Lin You-Min, Fu Yu, Winston John, Radhakrishnan Ravi, Sarna Sushil K, Huang Li-Yen M, Shi Xuan-Zheng
Departments of Internal Medicine.
Surgery and.
Pain. 2017 Apr;158(4):583-592. doi: 10.1097/j.pain.0000000000000797.
Abdominal pain is one of the major symptoms in bowel obstruction (BO); its cellular mechanisms remain incompletely understood. We tested the hypothesis that mechanical stress in obstruction upregulates expression of nociception mediator nerve growth factor (NGF) in gut smooth muscle cells (SMCs), and NGF sensitizes primary sensory nerve to contribute to pain in BO. Partial colon obstruction was induced with a silicon band implanted in the distal bowel of Sprague-Dawley rats. Colon-projecting sensory neurons in the dorsal root ganglia (T13 to L2) were identified for patch-clamp and gene expression studies. Referred visceral sensitivity was assessed by measuring withdrawal response to stimulation by von Frey filaments in the lower abdomen. Membrane excitability of colon-projecting dorsal root ganglia neurons was significantly enhanced, and the withdrawal response to von Frey filament stimulation markedly increased in BO rats. The expression of NGF mRNA and protein was increased in a time-dependent manner (day 1-day 7) in colonic SMC but not in mucosa/submucosa of the obstructed colon. Mechanical stretch in vitro caused robust NGF mRNA and protein expression in colonic SMC. Treatment with anti-NGF antibody attenuated colon neuron hyperexcitability and referred hypersensitivity in BO rats. Obstruction led to significant increases of tetrodotoxin-resistant Na currents and mRNA expression of Nav1.8 but not Nav1.6 and Nav1.7 in colon neurons; these changes were abolished by anti-NGF treatment. In conclusion, mechanical stress-induced upregulation of NGF in colon SMC underlies the visceral hypersensitivity in BO through increased gene expression and activity of tetrodotoxin-resistant Na channels in sensory neurons.
腹痛是肠梗阻(BO)的主要症状之一;其细胞机制仍未完全明确。我们验证了以下假说:梗阻中的机械应力会上调肠道平滑肌细胞(SMC)中伤害感受介质神经生长因子(NGF)的表达,且NGF使初级感觉神经敏感化,从而导致BO疼痛。在Sprague-Dawley大鼠的远端肠段植入硅胶带诱导部分结肠梗阻。鉴定出背根神经节(T13至L2)中投射至结肠的感觉神经元,用于膜片钳和基因表达研究。通过测量对下腹von Frey细丝刺激的退缩反应来评估牵涉性内脏敏感性。在BO大鼠中,投射至结肠的背根神经节神经元的膜兴奋性显著增强,对von Frey细丝刺激的退缩反应明显增加。结肠SMC中NGF mRNA和蛋白的表达在第1天至第7天呈时间依赖性增加,但在梗阻结肠的黏膜/黏膜下层中未增加。体外机械牵张可导致结肠SMC中NGF mRNA和蛋白的强烈表达。用抗NGF抗体治疗可减轻BO大鼠结肠神经元的过度兴奋和牵涉性超敏反应。梗阻导致结肠神经元中河豚毒素抗性钠电流和Nav1.8的mRNA表达显著增加,但Nav1.6和Nav1.7未增加;抗NGF治疗可消除这些变化。总之,结肠SMC中机械应力诱导的NGF上调通过感觉神经元中基因表达增加和河豚毒素抗性钠通道活性增加,成为BO内脏超敏反应的基础。