HUMAN PRIMARY VASCULAR SMOOTH MUSCLE CELL CULTURE

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们认为，铁在心血管疾病中起着关键作用：血管平滑肌细胞（VSMC）的迁移和增殖。为了验证这一假设，我们将使用我们描述和合成的一个特殊的铁螯合剂家族。这个家族是外螯素，由结核分枝杆菌分泌的铁载体。它们阻断高毒性羟基自由基（oOH）的铁依赖性产生。外螯素是独特的，因为它们是脂溶性的，我们假设的一种性质使它们能够迅速进入细胞并定位在细胞膜的脂质部分，在那里发生关键的铁介导的过程。 脱铁-外螯素（D-Exo）也阻止培养的血管平滑肌细胞的生长。我们将采用exochelins深入了解铁依赖性机制，导致血管平滑肌生长。铁是细胞中普遍存在的金属，存在于许多酶和蛋白质中。细胞增殖所必需的分子之一是核糖核苷酸还原酶（RR），一种需要铁的酶。RR是将核糖核苷酸还原为脱氧核糖核苷酸所必需的，这是DNA合成的关键步骤。我们对D-Exo处理的细胞进行了微阵列分析，发现一种持续上调的分子是过氧化物酶体增殖物激活受体-γ。已经显示，PPAR配体通过干扰血管和癌细胞中多种细胞周期调节剂的表达和功能来抑制这些细胞的生长。我们建议测试的假设，D-Exo抑制人血管平滑肌细胞增殖，抑制细胞周期退出从G1期进入S期，通过减弱Rb磷酸化，通过过氧化物酶体增殖受体。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We propose that iron plays a critical role in cardiovascular diseases: vascular smooth muscle cell (VSMC) migration and proliferation. To test this hypothosis we will employ an extraordinary family of iron chelators that we have described and synthesized. This family is exochelins, siderophores secreted by Mycobacterium tuberculosis. They block the iron-dependent production of highly toxic hydroxyl radical (oOH). Exochelins are unique in that they are lipid soluble, a property that we postulate enables them to rapidly enter cells and localize in the lipid portion of the cell membrane where critical iron-mediated processes occur. Desferri-exochelins (D-Exo) also prevent growth of cultured vascular smooth muscle cells. We will employ exochelins to gain insights into the iron-dependent mechanisms leading to vascular smooth muscle growth. Iron is a ubiquitous metal in cells and is present in many enzymes and proteins. One of the molecules necessary for cell proliferation is ribonucleotide reductase (RR), an iron requiring enzyme. RR is indispensable for the reduction of ribonucleotides to deoxyribonulceotides, a critical step in DNA synthesis. we have done micro array analysis on D-Exo treated cells and found that one molecule that is consistently up-regulated is peroxisome proliferator-activated receptor-gamma. PPAR ligands have been shown to inhibit growth of vascular and cancer cells by interfering with the expression and function of multiple cell cycle regulators in these cells. We propose to test the hypothesis that D-Exo inhibits human vascular smooth muscle cell proliferation by inhibiting the cell cycle exit from G1 into S phase by attenuating Rb phosphorylation through PPAR.