He Yanling, Ding Guifeng, Xia Donglan, Zhu Tiejun, Fan Shaoguang
Department of Dermatology, People's Hospital, Skin and STD Research Center, Peking University, Beijing 100044, China.
Zhonghua Yi Xue Za Zhi. 2002 Jan 25;82(2):131-4.
To study the effect of calcitonin gene-related peptide (CGRP) on the function of immune cells.
Human monocytes were cultured with calcitonin gene-related peptide in vitro and activated with lipopolysaccharide (LPS). The IL-8 level in the supernatant was measured with ELISA and the IL-8 mRNA expression in monocytes was observed by reverse transcription polymerase chain reaction (RT-PCR). The chemotactic activity of monocytes to neutrophils and lymphocytes was analyzed with micro-chemotacxis chamber. Chemotactic index (CI) was calculated by the formula: number of monocytes migrating to the underside of membrane in the LPS + CGRP group/number of monocytes migrating to the underside of membrane in the control group. CGRP receptor antagonist CGRP8 - 37 was added into the culture to study the effect of CGRP. Blank control and cultures of monocytes with LPS or with CGRP only were used as controls.
The level of IL-8 protein in the supernatant of the LPS + CGRP group was 1 120 pg/ml +/- 14.80 pg/ml, significantly higher than those in other groups (670 pg/ml +/- 15.10 pg/ml in LPS + CGRP + CGRP8 - 37 group). The expression of IL-8 mRNA in the LPS + CGRP group was the highest (IL-8/beta-actin = 1.845 +/- 0.587), IL-8/beta-actin in the LPS + CGRP + CGRP8 - 37 group was1.339 +/- 0.434. The chemotactic activities of the monocytes to neutrophils and lymphocytes were enhanced in the LPS + CGRP group (CI = 3.78 +/- 0.08 to neutrophils and CI = 3.4 +/- 0.27 to lymphocytes). The CI values were 1.15 +/- 0.31 and 1.21 +/- 0.06 respectively in the LPS + CGRP + CGRP 8 - 37 group.
CGRP in the peripheral nerve ending induces monocytes to synthetize and secret chemotactic factor IL-8 and enhance the chemotactic activity of monocytes, thus promoting the directional migration and aggregation of neutrophils and lymphocytes to foci of inflammation.
研究降钙素基因相关肽（CGRP）对免疫细胞功能的影响。
体外培养人单核细胞，加入降钙素基因相关肽，并使用脂多糖（LPS）激活。采用酶联免疫吸附测定法（ELISA）检测上清液中白细胞介素-8（IL-8）水平，通过逆转录聚合酶链反应（RT-PCR）观察单核细胞中IL-8 mRNA表达。使用微量趋化室分析单核细胞对中性粒细胞和淋巴细胞的趋化活性。趋化指数（CI）通过以下公式计算：LPS + CGRP组迁移至膜下侧的单核细胞数量/对照组迁移至膜下侧的单核细胞数量。在培养物中加入CGRP受体拮抗剂CGRP8 - 37，以研究CGRP的作用。以空白对照以及仅用LPS或仅用CGRP培养单核细胞作为对照。
LPS + CGRP组上清液中IL-8蛋白水平为1 120 pg/ml ± 14.80 pg/ml，显著高于其他组（LPS + CGRP + CGRP8 - 37组为670 pg/ml ± 15.10 pg/ml）。LPS + CGRP组中IL-8 mRNA表达最高（IL-8/β-肌动蛋白 = 1.845 ± 0.587），LPS + CGRP + CGRP8 - 37组中IL-8/β-肌动蛋白为1.339 ± 0.434。LPS + CGRP组中单核细胞对中性粒细胞和淋巴细胞的趋化活性增强（对中性粒细胞的CI = 3.78 ± 0.08，对淋巴细胞的CI = 3.4 ± 0.27）。LPS + CGRP + CGRP 8 - 37组的CI值分别为1.15 ± 0.31和1.21 ± 0.06。
外周神经末梢中的CGRP诱导单核细胞合成并分泌趋化因子IL-8，增强单核细胞的趋化活性，从而促进中性粒细胞和淋巴细胞向炎症灶定向迁移和聚集。
