Peptide Analysis In Neuronal Cell Culture

神经肽及其类似物通常在极低的浓度下工作，需要开发极其灵敏的技术来分析它们。为探讨血管活性肠肽(VIP)和垂体腺苷环化酶激活肽(PACAP)对星形胶质细胞培养的影响，建立了免疫亲和毛细管电泳法(ICE)。该程序包括在分析来自星形胶质细胞培养的条件培养液之前，将抗体固定在直径100微米的熔融石英毛细血管内表面。ICE系统能够分离和测量刺激细胞分泌的15种不同的细胞因子和趋化因子。研究表明，用皮摩尔到飞摩尔浓度的神经肽刺激星形胶质细胞，可以诱导大量炎性细胞因子和趋化因子的产生和分泌。这些发现表明某些神经肽在局部炎症过程中可能发挥作用。

Neuropeptides and their analogs often work at extremely low concentrations requiring the development of extremely sensitive techniques for their analysis. To examine the effects of the peptides Vasoactive Intestinal Peptide (VIP) and Pituitary Adenylate Cyclase Activating Peptide (PACAP) on astrocyte cultures, an immunoaffinity capillary electrophoresis (ICE) technique was developed. The procedure involves immobilization of antibodies to the internal surface of a 100-micron diameter fused silica capillary prior to analysis of the conditioned medium derived from the astrocyte cultures. The ICE system was able to isolate and measure 15 different cytokines and chemokines secreted by the stimulated cells. The studies demonstrate that stimulation of astrocytes with pico- to femto-molar concentrations of the neuropeptides induces the production and secretion of a number of inflammatory cytokines and chemokines. These findings indicate a potential role for certain neuropeptides in the local inflammatory process.