Xiuli Zhang1,2,3, Yining Li1,2,3, Pingzhen Yang1,2,3, Xiaoyu Liu1,2,3, Lihe Lu4, Yanting Chen4, Xinglong Zhong1,2,3, Zehua Li1,2,3, Hailin Liu1,2,3, Caiwen Ou1,2,3, Jianyun Yan1,2,3, Minsheng Chen1,2,3. 1. From the Department of Cardiology, Laboratory of Heart Center, Zhujiang Hospital, Southern Medical University, China (X.Z., Y.L., P.Y., X.L., X.Z., Z.L., H.L., C.O., J.Y., M.C.). 2. Guangdong Provincial Biomedical Engineering Technology Research Center for Cardiovascular Disease, China (X.Z., Y.L., P.Y., X.L., X.Z., Z.L., H.L., C.O., J.Y., M.C.). 3. Sino-Japanese Cooperation Platform for Translational Research in Heart Failure, China (X.Z., Y.L., P.Y., X.L., X.Z., Z.L., H.L., C.O., J.Y., M.C.). 4. Department of Pathophysiolgy, Zhongshan Medical School, Sun Yat-Sen University, China (L.L., Y.C.).
Abstract
OBJECTIVES: Vascular calcification is highly prevalent in patients with chronic kidney disease. Increased plasma trimethylamine N-oxide (TMAO), a gut microbiota-dependent product, concentrations are found in patients undergoing hemodialysis. However, a clear mechanistic link between TMAO and vascular calcification is not yet established. In this study, we investigate whether TMAO participates in the progression of vascular calcification using in vitro, ex vivo, and in vivo models. Approach and Results: Alizarin red staining revealed that TMAO promoted calcium/phosphate-induced calcification of rat and human vascular smooth muscle cells in a dose-dependent manner, and this was confirmed by calcium content assay. Similarly, TMAO upregulated the expression of bone-related molecules including Runx2 (Runt-related transcription factor 2) and BMP2 (bone morphogenetic protein-2), suggesting that TMAO promoted osteogenic differentiation of vascular smooth muscle cells. In addition, ex vivo study also showed the positive regulatory effect of TMAO on vascular calcification. Furthermore, we found that TMAO accelerated vascular calcification in rats with chronic kidney disease, as indicated by Mico-computed tomography analysis, alizarin red staining and calcium content assay. By contrast, reducing TMAO levels by antibiotics attenuated vascular calcification in chronic kidney disease rats. Interestingly, TMAO activated NLRP3 (nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3) inflammasome and NF-κB (nuclear factor κB) signals during vascular calcification. Inhibition of NLRP3 inflammasome and NF-κB signals attenuated TMAO-induced vascular smooth muscle cell calcification. CONCLUSIONS: This study for the first time demonstrates that TMAO promotes vascular calcification through activation of NLRP3 inflammasome and NF-κB signals, suggesting the potential link between gut microbial metabolism and vascular calcification. Reducing the levels of TMAO could become a potential treatment strategy for vascular calcification in chronic kidney disease.
OBJECTIVES:Vascular calcification is highly prevalent in patients with chronic kidney disease. Increased plasma trimethylamine N-oxide (TMAO), a gut microbiota-dependent product, concentrations are found in patients undergoing hemodialysis. However, a clear mechanistic link between TMAO and vascular calcification is not yet established. In this study, we investigate whether TMAO participates in the progression of vascular calcification using in vitro, ex vivo, and in vivo models. Approach and Results:Alizarin red staining revealed that TMAO promoted calcium/phosphate-induced calcification of rat and human vascular smooth muscle cells in a dose-dependent manner, and this was confirmed by calcium content assay. Similarly, TMAO upregulated the expression of bone-related molecules including Runx2 (Runt-related transcription factor 2) and BMP2 (bone morphogenetic protein-2), suggesting that TMAO promoted osteogenic differentiation of vascular smooth muscle cells. In addition, ex vivo study also showed the positive regulatory effect of TMAO on vascular calcification. Furthermore, we found that TMAO accelerated vascular calcification in rats with chronic kidney disease, as indicated by Mico-computed tomography analysis, alizarin red staining and calcium content assay. By contrast, reducing TMAO levels by antibiotics attenuated vascular calcification in chronic kidney diseaserats. Interestingly, TMAO activated NLRP3 (nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3) inflammasome and NF-κB (nuclear factor κB) signals during vascular calcification. Inhibition of NLRP3 inflammasome and NF-κB signals attenuated TMAO-induced vascular smooth muscle cell calcification. CONCLUSIONS: This study for the first time demonstrates that TMAO promotes vascular calcification through activation of NLRP3 inflammasome and NF-κB signals, suggesting the potential link between gut microbial metabolism and vascular calcification. Reducing the levels of TMAO could become a potential treatment strategy for vascular calcification in chronic kidney disease.
Authors: Meng Wang; Zeneng Wang; Stanley L Hazen; Dariush Mozaffarian; Yujin Lee; Heidi T M Lai; Marcia C de Oliveira Otto; Rozenn N Lemaitre; Amanda Fretts; Nona Sotoodehnia; Matthew Budoff; Joseph A DiDonato; Barbara McKnight; W H Wilson Tang; Bruce M Psaty; David S Siscovick Journal: Arterioscler Thromb Vasc Biol Date: 2022-08-01 Impact factor: 10.514