Diabetes, Metabolic Syndrome and Obesity (Nov 2020)
Role of Sortilin and Matrix Vesicles in Nϵ-Carboxymethyl-Lysine-Induced Diabetic Atherosclerotic Calcification
Abstract
Lele Jing,1 Lihua Li,2 Xiaomei Ren,3 Zhen Sun,1 Zhengyang Bao,4 Guoyue Yuan,5 Honghua Cai,6 Lin Wang,1 Chen Shao,1 Zhongqun Wang1 1Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, People’s Republic of China; 2Department of Pathology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, People’s Republic of China; 3Department of Geriatrics, Zhongda Hospital Affiliated of Southeast University, Nanjing, People’s Republic of China; 4Department of Internal Medicine, Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University, Wuxi 214000, People’s Republic of China; 5Department of Endocrinology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, People’s Republic of China; 6Department of Burns and Plastic Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, People’s Republic of ChinaCorrespondence: Zhongqun WangDepartment of Cardiology, Affiliated Hospital of Jiangsu University, 438 Jiefang Road, Zhenjiang 212001, People’s Republic of ChinaTel +86 511 85030586Email [email protected] and Aims: To investigate the role of Sortilin and matrix vesicles (MVs) in Nϵ-Carboxymethyl-lysine (CML)-induced diabetic atherosclerotic calcification (AC).Methods: At human level, the correlation between Sortilin and CD9 (marker proteins of MVs) in serum MVs and CML in serum was explored by enzyme-linked immunosorbent assay (ELISA) detection and Pearson correlation analysis. After a diabetic apoE-/- mouse model was constructed, the calcification of aorta and the expressions of related proteins under CML and MVs injection were observed by calcification staining, immunofluorescence staining, and Western blot. MVs levels released by smooth muscle cells (SMCs) under different treatments was detected by nanometer tracking analysis (NTA). After treating SMCs with MVs and Anti-Sortilin, cell calcification was observed by Alizarin red staining.Results: Serological analysis of patients showed that the concentrations of Sortilin and CD9 in serum MVs were positively correlated with the concentration of CML in serum. Animal experiments showed that CML could promote the progression of diabetic AC and the high expression of Sortilin in plaques. Diabetic apoE-/- mouse tail vein injection of CML-induced SMCs-derived MVs obviously aggravated AC. Cell experiment results showed that a high concentration of CML significantly promoted the release of MVs from SMCs. MVs from this source could markedly worsen cell calcification, while the administration of GW4869 (a widely used extracellular vesicles biogenesis inhibitor) significantly reduced cell calcification. Finally, treatment of high concentrations of CML could also promote the recruitment of Sortilin to MVs, and administration of Anti-Sortilin could markedly reduce cell calcification caused by MVs.Conclusion: We proved that CML not only affects the release of MVs from SMCs but also affects the recruitment of Sortilin to MVs, thereby promoting diabetic AC. This discovery may provide a new strategy for targeted prevention of vascular calcification in diabetes.Keywords: Nϵ-Carboxymethyl-lysine, sortilin, matrix vesicles, plaque calcification, diabetes
