotonin (5-hydroxytryptamine or 5-HT) receptor 2B (HTR2B) is a subtype of 5-HT receptor. HTR2B are expressed mainly in the embryonic development of mouse, rat, and adult human cardiovascular tissue [8]. 5-HT plays an important role in regulating heart development and function through HTR2B [9]. In the hearts of rats, overexpression of HTR2B induces mitochondrial hyperplasia, leading to myocardial hypertrophy [10]. 5-HT-induced hypertrophy is observed in adult rat left ventricular cell cardiomyocytes in vitro. After mechanical stress, increased HTR2B expression in cardiomyocytes leads to increased expression of brain natriuretic peptide (BNP), which is regulated through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) translocation [11]. Myocardial hypertrophy induced by isoproterenol and HTR2B plays a role in myocardial fibroblast signaling [12]. It seems clear that HTR2B plays an essential role in cardiac function and development. However, like the relationship between IL-18 signaling and cardiac hypertrophy, the role of HTR2B in cardiomyocyte hypertrophy remains to be fully elucidated. Manipulating HTR2B may provide a new approach to treatment of myocardial hypertrophy or heart failure.

Peroxisome proliferator-activated receptors (PPARs) are involved in the regulation of lipid metabolism and glucose homeostasis. In the cardiovascular system, they are involved in the regulation of cell growth. Mice deficient in PPARδ have been found to have cardiac dysfunction, myocardial hypertrophy, and heart failure, evidence that PPARδ plays a role in myocardial pathology [13]. Epigallocatechin gallate (EGCG), a polyphenol catechin component of green tea, can prevent the hypertrophy of vascular smooth muscle cells induced by angiotensin II by inhibiting c-Jun N-terminal kinases [14, 15]. These findings suggest that EGCG could regulate myocardial hypertrophy. HTR2 and monoamine oxidase A (MAO-A) contribute to H9c2 cardiomyoblast hypertrophy, and the MAO-A-dependent hypertrophic response required activation of extracellular signal-regulated kinases (ERKs) [16].

Here, we examined the effect of EGCG and a PPARδ agonist (L-165,041) on IL-18-induced H9c2 hypertrophy. In addition, we sought to elucidate the relationship between IL-18 and HTR2B-induced cardiomyoblast hypertrophy.

Methods
Cell culture and reagents

H9c2 myoblasts were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan), and the detection and verification of the cardiomyoblast cell line was based on Kimes [17]. Cells were cultured in Dulbecco's modified Eagle's medium (Gibco) containing 10% fetal bovine serum (Gibco) and 1% antibiotic–antimycotic (Gibco), and were maintained in an incubator at 37 °C under a humidified atmosphere of 5% CO2 in air. Cardiomyoblasts in the experimental groups were pretreated with serotonin at 10 μM, EGCG (Sigma-Aldrich) at 20 μM, PPARδ agonist L-165,041 (Sigma-Aldrich) at 2 μM, or HTR2B antagonist SB215505 (Sigma-Aldrich) at 0.1 μM for 30 min, and were subsequently treated with IL-18 (ProSpec; Ness-Ziona, Israel) at 0.3 μg/mL for 18 h.

Western blotting

Nuclear or total protein was extracted from cardiomyoblasts homogenized in cell lysis buffer (Pro-Prep; iNtRON Biotechnology; Gyeonggi-do, Korea). Protein concentrations were determined using a BCA Assay Kit (Thermo Fisher Scientific). We mixed 30 μg protein samples with sample buffer (150 μM NaCl, 1% Triton X-100), 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate (SDS), heated the mixture in a boiling water bath for 5 min, and separated the proteins by SDS–10% polyacrylamide gel electrophoresis (PAGE) under denaturing conditions (gel 10 cm × 8 cm × 1 mm; running voltage 80 V for 2 h; Hoefer SE260 Protein Electrophoresis Unit). Proteins in the gel were transferred (running voltage: 80 mA for 80 min; Hoefer TE70XP Semi-Dry Transfer) to PVDF membranes (Immobilon-P transfer membrane; Millipore), which were then soaked in methanol for 5 min. Nonspecific binding sites on the membranes were blocked with 5% skim milk in phosphate-buffered saline (PBS) containing 0.1% Tween 20 (Tween) for 60 min, and then incubated with anti-NIK (1:5,000 in PBS-Tween, Santa Cruz Biotechnology cat. No. sc-8417; Research Resource Identifier (RRID): AB_628021, https://scicrunch.org/resources), anti-NF-κB (1:5000, Santa Cruz Biotechnology cat. No. sc-8008; RRID: AB_628017), anti-BNP (1:5000, Santa Cruz Biotechnology cat. No. sc-271185; RRID: AB_10609757), anti-MMP-3 (1:5000, Santa Cruz Biotechnology cat. No. sc-21732; RRID: AB_627958), anti-MMP-9 (1:5000, Santa Cruz Biotechnology cat. No. sc-21733; RRID: AB_627959), anti-HTR2B (1:5000, Santa Cruz Biotechnology cat. No. sc-25647; RRID: AB_2124523), anti-C23 (1:5000, Santa Cruz Biotechnology cat. No. sc-8031; RRID: AB_670271), or anti-β-actin (1:5000, Santa Cruz Biotechnology cat. No. sc-47778; RRID: AB_626632) antibodies overnight at 4 °C. The membranes were washed with PBS-Tween and subsequently incubated with horseradish peroxidase-conjugated secondary antibody (anti-goat, 1:5000 in PBS-Tween, Santa Cruz Biotechnology cat. No. sc-2020; RRID: AB_631728; anti-rabbit, 1:5000, Santa Cruz Biotechnology cat. No. sc-2004; RRID: AB_631746, or anti-mouse, 1:5000, Santa Cruz Biotechnology cat. No. sc-2005, RRID: AB_631736) as appropriate for 1 h at room temperature. Signals were visualized by enhanced chemiluminescent detection.

Real-time quantitative polymerase chain reaction (qPCR)

Total RNA was extracted from H9c2 using TRIzol and reverse-transcribed to cDNA. Amplification reactions were performed in volumes of 50 μL containing reaction buffer, 25 μL Smart Quant Green Master Mix with dUTP (Protech Technology Enterprise; Taipei, Taiwan), 4 mM MgCl2, 50 ng template cDNA, and 1 μM target primer. Primer sequences were as follows: BNP (forward: 5′-AGGCAGAGTCAGAAGCCAGAGT-3′; reverse: 5′-CTTAGGTCTCAAGACAGCGCCT-3′), HTR2B (forward: 5′-GCCTTCTTCACACCTCTTGC-3′; reverse: 5′-GTCCTTTCGAGAACCATCCA-3′), or β-actin (forward: 5′-ATGCCAACACAGTGCTGTCTGG-3′; reverse: 5′-TACTCCTGCTTGCTGATCCACAT-3′). The amplification was detected using a StepOne Real-Time PCR System (Thermo Fisher Scientific). The initial denaturizing phase was 10 min at 95 °C followed by an amplification phase as described following: denaturation at 95 °C for 15 s; annealing and elongation at 60 °C for 60 s, 40 cycles; the final elongation step was 95 °C for 15 s, 60 °C for 60 s, and 95 °C for 15 s. The BNP and HTR2B Ct values were obtained and the relative fold change in gene expression was calculated as fold changes in gene expression after normalizing to β-actin using the formula 2−ΔΔCt.

Cardiomyoblast hypertrophy analysis

H9c2 were seeded (1 × 106 cells) in 6 cm diameter dishes. The cells were pretreated with EGCG, L-164,041, or SB215505 for 30 min, and subsequently treated with IL-18 for 18 h. After treatment, the cells were harvested, washed, and resuspended in PBS. Cells were maintained on ice for 30 min. At least 104 cells were collected and analyzed using a CytoFLEX LX Flow Cytometer (Beckman Coulter). The size of cells was determined using CytExpert version 2.2 (Beckman Coulter).

RNA interference

For HTR2B siRNA (Dharmacon) transfection, H9c2 were seeded (1 × 106 cells) in 6 cm diameter dishes before transfection. H9c2 were transfected with 2 μg HTR2B siRNA using Lipocurax siRNA transfection reagents (Ambo Life; Taoyuan, Taiwan), and then cultured in an incubator at 37 °C under a humidified atmosphere of 5% CO2 in air for 18 h. The transfected cells were treated with IL-18 for 18 h. Protein samples were analyzed by western blotting.

Statistical analysis

All data were analyzed using IBM SPSS Statistics for Windows (version 24). The data are normalized and expressed as mean ± standard error of mean (SEM) and individual data. Data between 2 groups were compared using a Mann–Whiney U test, and a Kruskal–Wallis test was used to compare data from 3 or more independent groups. Differences between group data with P < 0.05 were considered significant.

Results
EGCG and PPARδ agonist (L-165,041) attenuated IL-18-induced inflammation in H9c2 cardiomyoblasts

H9c2 were incubated with 0.3 μg/mL IL-18 for 18 h. IL-18 significantly increased the expression of NF-κB inducing kinase (NIK) in the H9c2 (Figure 1A). Pretreatment of H9c2 with EGCG or L-165,041 significantly attenuated the NIK expression otherwise significantly upregulated by incubation with IL-18 for 18 h. Both EGCG and L-165,041 significantly attenuated the nuclear NF-κB (nuNF-κB) expression otherwise significantly upregulated by incubation with IL-18 after 18 h (Figure 1B).

Figure 1

Pretreatment with 20 mM ECGC and 2 mM L-165,041 attenuated IL-18-induced inflammatory markers in H9c2 cardiomyoblasts. (A) NIK production significantly was inhibited by EGCG or L-165,041 (n = 4). (B) EGCG and L-165,041 significantly attenuated IL-18-induced nuNF-κB upregulation (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18 alone. ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; NIK, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inducing kinase; nuNF-κB, nuclear NF-κB; SEM: standard error of mean.

IL-18 upregulates BNP mRNA and protein in H9c2 cardiomyoblasts

Real-time qPCR showed that incubation with IL-18 for 18 h significantly upregulated BNP mRNA (P < 0.01) and pretreatment with EGCG or L-165,041 significantly attenuated the expression of BNP otherwise significantly upregulated by incubation with IL-18 (Figure 2A). Western blotting showed that the significant upregulation of BNP after incubation with IL-18 for 18 h was attenuated by pretreatment with EGCG or L-165,041 (P < 0.01 and P < 0.05, respectively; Figure 2B).

Figure 2

Effects of 20 mM ECGC and 2 mM L-165,041 treatment on IL-18-induced BNP mRNA and protein expressions in H9c2 cardiomyoblasts. Treatment with IL-18 significantly increased the BNP (n = 4) (A) mRNA and (B) protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. BNP, brain natriuretic peptide; ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SEM: standard error of mean.

IL-18 upregulates HTR2B mRNA and protein in H9c2 cardiomyoblasts

H9c2 were incubated with 0.3 μg/mL IL-18 for various times. IL-18 significantly increased the expression of HTR2B protein in H9c2 after 12 h and 18 h (P = 0.011 and P = 0.007, respectively), but the expression had returned to control levels within 24 h (Figure 3A). Serotonin alone significantly increased HTR2B mRNA expression (P = 0.03). Both EGCG and L-165,041 significantly attenuated the HTR2B mRNA upregulation by IL-18 after 18 h (P = 0.012 and P = 0.025, respectively). However, after pretreatment with SB215505, the attenuation effect was diminished (Figure 3B). Consistent with this finding, HTR2B protein was significantly upregulated by incubation with IL-18 for 18 h, as found by western blotting (P < 0.01). Pretreatment with EGCG and L-165,041 significantly attenuated the increased HTR2B protein expression seen after incubation with IL-18 (Figure 3C).

Figure 3

Pretreatment with 20 μM EGCG and 2 μM L-165,041 inhibited IL-18-induced HTR2B upregulation. (A) H9c2 cardiomyoblasts treated with IL-18 for up to 24 h, IL-18-induced HTR2B protein expression. Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 4). (B) Serotonin (S) alone (10 μM) significantly increased the expression of HTR2B mRNA. H9c2 were treated IL-18 for 18 h. Pretreatment with EGCG and L-165,041 inhibited IL-18-induced HTR2B mRNA upregulation (n = 5) and (C) HTR2B protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared to IL-18 treatment alone. HTR2B, 5-hydroxytryptamine receptor 2B; ECGC (E), epigallocatechin gallate; IL-18, Interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARδ) agonist; SB215505 (SB), HTR2B antagonist; serotonin, 5-hydroxytryptamine.

Relationship between IL-18-induced H9c2 cardiomyoblast hypertrophy and HTR2B

Incubation of H9c2 with IL-18 increased the size of the cardiomyoblasts after 18 h (P = 0.031). Pretreatment with HTR2B antagonist (SB215505) significantly inhibited IL-18-induced cell hypertrophy (P = 0.031). EGCG and L-165,041 did not augment the effect of SB215505 (Figure 4). siRNA alone acted as a negative control (P < 0.01; Figure 5A). Incubation of H9c2 with IL-18 for 18 h upregulated HTR2B protein expression (P < 0.01), and siHTR2B treatment significantly inhibited the expression of HTR2B (P = 0.042; Figure 5B).

Figure 4

Pretreatment with 0.1 mM SB215505, 20 mM EGCG, and 2 mM L-165,041 inhibited IL-18-induced H9c2 hypertrophy. H9c2 hypertrophy was inhibited by SB215505, EGCG, and L-165,041, as analyzed by flow cytometry. FSC reflects the relative size of cells. The relative average size for control was 282; IL-18 was 354; IL-18 + SB was 304; IL-18 + SB + EGCG was 272; and IL-18 + SB + L-165,041 was 273 (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. ECGC (E), epigallocatechin gallate; FSC, forward scatter; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SB215505 (SB), HTR2B antagonist; SEM, standard error of mean; SSC, side scatter. HTR2B, 5-hydroxytryptamine receptor 2B.

Figure 5

siHTR2B significantly attenuated IL-18-induced HTR2B expression. The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). (A) Pretreatment with siHTR2B significantly decreased HTR2B mRNA expression (n = 5). Differences from control were assessed using a Mann–Whitney U test. (B) Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 5). Pretreatment with siHTR2B significantly inhibited IL-18-induced HTR2B protein expression. Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; SEM, standard error of mean.

IL-18 induces H9c2 cardiomyoblast hypertrophy through the matrix metalloproteinase-3 (MMP-3) and MMP-9

Incubation of H9c2 with IL-18 for 18 h significantly upregulated their expression of MMP-3 protein (P < 0.01). Pretreatment with siHTR2B attenuated the IL-18-induced MMP-3 protein expression (P = 0.034; Figure 6). Incubation of H9c2 with IL-18 for 18 h significantly upregulated the expression of MMP-9 protein (P = 0.012), and pretreatment with siHTR2B significantly attenuated the IL-18-induced MMP-9 protein expression (P = 0.004; Figure 6).

Figure 6

Pretreatment with siHTR2B significantly attenuated IL-18-induced MMP-3 and MMP-9 expression. Total protein was extracted and analyzed by western blotting for MMP-3 and MMP-9 expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines; dashed, MMP-3; solid, MMP-9) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18 alone. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; MMP-3, matrix metalloproteinase-3; MMP-9, matrix metalloproteinase-9; SEM, standard error of mean.

Discussion

The present results confirm that the mechanism of IL- 18- induced H9c2 cardiomyoblast hypertrophy involves regulation of NIK, NF-κB, and BNP pathways, and the expression of HTR2B. HTR2B are considered to play a crucial role and chronic activation of HTR2B might contribute to cardiomyocyte hypertrophy [18], which results from its downstream pathways. HTR2B are considered to play an integral role in IL-18-induced H9c2 hypertrophy and remodeling of myocardial cells. To determine the effect of the PPARδ agonist, we chose H9c2, which have high PPAR expression [19]. However, further experiments will need to be conducted using primary cardiomyocyte cultures and in vivo to confirm our findings.

There are various causes of cardiac hypertrophy, and one of them is IL-18 acting as a cellular prehypertrophic cytokine. In previous studies, IL-18 was found to act via PI3K/PDK1/Akt/GATA4-induced cardiac hypertrophy and play an important role in cardiac remodeling and heart failure. IL-18 also induced ANP gene transcription to cause hypertrophy [20]. IL-18-induced human cardiac microvascular endothelial cell death via a NF-κB-dependent signaling pathway [21]. Inhibition of NF-κB in vivo was sufficient to attenuate angiotensin II- and isoproterenol-induced hypertrophy of cardiomyocytes [22]. Together these results indicate that NF-κB plays an important role in cardiac hypertrophy.

In the present study, western blotting showed that IL-18 upregulated NIK and nuNF-κB in an inflammatory NIK/NF-κB signaling pathway in H9c2. In previous studies of cultured cardiomyocytes, the increase in the size of cardiomyocytes subjected to a hypertrophic stimulus was associated with increased expression of natriuretic peptides at both the mRNA and protein levels [23]. Previous studies have found that IL-18 induces expression of NF-κB [24], which impairs cardiac function, and was dependent on myeloid differentiation 88 (MyD88) → interleukin 1 receptor (IL-1R) associated kinase (IRAK) → TNF receptor-associated factor 6 (TRAF6) → PI3K → Akt → induced IκB kinase (IKK) → NF-κB inducing the expression of fibronectin [3, 4, 21], and further mediated myocardial hypertrophy. In the present study, we showed BNP mRNA and protein expression were significantly upregulated after IL-18 stimulation of H9c2 cardiomyoblasts. IL-18 caused NF-κB-induced hypertrophy of H9c2 and increased BNP expression. We observed the expression of NIK, NF-κB, and BNP individually, and found an increasing trend after 18 h of IL-18 stimulation; therefore, it is assumed that IL-18 → NIK → NF-κB → BNP further affected the H9c2 hypertrophy. We propose this hypertrophic signaling pathway in H9c2 cardiomyoblasts.

In previous studies, EGCG blocked the activation of NF-κB by inhibiting the activity of IKK [25]. In addition, ventricular hypertrophy induced by transverse abdominal aortic constriction in rats with hypertension could be attenuated by EGCG, which inhibits the activation of mitogen-activated protein kinase (MAPK) [26]. We found that EGCG could reduce IL-18-induced NF-κB activation, which subsequently reduced the expression of BNP, and reduced cardiomyoblast hypertrophy as seen by flow cytometry. Previous studies have highlighted that lipopolysaccharide-induced myocardial hypertrophy was associated with a decrease in the oxidation of fatty acids and an increase in the utilization of glucose [27]. Myocardial hypertrophy can be caused by activation of NF-κB and reduced regulation of fatty acid oxidation. In the present study, we used L-165,041 to activate PPARδ and investigated whether this reduced myocardial hypertrophy. Activation of PPARδ in H9c2 cardiomyoblasts activates expression of PPARα target genes involved in fatty acid utilization in cardiaomyocytes [28]. Therefore, H9c2 cardiomyoblasts were selected for the experiments to obtain results specific to the PPARδ expression. We found that the addition of L-165,041 also inhibited the activation of NF-κB in IL-18-induced H9c2 hypertrophy.

HTR2B are important regulators of heart development and function [8, 9]. Overexpressing the 5-HT Gq-coupled HTR2B induces mitochondrial proliferation and myocardial hypertrophy in rats [10]. Moreover, HTR2B deficiency can cause dilated cardiomyopathy or myofibrillar breakdown [29]. In addition to the effects of HTR2B upregulation in cardiomyocytes, the expression of BNP is regulated through mechanical stress enhanced by HTR2B, which is mainly effected by NF-κB [11]. Therefore, for cardiomyopathy caused by hypertension, reducing the expression of HTR2B may provide a new approach to treatment of myocardial hypertrophy and consequent avoidance of this cause of heart failure. Under the stimulation of IL-18, activation of HTR2B might be directly or indirectly affected to activate the expression of NF-κB, which leads to an upregulation of BNP, and consequently, H9c2 hypertrophy. Therefore, HTR2B might play an important key role in the process of IL-18-induced myocardial hypertrophy. We used flow cytometry to determine whether pretreatment with SB215505 blunted the IL-18-induced hypertrophy of H9c2 and found that SB215505 attenuated the cell hypertrophy significantly. Therefore, we confirmed that HTR2B plays a critical role in the activation of NF-κB and BNP, and thus plays an integral role in IL-18-induced H9c2 cardiomyoblast hypertrophy. In previous studies, IL-18 binding protein (IL-18BP) acted as an antagonist of IL-18 to prevent some of the inflammatory responses induced by IL-18, and reverse damage to the myocardium [30]. In the present study, we found IL-18 upregulated HTR2B and played a critical role in myocardial hypertrophy. We consider myocardial hypertrophy and remodeling are two different concepts, and that inhibiting hypertrophy will lead to a compensatory effect of remodeling.

Here, we found that EGCG and L-165,041 attenuated the upregulation of IL-18-induced HTR2B expression by inhibiting HTR2B signaling. By pretreatment with the HTR2B antagonist SB215505, which inhibits the downstream signaling of HTR2B, the effect of EGCG and L-165,041 on inhibiting the upregulation of HTR2B was not augmented or diminished. This suggests that EGCG, L-165,041, and SB215505 may affect the same signaling pathway. Because SB215505 may not have absolute specificity for HTR2B, particularly at the concentration used (0.1 μM), we also inhibited HTR2B with siRNA to provide a control. We found that in the H9c2 stimulated with IL-18 for 18 h and pretreated with siHTR2B, the expression of MMP-3 and MMP-9 was decreased significantly. Neither EGCG nor L-165,041 augmented this effect, possibly because EGCG and L-165,041 downregulate the expression of MMP-3 and MMP-9 through a HTR2B-dependent pathway.

Conclusions

IL-18 and HTR2B play critical roles in cardiomyoblast hypertrophy. EGCG and L-165,041 inhibit the expression of HTR2B and augment remodeling of H9c2 cardiomyoblasts, possibly mediated by MMP-3 and MMP-9. A new approach to treatment of myocardial hypertrophy and avoidance of consequent heart failure by targeting HTR2B seems plausible.

Figure 1

Pretreatment with 20 mM ECGC and 2 mM L-165,041 attenuated IL-18-induced inflammatory markers in H9c2 cardiomyoblasts. (A) NIK production significantly was inhibited by EGCG or L-165,041 (n = 4). (B) EGCG and L-165,041 significantly attenuated IL-18-induced nuNF-κB upregulation (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18 alone. ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; NIK, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inducing kinase; nuNF-κB, nuclear NF-κB; SEM: standard error of mean.
Pretreatment with 20 mM ECGC and 2 mM L-165,041 attenuated IL-18-induced inflammatory markers in H9c2 cardiomyoblasts. (A) NIK production significantly was inhibited by EGCG or L-165,041 (n = 4). (B) EGCG and L-165,041 significantly attenuated IL-18-induced nuNF-κB upregulation (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18 alone. ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; NIK, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inducing kinase; nuNF-κB, nuclear NF-κB; SEM: standard error of mean.

Figure 2

Effects of 20 mM ECGC and 2 mM L-165,041 treatment on IL-18-induced BNP mRNA and protein expressions in H9c2 cardiomyoblasts. Treatment with IL-18 significantly increased the BNP (n = 4) (A) mRNA and (B) protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. BNP, brain natriuretic peptide; ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SEM: standard error of mean.
Effects of 20 mM ECGC and 2 mM L-165,041 treatment on IL-18-induced BNP mRNA and protein expressions in H9c2 cardiomyoblasts. Treatment with IL-18 significantly increased the BNP (n = 4) (A) mRNA and (B) protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. BNP, brain natriuretic peptide; ECGC (E), epigallocatechin gallate; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SEM: standard error of mean.

Figure 3

Pretreatment with 20 μM EGCG and 2 μM L-165,041 inhibited IL-18-induced HTR2B upregulation. (A) H9c2 cardiomyoblasts treated with IL-18 for up to 24 h, IL-18-induced HTR2B protein expression. Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 4). (B) Serotonin (S) alone (10 μM) significantly increased the expression of HTR2B mRNA. H9c2 were treated IL-18 for 18 h. Pretreatment with EGCG and L-165,041 inhibited IL-18-induced HTR2B mRNA upregulation (n = 5) and (C) HTR2B protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared to IL-18 treatment alone. HTR2B, 5-hydroxytryptamine receptor 2B; ECGC (E), epigallocatechin gallate; IL-18, Interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARδ) agonist; SB215505 (SB), HTR2B antagonist; serotonin, 5-hydroxytryptamine.
Pretreatment with 20 μM EGCG and 2 μM L-165,041 inhibited IL-18-induced HTR2B upregulation. (A) H9c2 cardiomyoblasts treated with IL-18 for up to 24 h, IL-18-induced HTR2B protein expression. Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 4). (B) Serotonin (S) alone (10 μM) significantly increased the expression of HTR2B mRNA. H9c2 were treated IL-18 for 18 h. Pretreatment with EGCG and L-165,041 inhibited IL-18-induced HTR2B mRNA upregulation (n = 5) and (C) HTR2B protein expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 when compared to IL-18 treatment alone. HTR2B, 5-hydroxytryptamine receptor 2B; ECGC (E), epigallocatechin gallate; IL-18, Interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARδ) agonist; SB215505 (SB), HTR2B antagonist; serotonin, 5-hydroxytryptamine.

Figure 4

Pretreatment with 0.1 mM SB215505, 20 mM EGCG, and 2 mM L-165,041 inhibited IL-18-induced H9c2 hypertrophy. H9c2 hypertrophy was inhibited by SB215505, EGCG, and L-165,041, as analyzed by flow cytometry. FSC reflects the relative size of cells. The relative average size for control was 282; IL-18 was 354; IL-18 + SB was 304; IL-18 + SB + EGCG was 272; and IL-18 + SB + L-165,041 was 273 (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. ECGC (E), epigallocatechin gallate; FSC, forward scatter; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SB215505 (SB), HTR2B antagonist; SEM, standard error of mean; SSC, side scatter. HTR2B, 5-hydroxytryptamine receptor 2B.
Pretreatment with 0.1 mM SB215505, 20 mM EGCG, and 2 mM L-165,041 inhibited IL-18-induced H9c2 hypertrophy. H9c2 hypertrophy was inhibited by SB215505, EGCG, and L-165,041, as analyzed by flow cytometry. FSC reflects the relative size of cells. The relative average size for control was 282; IL-18 was 354; IL-18 + SB was 304; IL-18 + SB + EGCG was 272; and IL-18 + SB + L-165,041 was 273 (n = 4). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18. ECGC (E), epigallocatechin gallate; FSC, forward scatter; IL-18, interleukin-18; L-165,041 (L), peroxisome proliferator-activated receptor delta (PPARd) agonist; SB215505 (SB), HTR2B antagonist; SEM, standard error of mean; SSC, side scatter. HTR2B, 5-hydroxytryptamine receptor 2B.

Figure 5

siHTR2B significantly attenuated IL-18-induced HTR2B expression. The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). (A) Pretreatment with siHTR2B significantly decreased HTR2B mRNA expression (n = 5). Differences from control were assessed using a Mann–Whitney U test. (B) Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 5). Pretreatment with siHTR2B significantly inhibited IL-18-induced HTR2B protein expression. Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; SEM, standard error of mean.
siHTR2B significantly attenuated IL-18-induced HTR2B expression. The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines) ± SEM (error bars). (A) Pretreatment with siHTR2B significantly decreased HTR2B mRNA expression (n = 5). Differences from control were assessed using a Mann–Whitney U test. (B) Total protein was extracted and analyzed by western blotting for HTR2B protein expression (n = 5). Pretreatment with siHTR2B significantly inhibited IL-18-induced HTR2B protein expression. Differences between groups were assessed using a Kruskal–Wallis test. **P < 0.01 when compared with control. #P < 0.05 when compared with IL-18. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; SEM, standard error of mean.

Figure 6

Pretreatment with siHTR2B significantly attenuated IL-18-induced MMP-3 and MMP-9 expression. Total protein was extracted and analyzed by western blotting for MMP-3 and MMP-9 expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines; dashed, MMP-3; solid, MMP-9) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18 alone. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; MMP-3, matrix metalloproteinase-3; MMP-9, matrix metalloproteinase-9; SEM, standard error of mean.
Pretreatment with siHTR2B significantly attenuated IL-18-induced MMP-3 and MMP-9 expression. Total protein was extracted and analyzed by western blotting for MMP-3 and MMP-9 expression (n = 5). The data are normalized and shown as individual data (solid circles) and as mean (horizontal lines; dashed, MMP-3; solid, MMP-9) ± SEM (error bars). Differences between groups were assessed using a Kruskal–Wallis test. *P < 0.05 and **P < 0.01 when compared with control. #P < 0.05 and ##P < 0.01 when compared with IL-18 alone. HTR2B, 5-hydroxytryptamine receptor 2B; IL-18, interleukin-18; MMP-3, matrix metalloproteinase-3; MMP-9, matrix metalloproteinase-9; SEM, standard error of mean.

Stroumpoulis KI, Pantazopoulos IN, Xanthos TT. Hypertrophic cardiomyopathy and sudden cardiac death. World J Cardiol. 2010; 2:289–98. StroumpoulisKI PantazopoulosIN XanthosTT Hypertrophic cardiomyopathy and sudden cardiac death World J Cardiol 2010 2 289 98 10.4330/wjc.v2.i9.289299882921160605 Search in Google Scholar

Li J, Umar S, Amjedi M, Iorga A, Sharma S, Nadadur RD, et al. New frontiers in heart hypertrophy during pregnancy. Am J Cardiovasc Dis. 2012; 2:192–207. LiJ UmarS AmjediM IorgaA SharmaS NadadurRD New frontiers in heart hypertrophy during pregnancy Am J Cardiovasc Dis 2012 2 192 207 Search in Google Scholar

O’Brien LC, Mezzaroma E, Van Tassell BW, Marchetti C, Carbone S, Abbate A, Toldo S. Interleukin-18 as a therapeutic target in acute myocardial infarction and heart failure. Mol Med. 2014; 20:221–9. O’BrienLC MezzaromaE Van TassellBW MarchettiC CarboneS AbbateA ToldoS Interleukin-18 as a therapeutic target in acute myocardial infarction and heart failure Mol Med 2014 20 221 9 10.2119/molmed.2014.00034406926924804827 Search in Google Scholar

Wang M, Markel TA, Meldrum DR. Interleukin 18 in the heart. Shock. 2008; 30:3–10. WangM MarkelTA MeldrumDR Interleukin 18 in the heart Shock 2008 30 3 10 10.1097/SHK.0b013e318160f21518562922 Search in Google Scholar

Hartford M, Wiklund O, Hultén LM, Persson A, Karlsson T, Herlitz J, et al. Interleukin-18 as a predictor of future events in patients with acute coronary syndromes. Arterioscler Thromb Vasc Biol. 2010; 30:2039–46. HartfordM WiklundO HulténLM PerssonA KarlssonT HerlitzJ Interleukin-18 as a predictor of future events in patients with acute coronary syndromes Arterioscler Thromb Vasc Biol 2010 30 2039 46 10.1161/ATVBAHA.109.20269720689079 Search in Google Scholar

Badimon L. Interleukin-18: a potent pro-inflammatory cytokine in atherosclerosis. Cardiovasc Res. 2012; 96:172–5. BadimonL Interleukin-18: a potent pro-inflammatory cytokine in atherosclerosis Cardiovasc Res 2012 96 172 5 10.1093/cvr/cvs22622922166 Search in Google Scholar

Colston JT, Boylston WH, Feldman MD, Jenkinson CP, de la Rosa SD, Barton A, et al. Interleukin-18 knockout mice display maladaptive cardiac hypertrophy in response to pressure overload. Biochem Biophys Res Commun. 2007; 354:552–8. ColstonJT BoylstonWH FeldmanMD JenkinsonCP de la RosaSD BartonA Interleukin-18 knockout mice display maladaptive cardiac hypertrophy in response to pressure overload Biochem Biophys Res Commun 2007 354 552 8 10.1016/j.bbrc.2007.01.030184763617250807 Search in Google Scholar

Nebigil CG, Choi D-S, Dierich A, Hickel P, Le Meur M, Messaddeq N, et al. Serotonin 2B receptor is required for heart development. Proc Natl Acad Sci U S A. 2000; 97:9508–13. NebigilCG ChoiD-S DierichA HickelP Le MeurM MessaddeqN Serotonin 2B receptor is required for heart development Proc Natl Acad Sci U S A 2000 97 9508 13 10.1073/pnas.97.17.95081689510944220 Search in Google Scholar

Janssen W, Schymura Y, Novoyatleva T, Kojonazarov B, Boehm M, Wietelmann A, et al. 5-HT2B receptor antagonists inhibit fibrosis and protect from RV heart failure. Biomed Res Int. 2015; 2015:438403. doi: 10.1155/2015/438403 JanssenW SchymuraY NovoyatlevaT KojonazarovB BoehmM WietelmannA 5-HT2B receptor antagonists inhibit fibrosis and protect from RV heart failure Biomed Res Int 2015 2015 438403 10.1155/2015/438403 431257425667920 Ouvrir le DOISearch in Google Scholar

Nebigil CG, Jaffré F, Messaddeq N, Hickel P, Monassier L, Launay JM, Maroteaux L. Overexpression of the serotonin 5-HT2B receptor in heart leads to abnormal mitochondrial function and cardiac hypertrophy. Circulation. 2003; 107:3223–9. NebigilCG JaffréF MessaddeqN HickelP MonassierL LaunayJM MaroteauxL Overexpression of the serotonin 5-HT2B receptor in heart leads to abnormal mitochondrial function and cardiac hypertrophy Circulation 2003 107 3223 9 10.1161/01.CIR.0000074224.57016.0112810613 Search in Google Scholar

Liang Y-J, Lai L-P, Wang B-W, Juang S-J, Chang C-M, Leu J-G, Shyu K-G. Mechanical stress enhances serotonin 2B receptor modulating brain natriuretic peptide through nuclear factor-κB in cardiomyocytes. Cardiovasc Res. 2006; 72:303–12. LiangY-J LaiL-P WangB-W JuangS-J ChangC-M LeuJ-G ShyuK-G Mechanical stress enhances serotonin 2B receptor modulating brain natriuretic peptide through nuclear factor-κB in cardiomyocytes Cardiovasc Res 2006 72 303 12 10.1016/j.cardiores.2006.08.00316962085 Search in Google Scholar

Jaffré F, Bonnin P, Callebert J, Debbabi H, Setola V, Doly S, et al. Serotonin and angiotensin receptors in cardiac fibroblasts coregulate adrenergic-dependent cardiac hypertrophy. Circ Res. 2009; 104:113–23. JaffréF BonninP CallebertJ DebbabiH SetolaV DolyS Serotonin and angiotensin receptors in cardiac fibroblasts coregulate adrenergic-dependent cardiac hypertrophy Circ Res 2009 104 113 23 10.1161/CIRCRESAHA.108.180976 Search in Google Scholar

Hamblin M, Chang L, Fan Y, Zhang J, Chen YE. PPARs and the cardiovascular system. Antioxid Redox Signal. 2009; 11:1415–52. HamblinM ChangL FanY ZhangJ ChenYE PPARs and the cardiovascular system Antioxid Redox Signal 2009 11 1415 52 10.1089/ars.2008.2280 Search in Google Scholar

Zheng Y, Song HJ, Kim CH, Kim HS, Kim E-G, Sachinidis A, Ahn HY. Inhibitory effect of epigallocatechin 3-O-gallate on vascular smooth muscle cell hypertrophy induced by angiotensin II. J Cardiovasc Pharmacol. 2004; 43:200–8. ZhengY SongHJ KimCH KimHS KimE-G SachinidisA AhnHY Inhibitory effect of epigallocatechin 3-O-gallate on vascular smooth muscle cell hypertrophy induced by angiotensin II J Cardiovasc Pharmacol 2004 43 200 8 10.1097/00005344-200402000-00006 Search in Google Scholar

Won S-M, Park Y-H, Kim H-J, Park K-M, Lee W-J. Catechins inhibit angiotensin II-induced vascular smooth muscle cell proliferation via mitogen-activated protein kinase pathway. Exp Mol Med. 2006; 38:525–34. WonS-M ParkY-H KimH-J ParkK-M LeeW-J Catechins inhibit angiotensin II-induced vascular smooth muscle cell proliferation via mitogen-activated protein kinase pathway Exp Mol Med 2006 38 525 34 10.1038/emm.2006.62 Search in Google Scholar

Villeneuve C, Caudrillier A, Ordener C, Pizzinat N, Parini A, Mialet-Perez J. Dose-dependent activation of distinct hypertrophic pathways by serotonin in cardiac cells. Am J Physiol Heart Circ Physiol. 2009; 297:H821–8. VilleneuveC CaudrillierA OrdenerC PizzinatN PariniA Mialet-PerezJ Dose-dependent activation of distinct hypertrophic pathways by serotonin in cardiac cells Am J Physiol Heart Circ Physiol 2009 297 H821 8 10.1152/ajpheart.00345.2009 Search in Google Scholar

Kimes BW, Brandt BL. Properties of a clonal muscle cell line from rat heart. Exp Cell Res. 1976; 98:367–81. KimesBW BrandtBL Properties of a clonal muscle cell line from rat heart Exp Cell Res 1976 98 367 81 10.1016/0014-4827(76)90447-X Search in Google Scholar

Ayme-Dietrich E, Aubertin-Kirch G, Maroteaux L, Monassier L. Cardiovascular remodeling and the peripheral serotonergic system. Arch Cardiovasc Dis. 2017; 110:51–9. Ayme-DietrichE Aubertin-KirchG MaroteauxL MonassierL Cardiovascular remodeling and the peripheral serotonergic system Arch Cardiovasc Dis 2017 110 51 9 10.1016/j.acvd.2016.08.00228017279 Search in Google Scholar

Cheng K-C, Chang W-T, Li Y, Cheng Y-Z, Cheng J-T, Chen Z-C. GW0742 activates peroxisome proliferator-activated receptor δ to reduce free radicals and alleviate cardiac hypertrophy induced by hyperglycemia in cultured H9c2 cells. J Cell Biochem. 2018; 119:9532–42. ChengK-C ChangW-T LiY ChengY-Z ChengJ-T ChenZ-C GW0742 activates peroxisome proliferator-activated receptor δ to reduce free radicals and alleviate cardiac hypertrophy induced by hyperglycemia in cultured H9c2 cells J Cell Biochem 2018 119 9532 42 10.1002/jcb.2727030129179 Search in Google Scholar

Chandrasekar B, Mummidi S, Claycomb WC, Mestril R, Nemer M. Interleukin-18 is a pro-hypertrophic cytokine that acts through a phosphatidylinositol 3-kinase-phosphoinositide-dependent kinase-1-Akt-GATA4 signaling pathway in cardiomyocytes. J Biol Chem. 2005; 280:4553–67. ChandrasekarB MummidiS ClaycombWC MestrilR NemerM Interleukin-18 is a pro-hypertrophic cytokine that acts through a phosphatidylinositol 3-kinase-phosphoinositide-dependent kinase-1-Akt-GATA4 signaling pathway in cardiomyocytes J Biol Chem 2005 280 4553 67 10.1074/jbc.M41178720015574430 Search in Google Scholar

Chandrasekar B, Boylston WH, Venkatachalam K, Webster NJ, Prabhu SD, Valente AJ. Adiponectin blocks interleukin-18-mediated endothelial cell death via APPL1-dependent AMP-activated protein kinase (AMPK) activation and IKK/NF-κB/PTEN suppression. J Biol Chem. 2008; 283:24889–98. ChandrasekarB BoylstonWH VenkatachalamK WebsterNJ PrabhuSD ValenteAJ Adiponectin blocks interleukin-18-mediated endothelial cell death via APPL1-dependent AMP-activated protein kinase (AMPK) activation and IKK/NF-κB/PTEN suppression J Biol Chem 2008 283 24889 98 10.1074/jbc.M804236200325983118632660 Search in Google Scholar

Freund C, Schmidt-Ullrich R, Baurand A, Dunger S, Schneider W, Loser P, et al. Requirement of nuclear factor-κB in angiotensin II– and isoproterenol-induced cardiac hypertrophy in vivo. Circulation. 2005; 111:2319–25. FreundC Schmidt-UllrichR BaurandA DungerS SchneiderW LoserP Requirement of nuclear factor-κB in angiotensin II– and isoproterenol-induced cardiac hypertrophy in vivo Circulation 2005 111 2319 25 10.1161/01.CIR.0000164237.58200.5A15870116 Search in Google Scholar

Nakagawa Y, Nishikimi T, Kuwahara K. Atrial and brain natriuretic peptides: hormones secreted from the heart. Peptides. 2019; 111:18–25. doi: 10.1016/j.peptides.2018.05.012 NakagawaY NishikimiT KuwaharaK Atrial and brain natriuretic peptides: hormones secreted from the heart Peptides 2019 111 18 25 10.1016/j.peptides.2018.05.012 29859763 Ouvrir le DOISearch in Google Scholar

Murray DR, Mummidi S, Valente AJ, Yoshida T, Somanna NK, Delafontaine P, et al. β2 adrenergic activation induces the expression of IL-18 binding protein, a potent inhibitor of isoproterenol induced cardiomyocyte hypertrophy in vitro and myocardial hypertrophy in vivo. J Mol Cell Cardiol. 2012; 52:206–18. MurrayDR MummidiS ValenteAJ YoshidaT SomannaNK DelafontaineP β2 adrenergic activation induces the expression of IL-18 binding protein, a potent inhibitor of isoproterenol induced cardiomyocyte hypertrophy in vitro and myocardial hypertrophy in vivo J Mol Cell Cardiol 2012 52 206 18 10.1016/j.yjmcc.2011.09.022324602622004899 Search in Google Scholar

Lagha AB, Grenier D. Tea polyphenols inhibit the activation of NF-κB and the secretion of cytokines and matrix metalloproteinases by macrophages stimulated with Fusobacterium nucleatum. Sci Rep. 2016; 6:34520. doi: 10.1038/srep34520 LaghaAB GrenierD Tea polyphenols inhibit the activation of NF-κB and the secretion of cytokines and matrix metalloproteinases by macrophages stimulated with Fusobacterium nucleatum Sci Rep 2016 6 34520 10.1038/srep34520 504613427694921 Ouvrir le DOISearch in Google Scholar

Hao J, Kim C-H, Ha T-S, Ahn H-Y. Epigallocatechin-3 gallate prevents cardiac hypertrophy induced by pressure overload in rats. J Vet Sci. 2007; 8:121–9. HaoJ KimC-H HaT-S AhnH-Y Epigallocatechin-3 gallate prevents cardiac hypertrophy induced by pressure overload in rats J Vet Sci 2007 8 121 9 10.4142/jvs.2007.8.2.121287270917519564 Search in Google Scholar

Planavila A, Laguna JC, Vázquez-Carrera M. Nuclear factor-κB activation leads to down-regulation of fatty acid oxidation during cardiac hypertrophy. J Biol Chem. 2005; 280:17464–71. PlanavilaA LagunaJC Vázquez-CarreraM Nuclear factor-κB activation leads to down-regulation of fatty acid oxidation during cardiac hypertrophy J Biol Chem 2005 280 17464 71 10.1074/jbc.M41422020015728586 Search in Google Scholar

Planavila A, Rodríguez-Calvo R, Jové M, Michalik L, Wahli W, Laguna JC, Vázquez-Carrera M. Peroxisome proliferator-activated receptor β/δ activation inhibits hypertrophy in neonatal rat cardiomyocytes. Cardiovasc Res. 2005; 65:832–41. PlanavilaA Rodríguez-CalvoR JovéM MichalikL WahliW LagunaJC Vázquez-CarreraM Peroxisome proliferator-activated receptor β/δ activation inhibits hypertrophy in neonatal rat cardiomyocytes Cardiovasc Res 2005 65 832 41 10.1016/j.cardiores.2004.11.01115721863 Search in Google Scholar

Nebigil CG, Maroteaux L. Functional consequence of serotonin/5-HT2B receptor signaling in heart: role of mitochondria in transition between hypertrophy and heart failure? Circulation. 2003; 108:902–8. NebigilCG MaroteauxL Functional consequence of serotonin/5-HT2B receptor signaling in heart: role of mitochondria in transition between hypertrophy and heart failure? Circulation 2003 108 902 8 10.1161/01.CIR.0000081520.25714.D912925446 Search in Google Scholar

Dinarello CA, Novick D, Kim S, Kaplanski G. Interleukin-18 and IL-18 binding protein. Front Immunol. 2013; 4:289. doi: 10.3389/fimmu.2013.00289 DinarelloCA NovickD KimS KaplanskiG Interleukin-18 and IL-18 binding protein Front Immunol 2013 4 289 10.3389/fimmu.2013.00289 379255424115947 Ouvrir le DOISearch in Google Scholar

Articles recommandés par Trend MD

Planifiez votre conférence à distance avec Sciendo