Antisense oligonucleotides to stromelysin mRNA inhibit injury-induced proliferation of arterial smooth muscle cells
Antisense oligonucleotides to stromelysin mRNA inhibit injury-induced proliferation of arterial smooth muscle cells
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DOI：
10.14670/hh-14.1101
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发表时间：
1999-10-01
影响因子：
2
通讯作者：
Hultgårdh-Nilsson, A
中科院分区：
文献类型：
--
作者：
Lövdahl, C;Thyberg, J;Hultgårdh-Nilsson, A
平滑肌细胞迁移和增殖是动脉粥样硬化和球囊血管成形术后再狭窄相关内膜病变形成的重要事件。为了使这成为可能，平滑肌细胞必须从收缩细胞转变为具有合成DNA和细胞外基质成分的能力的活化修复细胞。现在有相当多的证据表明，细胞外基质在调节平滑肌细胞的表型特性中具有重要的功能，但对基质金属蛋白酶的作用知之甚少。本研究探讨了基质溶解素在大鼠主动脉平滑肌细胞形态和功能的调制在体外和体内机械损伤后的作用。反义mRNA寡核苷酸被用来研究基质分解素表达在损伤诱导的表型调节和随后的血管平滑肌细胞的迁移和增殖中的作用。以培养的大鼠主动脉平滑肌细胞和球囊损伤的大鼠颈动脉作为实验模型。光镜和电镜观察平滑肌细胞表型和病变形成的变化，并掺入H-3-胸苷检测DNA合成。损伤诱导的DNA合成和迁移在体外被抑制72%和36%，分别通过在损伤前向培养基中加入基质分解素反义寡核苷酸。在原代培养物中，67%的平滑肌细胞与基质分解素反义处理保留在收缩表型，通过细胞精细结构分析判断，相比之下，15%未处理的细胞和40%的细胞与错配的寡核苷酸处理。球囊损伤后一周的颈动脉检查同样表明，与用错配寡核苷酸处理的那些相比，用反义寡核苷酸处理的血管中的中膜的内部部分中的收缩细胞的比例更大。此时，反义处理动脉的新生内膜也明显薄于错配处理动脉和对照动脉。这些发现表明，基质分解素mRNA反义寡核苷酸抑制大鼠动脉平滑肌细胞的表型调节，从而导致血管壁损伤引起的迁移和增殖以及新生内膜形成的减少。
Smooth muscle cell migration and proliferation are important events in the formation of intimal lesions associated with atherosclerosis and restenosis following balloon angioplasty. To make this possible, the smooth muscle cell has to change from a contractile to an activated repair cell with capacity to synthesize DNA and extracellular matrix components. There is now considerable evidence that the extracellular matrix has important functions in modulating the phenotypic properties of smooth muscle cells, but less is known about the role of the matrix metalloproteinases. The present study investigates the role of stromelysin in the modulation of rat aortic smooth muscle cell morphology and function following mechanical injury in vitro and in vivo. Antisense mRNA oligonucleotides were used to investigate the role of stromelysin expression in injury-induced phenotypic modulation and the subsequent migration and proliferation of vascular smooth muscle cells. Cultured rat aortic smooth muscle cells and balloon-injured rat carotid arteries were used as experimental models. Light- and electron microscopy were used to follow changes in smooth muscle cell phenotype and lesion formation and incorporation of H-3-thymidine to detect DNA synthesis. Injury-induced DNA synthesis and migration in vitro were inhibited by 72% and 36%, respectively, by adding stromelysin antisense oligonucleotides to the medium prior to injury. In primary cultures, 67% of the smooth muscle cells treated with stromelysin antisense were retained in a contractile phenotype as judged by analysis of cell fine structure, compared to 15% untreated cells and 40% in cells treated with mismatched oligonucleotides. Examination of the carotid arteries one week after balloon injury likewise demonstrated a larger fraction of contractile cells in the inner parts of the media in vessels treated with antisense oligonucleotides compared to those treated with mismatched oligonucleotides. The neointima was also distinctly thinner in antisense-treated than in mismatched-treated and control arteries at this time. These findings indicate that stromelysin mRNA antisense oligonucleotides inhibited phenotypic modulation of rat arterial smooth muscle cells and so caused a decrease in migration and proliferation and neointima formation in response to vessel wall injury.