Keller H U, Niggli V
Institute of Pathology, University of Bern, Switzerland.
J Cell Physiol. 1994 Dec;161(3):526-36. doi: 10.1002/jcp.1041610316.
Using two newly synthesized inhibitors, Ro 31-8220 and CGP 41,251, of protein kinase C (PKC), we analysed: (1) how distinct PMN functions (shape changes, locomotion, pinocytosis) are regulated, and (2) the role of protein phosphorylation and PKC in this process. We were able to transform: (1) resting PMNs into locomoting cells using fNLPNTL, (2) locomoting cells into non-locomoting highly pinocytic cells using PMA, and (3) PMA-stimulated cells showing marked pinocytosis into locomoting or into resting cells using Ro 31-8220. It is thus possible to selectively manipulate PMN function (resting state, locomotion, marked pinocytosis), indicating that there are different regulatory pathways. It was not possible to induce locomotion and marked pinocytosis simultaneously, indicating crosstalk between pathways. Ro 31-8220 inhibited PMA-induced shape changes (nonpolar cells) and pinocytosis, but not fNLPNTL-induced shape changes (polarity) and pinocytosis. At higher concentrations, Ro 31-8220 alone elicited cell polarity and chemokinesis, indicating that a constitutively active protein kinase is involved in maintaining the spherical shape of resting PMNs. Functional effects of another PKC inhibitor, CGP 41,251, on neutrophil function were strikingly different. CGP 41,251 selectively inhibited fNLPNTL-induced polarity and locomotion (but not colchicine or Ro 31-8220-induced polarity), and it failed to inhibit PMA-induced, stimulated pinocytosis and shape changes. Although the effects of Ro 31-8220 vs. CGP 41,251 on PMN function were strikingly different, the inhibition of profiles for constitutive and for fNLPNTL- or PMA-induced protein phosphorylation in intact PMNs showed only small differences, which could not yet be conclusively related to cell function.
我们使用两种新合成的蛋白激酶C（PKC）抑制剂Ro 31-8220和CGP 41,251，分析了：（1）不同的中性粒细胞功能（形态变化、运动、胞饮作用）是如何被调节的，以及（2）蛋白质磷酸化和PKC在此过程中的作用。我们能够实现：（1）使用fNLPNTL将静息中性粒细胞转化为运动细胞，（2）使用佛波酯（PMA）将运动细胞转化为非运动的高胞饮细胞，以及（3）使用Ro 31-8220将显示明显胞饮作用的PMA刺激细胞转化为运动细胞或静息细胞。因此，有可能选择性地操控中性粒细胞功能（静息状态、运动、明显胞饮作用），这表明存在不同的调节途径。不可能同时诱导运动和明显胞饮作用这表明途径之间存在相互作用。Ro 31-8220抑制PMA诱导的形态变化（非极性细胞）和胞饮作用，但不抑制fNLPNTL诱导的形态变化（极性）和胞饮作用。在较高浓度下，单独的Ro 31-8220可引发细胞极性和化学趋化作用，表明一种组成型活性蛋白激酶参与维持静息中性粒细胞的球形形态。另一种PKC抑制剂CGP 41,251对中性粒细胞功能的影响则显著不同。CGP 41,251选择性抑制fNLPNTL诱导的极性和运动（但不抑制秋水仙碱或Ro 31-8220诱导的极性），并且它未能抑制PMA诱导的刺激胞饮作用和形态变化。尽管Ro 31-8220与CGP 41,251对中性粒细胞功能的影响显著不同，但在完整中性粒细胞中，对组成型以及fNLPNTL或PMA诱导的蛋白质磷酸化的抑制谱仅显示出微小差异，这些差异尚未能与细胞功能明确关联。
