β-arrestin2 Mediates the Arginine Vasopressin-Induced Expression of IL-1β in Murine Hearts

Background: Circulating levels of arginine vasopressin (AVP) are elevated during cardiac stress and this could be a factor in cardiac inflammation and fibrosis. Herein, we studied the effects of AVP on interleukin-1 $\beta{}$ (IL-1 $\beta{}$ ) production and the role(s) of $\beta{}$ -arrestin2-dependent signaling in murine heart. Methods: The levels of IL-1 $\beta{}$ mRNA and protein in adult rat cardiofibroblasts (ARCFs) was measured using quantitative PCR and ELISA, respectively. The activity of $\beta{}$ -arrestin2 was manipulated using either pharmacologic inhibitors or through recombinant $\beta{}$ -arrestin2 over-expression. These experiments were conducted to determine the roles of $\beta{}$ -arrestin2 in the regulation of AVP-induced IL-1 $\beta{}$ and NLRP3 inflammasome production. The phosphorylation and activation of NF- $\kappa{}$ B induced by AVP was measured by immunoblotting. $\beta{}$ -arrestin2 knockout (KO) mice were used to investigate whether $\beta{}$ -arrestin2 mediated the AVP-induced production of IL-1 $\beta{}$ and NLRP3, as well as the phosphorylation of the NF- $\kappa{}$ B p65 subunitin mouse myocardium. Prism GraphPad software(version 8.0), was used for all statistical analyses. Results: AVP induced the expression of IL-1 $\beta{}$ in a time-dependent manner in ARCFs but not in cultured adult rat cardiomyocytes (ARCMs). The inhibition of NF- $\kappa{}$ B with pyrrolidinedithiocarbamic acid (PDTC) prevented the AVP-induced phosphorylation of NF- $\kappa{}$ B and production of IL-1 $\beta{}$ and NLRP3 in ARCFs. The deletion of $\beta{}$ -arrestin2 blocked the phosphorylation of p65 and the expression of NLRP3 and IL-1 $\beta{}$ induced by AVP in both mouse hearts and in ARCFs. Conclusions: AVP promotes IL-1 $\beta{}$ expression through $\beta{}$ -arrestin2-mediated NF- $\kappa{}$ B signaling in murine heart.

Keywords

arginine vasopressin

β-arrestin 2

NF-κB p65

NLRP3 inflammasome

interlukin-1β

inflammation

murine

Funding

81770400/National Natural Science Foundation of China

JCZ18131/Nantong Municipal Science and Technology

JC2019133/Nantong Municipal Science and Technology

Figures

Fig. 1.

Previous article in this issue

Next article in this issue

Fig. 1.

AVP induced expression of the NLRP3 inflammasome and IL-1 $\beta{}$ in mouse myocardium following AVP administration. (A,B) Representative images of wild type mice injected with 2 $\mu{}$ g/g body weight of AVP or 0.9% of NaCl via the tail vein. Frozen sections were prepared from dissected heart tissue to determine the protein level of the NLRP3 inflammasome and IL-1 $\beta{}$ 6 h after dosing with NaCl or AVP. (C,D) Fluorescent optical density was determined and data expressed as mean $\pm{}$ S.E.M. of 3 separate animals with triplicate tissue sections, **p $<$ 0.01, ***p $<$ 0.001. one-way ANOVA test, followed by Bonferroni’s post hoc test. Note: Non-specific binding staining was examined using mouse IgG. Scale bar = 50 $\mu{}$ m.

1 of 7

Front. Biosci. (Landmark Ed) Print ISSN 2768-6701 Electronic ISSN 2768-6698