title: Gelatinases promote calcification of vascular smooth muscle cells by up-regulating bone morphogenetic protein-2.
abstract: Matrix metalloproteinase-2 (MMP-2), also known as gelatinase A, is involved in vascular calcification. Another member of gelatinases is MMP-9 (gelatinase B). However, the role of gelatinases in the pathogenesis of vascular calcification is not well understood. The current study aims to clarify the relationship between gelatinases and vascular calcification and to elucidate the underlying mechanism. Beta-glycerophosphate (β-GP) was used to induce calcification of vascular smooth muscle cells (VSMCs) with or without 2-[[(4-Phenoxyphenyl)sulfonyl]methyl]-thiirane (SB-3CT), a specific gelatinases inhibitor. Levels of calcification were determined by assessing calcium content and calcification area of VSMCs. Phenotype transition of VSMCs was observed by assessing expressions of alkaline phosphatase (ALP), smooth muscle α-actin (SM-α-actin) and desmin. Gelatin zymography was applied to determine the activities of gelatinases, and western blot was applied to determine expressions of gelatinases, bone morphogenetic protein-2 (BMP-2), Runt-related transcription factor 2 (RUNX2) and msh homeobox homolog 2 (Msx-2). Gelatinases inhibition by SB-3CT alleviated calcification and phenotype transition of VSMCs induced by β-GP. Increased gelatinases expression and active MMP-2 were observed in calcifying VSMCs. Gelatinases inhibition reduced expression of RUNX2, Msx-2 and BMP-2. BMP-2 treatment increased expressions of RUNX2 and Msx-2, while noggin, an antagonist of BMP-2, decreased expressions of RUNX2 and Msx-2. Gelatinases promote vascular calcification by upregulating BMP-2 which induces expression of RUNX2 and Msx-2, two proteins associated with phenotype transition of VSMCs in vascular calcification. Interventions targeting gelatinases inhibition might be a proper candidate for ameliorating vascular calcification.
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: zyg307@126.com.
authors.forename: Yong-Gang
authors.initials: YG
authors.lastname: Zhao
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: mfx2143@163.com.
authors.forename: Fan-Xing
authors.initials: FX
authors.lastname: Meng
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: drli715@163.com.
authors.forename: Bing-Wei
authors.initials: BW
authors.lastname: Li
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: shengbiomed@163.com.
authors.forename: You-Ming
authors.initials: YM
authors.lastname: Sheng
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: mingmingliu@imc.pumc.edu.cn.
authors.forename: Ming-Ming
authors.initials: MM
authors.lastname: Liu
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: bw20032007@126.com.
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: lihw66@imc.pumc.edu.cn.
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Li
authors.affiliation: Institute of Microcirculation and Key Laboratory, Ministry of Health, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China. Electronic address: xiurj@imc.pumc.edu.cn.
authors.forename: Rui-Juan
authors.initials: RJ
authors.lastname: Xiu
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism*; D002478:Cells, Cultured; D018093:Gelatinases / Q000378:metabolism*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000378:metabolism* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015854:Up-Regulation; D061205:Vascular Calcification / Q000378:metabolism* / Q000473:pathology*
chemical_list: C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D018093:Gelatinases
keywords: Bone morphogenetic protein-2; Matrix metalloproteinase-2; Runt-related transcription factor 2; Vascular calcification; Vascular smooth muscle cell