Annexin-Mediated Matrix Vesicle Calcification in Vascular Smooth Muscle Cells
Annexin-Mediated Matrix Vesicle Calcification in Vascular Smooth Muscle Cells
复制标题
DOI：
10.1359/jbmr.080604
复制
发表时间：
2008-11-01
影响因子：
6.2
通讯作者：
Moe, Sharon M.
中科院分区：
文献类型：
--
作者：
Chen, Neal X.;O'Neill, Kalisha D.;Moe, Sharon M.
在骨中，成骨细胞和软骨细胞合成与胶原相互作用以启动钙化的基质囊泡（MV）。已经在人类钙化动脉中识别出MV，但表征不佳。本研究的目的是确定在牛血管平滑肌细胞（BVSMCs）钙化过程中膜联蛋白和Feutin-A在MV形成和活性中的作用。用对照或钙化（高磷）培养基处理BVSMC，通过胶原酶消化分离细胞MV，并通过超离心从培养基中分离分泌的MV。结果表明，碱性磷酸酶（ALP）活性显着增加的MV钙化的BVSMCs相比，非钙化的BVSMCs，是膜联蛋白II和VT含量和Ca-45摄取。我们还确定，钙化的BVSMCs的MV可以矿化I型胶原，但不是II型胶原的细胞在剂量和时间依赖性的方式。通过K201阻断膜联蛋白钙通道活性显著降低ALP活性，并降低MV随后钙化油胶原的能力。无论K201是在MV形成期间还是之后加入。此外。与分泌型MV相比，细胞型MV具有显著增加的钙化油胶原的能力，这可能是因为它们增加的ALP活性和膜联蛋白II含量，但胎球蛋白-A含量低。总之，我们的研究结果表明，在VSMCs的矿化需要两个积极的MV和MV与I型胶原蛋白的相互作用，这两个步骤需要膜联蛋白的活性。
In bone, osteoblasts and chondrocytes synthesize matrix vesicles (MVs) that interact with collagen to initiate calcification. MVs have been identified in human calcified arteries but are poorly Characterized. The objective of this Study is to determine the role of annexins and feutin-A in MV formation and activity during calcification in bovine vascular smooth Muscle cells (BVSMCs). BVSMCs were treated with control or calcification (high phosphorus) media, and cellular MVs were isolated by collagenase digestion and secreted MVs were isolated from cultured media by ultracentrifugation. The results Showed that alkaline phosphatase (ALP) activity was significantly increased in MVs from calcified BVSMCs compared with non-calcified BVSMCs, as was annexin II and VT content and Ca-45 uptake. We also determined that MVs from calcifying BVSMCs could mineralize type I collagen but not type II collagen in the absence of cells in a dose-and time-dependent manner. Blockade of annexin calcium channel activity by K201 significantly decreased ALP activity and reduced the ability of the MVs to subsequently calcify oil collagen. whether the K201 was added during or after MV formation. Furthermore. cellular MVs had significantly increased ability to calcify oil collagen compared with secreted MVs, likely because of their increased ALP activity and annexin II content but low fetuin-A content. In conclusion, our results suggest that mineralization in VSMCs requires both active MVs and an interaction of the MVs with type I collagen, and both steps require annexin activity.