Elastogenic effects of exogenous hyaluronan oligosaccharides on vascular smooth muscle cells
Elastogenic effects of exogenous hyaluronan oligosaccharides on vascular smooth muscle cells
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DOI：
10.1016/j.biomaterials.2006.07.020
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发表时间：
2006-11-01
期刊：
影响因子：
14
通讯作者：
Ramamurthi, Anand
中科院分区：
文献类型：
--
作者：
Joddar, Binata;Ramamurthi, Anand
先前的研究表明，透明质酸（HA），一种糖胺聚糖，可能上调成人血管平滑肌细胞（SMCs）天生缺乏弹性蛋白基质的合成。因此，透明质酸支架可用于再生受损的血管弹性蛋白。在早期的研究中，我们确定非低聚HA的弹性效应是片段大小和/或剂量特异性的。我们目前正在研究外源性HA寡聚物对大鼠主动脉平滑肌细胞（RASMCs）的促弹性作用。RASMCs用纯HA低聚物（4-mers）和酶切长链HA获得的混合物（4-8 -mers）（分子量接近2000 kDa）培养。聚丙烯酰胺凝胶电泳(PAGE)/基质辅助激光解吸/电离光谱飞行时间分析（MALDI-TOF）显示，HA消化液中含有4- 8mers的混合物，4-mers占优势（75 +/- 0.4% w/w）。在21天的培养中，添加了纯HA 4-mers或低聚物混合物的细胞层的增殖水平与未添加HA的对照组相似。与对照相比，纯4-聚物和低聚物混合物分别使tropoelastin的dna归一化产量提高1.6倍和1.8倍，基质弹性蛋白的产量提高2.7倍。十二烷基硫酸钠(SDS)-PAGE/Western Blot和去氨酶半定量分析分别证实了观察到的对偶弹性蛋白和基质弹性蛋白的生化趋势。透明质酸低聚物诱导弹性蛋白交联剂桥氨的合成增强，并通过抑制弹性蛋白层粘连蛋白受体（ELR）活性来稳定弹性蛋白基质。透射电子显微镜（tem）显示，在低聚物补充的培养物中，弹性蛋白沉积明显，纵向定向，聚集原纤维和团块，而在对照组中，弹性蛋白沉积较少，且大部分为无定形。透明质酸低聚物保留了正常的纤维蛋白介导的弹性蛋白基质沉积。结果表明，透明质酸寡聚物具有高度的促弹性性，促进弹性蛋白纤维的形成，并稳定弹性蛋白基质，因此可以有效地加入到引导弹性蛋白再生的支架中。(c) 2006 Elsevier Ltd.版权所有。
Prior studies suggest that hyaluronan (HA), a glycosaminoglycan, may upregulate innately poor elastin matrix synthesis by adult vascular smooth muscle cells (SMCs). HA scaffolds could thus be useful to regenerate damaged vascular elastin. In an earlier study, we established that the elastogenic effects of non-oligomeric HA are fragment size- and/or dose-specific. We currently investigate the pro-elastogenic effects of exogenous HA oligomers on rat aortic smooth muscle cells (RASMCs). RASMCs were cultured with pure HA oligomers (4-mers) and mixtures (4-8 mers) obtained by enzymatic digestion of long-chain HA (MW similar to 2000 kDa). Polyacrylamide gel electrophoresis (PAGE)/Matrix Assisted Laser Desorption/Ionization Spectroscopy Time-Of-Flight Analysis (MALDI-TOF) showed HA digestates to contain a mixture of 4-8 mers with a predominance of 4-mers (75 +/- 0.4% w/w). Cell layers supplemented with both pure HA 4-mers or oligomer mixtures showed proliferation levels similar to non-HA controls over 21 days of culture. Pure 4-mers and oligomer mixtures enhanced DNA-normalized output of tropoelastin by 1.6 and 1.8 times, respectively, and that of matrix elastin by similar to 2.7 times relative to controls. Sodium dodecyl sulfate (SDS)-PAGE/Western Blot and a desmosine assay semi-quantitatively confirmed the observed biochemical trends for tropoelastin and matrix elastin, respectively. HA oligomers induced enhanced synthesis of the elastin crosslinker, desmosine, and appeared to stabilize the elastin matrix by suppression of elastin-laminin receptor (ELR) activity relative to controls. Transmission electron micrographs (TEMs) showed elastin deposits within oligomer-supplemented cultures to be distinct, longitudinally oriented, aggregating fibrils, and clumps, and to be less abundant and mostly amorphous in controls. HA oligomers preserved normal fibrillin-mediated elastin matrix deposition. Results suggest that HA oligos are highly pro-elastogenic, promote elastin fibril formation, and stabilize elastin matrix and may thus be usefully incorporated into scaffolds for guided elastin regeneration. (c) 2006 Elsevier Ltd. All rights reserved.