AUTOCRINE AND PARACRINE EFFECTS OF ATRIAL-NATRIURETIC-PEPTIDE GENE-TRANSFER ON VASCULAR SMOOTH-MUSCLE AND ENDOTHELIAL CELLULAR GROWTH
AUTOCRINE AND PARACRINE EFFECTS OF ATRIAL-NATRIURETIC-PEPTIDE GENE-TRANSFER ON VASCULAR SMOOTH-MUSCLE AND ENDOTHELIAL CELLULAR GROWTH
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DOI：
10.1172/jci117402
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发表时间：
1994-08-01
影响因子：
15.9
通讯作者：
DZAU, VJ
中科院分区：
文献类型：
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作者：
MORISHITA, R;GIBBONS, GH;DZAU, VJ
除了心房，最近的证据表明心钠素(ANP)也在其他组织中合成。特别令人感兴趣的是ANP mRNA在血管壁中的位置。我们和其他人已经证明，外源性添加AI？IP抑制培养的内皮细胞和血管平滑肌细胞(VSMC)的生长。然而，目前还不知道本地合成的ANP是否会有类似的作用。由于培养的内皮细胞和VSMC已经失去了表达内源性ANP基因的能力，我们用表达大鼠ANP的表达载体转染培养的细胞，并检测了其对细胞生长的影响。在基础和成纤维细胞生长因子刺激条件下，这些细胞的DNA合成率也显著降低。将ANP载体转染内皮细胞的条件培养液加入到未转染内皮细胞中，可显著增加细胞内cGMP的表达。同样，从血管内皮细胞收集的ANP载体条件培养液也减少了VSMC的DNA合成。转染ANP的内皮细胞与静止的VSMC共培养发现，内皮细胞释放的ANP抑制VSMC的DNA合成。最后，FOE检测了ANP载体直接导入VSMC的自分泌效应。在基础和血管紧张素II刺激条件下，ANP载体的转染会降低VSMC的DNA合成。同样，转染ANP载体后，PDGF和血清(5%)刺激的VSMC DNA合成减少。这些结果表明，内源性ANP对血管内皮细胞和VSMC的生长具有自分泌和旁分泌抑制作用。体内ANP基因转移可能为血管收缩和病理性生长异常的血管疾病提供基因治疗的机会。
In addition to the atria, recent evidence suggests that atrial natriuretic peptide (ANP) is also synthesized in other tissues. Of particular interest is the location of ANP mRNA in the vessel wall. We and others have shown that exogenously added AI?IP inhibited the growth of endothelial cells and vascular smooth muscle cells (VSMC) in culture. However, it is not known if the locally synthesized ANP would act similarly. Because cultured endothelial cells and VSMC have lost the ability to express the endogenous ANP gene, we have transfected cells in culture with an expression vector expressing rat ANP and have examined the effects on growth.Cultured endothelial cells transfected with an ANP expression vector synthesized and secreted high levels of ANP. These cells also showed significantly lower rates of DNA synthesis under basal and fibroblast growth factor (FGF)stimulated conditions. Addition of conditioned medium from endothelial cells transfected with ANP vector to nontransfected endothelial cells resulted in the significant increase in cyclic GMP. Similarly, conditioned media collected from endothelial cells transfected with ANP vector also decreased DNA synthesis in VSMC. Coculture of ANP-transfected endothelial cells with quiescent VSMC showed that released ANP from endothelial cells inhibited DNA synthesis in VSMC. Finally, Foe examined the autocrine effect of direct transfection of ANP vector into VSMC. Transfection of the ANP vector decreased DNA synthesis in VSMC under basal and angiotensin II-stimulated conditions. Similarly, transfection of the ANP vector resulted in a decrease in the PDGF and serum (5%)-stimulated DNA synthesis of VSMC. These results demonstrate that endogenously produced ANP can exert autocrine and paracrine inhibitory effects on endothelial cell and VSMC growth. In vivo gene transfer of ANP may provide us with the opportunity of gene therapy for vascular diseases in which the abnormalities are vasoconstriction and pathological growth.