Different responses of galanin and calcitonin gene-related peptide to capsaicin stimulation on dorsal root ganglion neurons in vitro

Yunfeng Li , Guixiang Liu , Hao Li , Huaijing Wang , Zhen Liu
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引用次数: 3

Abstract

Both galanin (Gal) and calcitonin gene-related peptide (CGRP) are sensory neuropeptides which expressed in dorsal root ganglion (DRG) neurons and are involved in nociceptive processing. Capsaicin (CAP) influences nociceptive processing via influencing the expression of sensory neuropeptides in primary sensory neurons. However, little is known about the alterations of Gal and CGRP expression at the same condition stimulated by CAP. In the present study, primary cultured DRG neurons were used to determine the different responses of Gal and CGRP to CAP stimulation. DRG neurons were cultured for 48 hours and then exposed to CAP (2 μmol/L), capsazepine (CPZ) (2 μmol/L) plus CAP (2 μmol/L), or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 (10 μmol/L) plus CAP (2 μmol/L) for an additional 24 hours. The DRG neurons were continuously exposed to culture media as a control. After that, the levels of Gal mRNA and CGRP mRNA of DRG neurons were determined using real time-PCR analysis. Gal and CGRP expression in situ was detected by an immunofluorescent labeling technique. The levels of phosphorylated-ERK1/2 (pERK1/2) protein were detected using a Western blot assay. The results showed that CAP evoked increases of Gal and its mRNA and decreases of CGRP and its mRNA in DRG neurons. Administration of either CPZ or PD98059 blocked the effects of CAP. These data indicate that Gal and CGRP shared different responses to CAP stimulation. Gal and CGRP may have different effects in nociceptive processing during neurogenic inflammation.

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甘丙氨酸和降钙素基因相关肽对辣椒素刺激背根神经节神经元的不同反应
甘丙肽(Gal)和降钙素基因相关肽(CGRP)都是表达于背根神经节(DRG)神经元的感觉神经肽,参与伤害性加工。辣椒素(CAP)通过影响初级感觉神经元中感觉神经肽的表达来影响伤害性加工。然而,在CAP刺激的相同条件下,Gal和CGRP的表达变化知之甚少。本研究采用原代培养的DRG神经元来确定Gal和CGRP对CAP刺激的不同反应。将DRG神经元培养48 h后,分别暴露于CAP (2 μmol/L)、capsazepine (2 μmol/L) + CAP (2 μmol/L)或胞外信号调节激酶1/2 (ERK1/2)抑制剂PD98059 (10 μmol/L) + CAP (2 μmol/L)中24 h。作为对照,DRG神经元连续暴露于培养基中。real - time-PCR检测DRG神经元Gal mRNA和CGRP mRNA水平。采用免疫荧光标记技术检测原位Gal和CGRP的表达。Western blot法检测磷酸化erk1 /2 (pERK1/2)蛋白水平。结果表明,CAP诱导DRG神经元中Gal及其mRNA表达升高,CGRP及其mRNA表达降低。CPZ或PD98059均可阻断CAP的作用。这些数据表明Gal和CGRP对CAP刺激有不同的反应。Gal和CGRP可能在神经源性炎症的伤害加工中有不同的作用。
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Regulatory Peptides
Regulatory Peptides 医学-内分泌学与代谢
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2 months
期刊介绍: Regulatory Peptides provides a medium for the rapid publication of interdisciplinary studies on the physiology and pathology of peptides of the gut, endocrine and nervous systems which regulate cell or tissue function. Articles emphasizing these objectives may be based on either fundamental or clinical observations obtained through the disciplines of morphology, cytochemistry, biochemistry, physiology, pathology, pharmacology or psychology.
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