Irisin protects against vascular calcification by activating autophagy and inhibiting NLRP3-mediated vascular smooth muscle cell pyroptosis in chronic kidney disease

Abstract

Irisin protects the cardiovascular system against vascular diseases. However, its role in chronic kidney disease (CKD) -associated vascular calcification (VC) and the underlying mechanisms remain unclear. In the present study, we investigated the potential link among Irisin, pyroptosis, and VC under CKD conditions. During mouse vascular smooth muscle cell (VSMC) calcification induced by β-glycerophosphate (β-GP), the pyroptosis level was increased, as evidenced by the upregulated expression of pyroptosis-related proteins (cleaved CASP1, GSDMD-N, and IL1B) and pyroptotic cell death (increased numbers of PI-positive cells and LDH release). Reducing the pyroptosis levels by a CASP1 inhibitor remarkably decreased calcium deposition in β-GP-treated VSMCs. Further experiments revealed that the pyroptosis pathway was activated by excessive reactive oxygen species (ROS) production and subsequent NLR family pyrin domain containing 3 (NLRP3) inflammasome activation in calcified VSMCs. Importantly, Irisin effectively inhibited β-GP-induced calcium deposition in VSMCs in vitro and in mice aortic rings ex vivo. Overexpression of Nlrp3 attenuated the suppressive effect of Irisin on VSMC calcification. In addition, Irisin could induce autophagy and restore autophagic flux in calcified VSMCs. Adding the autophagy inhibitor 3-methyladenine or chloroquine attenuated the inhibitory effect of Irisin on β-GP-induced ROS production, NLRP3 inflammasome activation, pyroptosis, and calcification in VSMCs. Finally, our in vivo study showed that Irisin treatment promoted autophagy, downregulated ROS level and thereby suppressed pyroptosis and medial calcification in aortic tissues of adenine-induced CKD mice. Together, our findings for the first time demonstrated that Irisin protected against VC via inducing autophagy and inhibiting VSMC pyroptosis in CKD, and Irisin might serve as an effective therapeutic agent for CKD-associated VC.

The crosstalk among autophagy, apoptosis, and pyroptosis in cardiovascular disease

“…In vivo studies have shown that Irisin treatment promotes autophagy, downregulates ROS level, and thereby inhibits pyroptosis and medial calcification. This finding suggests that Irisin can protect against VC by inducing autophagy and inhibiting pyroptosis ( 107 , 108 ).…”

Vascular calcification: Molecular mechanisms and therapeutic interventions

“… 139 NLRP3‐mediated VSMC pyroptosis was reported to take part in the molecular protective mechanism of irisin against VC. 140 HASMCs calcification was reported to restain in Atg7−/− animal model after AGEs induction. Atg7 was recognized as an autophagy inhibitor, indicating that autophagy was an important mechanism in diabetes‐related VC.…”

Neuroplasticity to autophagy cross-talk in a therapeutic effect of physical exercises and irisin in ADHD

“…Pang et al showed that exogenous irisin (100 ng/ml, 72 h) leads to an increase in the levels of LC3II and p62, as well as to an increase to LC3II/ LC3I ratio in vascular smooth muscle cells in mice. Notably, this effect goes along to the increase in levels of mRNAs of Map1lc3b, Becn1, Atg7, Atg5, and Lamp1, further highlighting epigenomic effects of irisin (Pang et al, 2022). Pan et al showed that exogenous irisin (20 nM, 48 h) was diminishing the toxic effect of doxorubicin in the culture of endothelial cells.…”