Herembert T, Gogusev J, Zhu D L, Drueke T B, Marche P
Université René Descartes and Department of Pharmacology, CNRS URA 1482, CHU Necker, 156 rue de Vaugirard, 75015 Paris, France.
Biochem J. 1997 Jul 1;325 ( Pt 1)(Pt 1):123-8. doi: 10.1042/bj3250123.
Since in several vascular diseases abnormal vascular smooth-muscle cell (VSMC) proliferation is often associated with the presence of macrophages, we examined whether macrophage-colony-stimulating factor (M-CSF) might play a role in the control of VSMC growth. VSMCs were isolated from rat aorta and maintained in culture. Using a bioassay, a macrophage-colony-stimulating activity was detected in the serum-free supernatant of VSMCs, which could be inhibited by the addition of specific anti-M-CSF antibodies. The presence of M-CSF receptor protein and of M-CSF and M-CSF receptor gene transcripts was demonstrated by immunocytochemistry, using a specific anti-c-Fms antibody and Northern blot analysis respectively. [3H]Thymidine incorporation was measured following the addition to quiescent VSMCs of various dilutions of L929 cell supernatant (as a source of M-CSF) or of recombinant M-CSF. Both exogenous M-CSF and serum-free VSMC conditioned medium promoted DNA synthesis in a concentration-dependent manner, and this effect could be abrogated by the presence of a specific anti-M-CSF antibody. Under similar experimental conditions, L929 cell supernatant modulated proto-oncogene expression, as assessed by Northern blot analysis of c-fos, c-myc, egr-1 and junB. It was further demonstrated that M-CSF could act in synergy with thrombin, platelet-derived growth factor or basic fibroblast growth factor in promoting VSMC DNA synthesis. These results support the hypothesis that M-CSF affects the growth of cultured rat VSMCs through paracrine/autocrine mechanisms. Its effects at both the macrophage and the VSMC level confer to M-CSF a central role in the development of vascular lesions that occurs during atherosclerotic progression.
由于在几种血管疾病中，异常的血管平滑肌细胞（VSMC）增殖常与巨噬细胞的存在有关，我们研究了巨噬细胞集落刺激因子（M-CSF）是否可能在VSMC生长控制中发挥作用。从大鼠主动脉分离VSMC并进行培养。通过生物测定法，在VSMC的无血清上清液中检测到巨噬细胞集落刺激活性，加入特异性抗M-CSF抗体可抑制该活性。分别使用特异性抗c-Fms抗体和Northern印迹分析，通过免疫细胞化学证明了M-CSF受体蛋白以及M-CSF和M-CSF受体基因转录本的存在。在静止的VSMC中加入各种稀释度的L929细胞上清液（作为M-CSF的来源）或重组M-CSF后，测量[3H]胸腺嘧啶掺入量。外源性M-CSF和无血清VSMC条件培养基均以浓度依赖性方式促进DNA合成，并且特异性抗M-CSF抗体的存在可消除这种作用。在类似的实验条件下，通过对c-fos、c-myc、egr-1和junB的Northern印迹分析评估，L929细胞上清液调节原癌基因表达。进一步证明，M-CSF在促进VSMC DNA合成方面可与凝血酶、血小板衍生生长因子或碱性成纤维细胞生长因子协同作用。这些结果支持以下假设：M-CSF通过旁分泌/自分泌机制影响培养的大鼠VSMC的生长。其在巨噬细胞和VSMC水平上的作用使M-CSF在动脉粥样硬化进展过程中发生的血管病变发展中起核心作用。
