Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2b) mediates oxidation-induced endoplasmic reticulum stress to regulate neuropathic pain
Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2b) mediates oxidation-induced endoplasmic reticulum stress to regulate neuropathic pain
复制标题
肌浆/内质网 Ca2-ATP 酶 (SERCA2b) 介导氧化诱导的内质网应激以调节神经性疼痛
DOI：
10.1111/bph.15744
复制
发表时间：
2022-01-13
影响因子：
7.3
通讯作者：
Cao, Zhengyu
中科院分区：
文献类型：
--
作者：
Li, Shaoheng;Zhao, Fang;Cao, Zhengyu
背景和目的神经病理性疼痛是一种广泛存在的健康问题，但治疗方法有限。肌浆网/内质网Ca ~（2+）-ATP酶（SERCA）在神经病理性疼痛动物的背根神经节（DRG）中表达降低。我们的目的是建立SERCA表达和疼痛反应之间的关系，并阐明潜在的分子机制。实验方法使用大鼠慢性压迫性损伤（CCI）建立神经病理性疼痛模型。采用钙离子成像和电流钳膜片钳分别测定细胞内钙离子水平和动作电位放电。Western印迹、免疫荧光染色和qRT-PCR分别用于定量评估蛋白质和mRNA表达。分别采用H&E染色和偶联酶测定来评价神经损伤和SERCA 2b活性。关键结果SERCA 2b是大鼠DRG中主要的SERCA亚型，CCI后其表达在mRNA、蛋白质和活性水平上降低。然而，用毒胡萝卜素抑制SERCA会导致神经元过度兴奋、神经损伤、内质网（ER）应激、卫星神经胶质细胞活化和机械异常性疼痛，CCI后通过CDN 1163激活SERCA或在DRG中过表达SERCA 2b产生机械和热异常性疼痛的长期缓解，伴随着通过缓解ER应激的形态和功能恢复。此外，CCI大鼠DRG SERCA 2b的下调是由通过Sp1依赖性转录抑制增加ROS产生引起的。结论和意义我们的研究结果揭示了一个新的途径为中心的SERCA 2b的发展和维持的神经病理性疼痛的机制的关键分子，和SERCA 2b激活剂具有潜在的治疗性治疗神经病理性疼痛。
Background and Purpose Neuropathic pain is a widespread health problem with limited curative treatment. Decreased sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) expression has been reported in dorsal root ganglion (DRG) of animals suffering from neuropathic pain. We aimed to establish the relationship between SERCA expression and the pain responses and to elucidate the underlying molecular mechanism. Experimental Approach Neuropathic pain was modelled using rat chronic constriction injury (CCI). Ca2+ imaging and current clamp patch-clamp were used to determine cytosolic Ca2+ levels and action potential firing, respectively. Western blots, immunofluorescence staining and qRT-PCR were used to quantitatively assess protein and mRNA expression, respectively. H&E staining and coupled enzyme assays were used to evaluate the nerve injury and SERCA2b activity, respectively. Key Results SERCA2b is the predominant SERCA isoform in rat DRG and its expression is decreased after CCI at mRNA, protein and activity levels. Whereas inhibiting SERCA with thapsigargin causes neuronal hyperexcitation, nerve injury, endoplasmic reticulum (ER) stress, satellite glial cell activation and mechanical allodynia, activating SERCA by CDN1163 or overexpressing SERCA2b in DRG after CCI produces long-term relief of mechanical and thermal allodynia accompanied by morphological and functional restoration through alleviation of ER stress. Furthermore, the down-regulation of DRG SERCA2b in CCI rats is caused by increased production of ROS through Sp1-dependent transcriptional inhibition. Conclusion and Implications Our findings reveal a novel pathway centring around SERCA2b as the key molecule underlying the mechanism of development and maintenance of neuropathic pain, and SERCA2b activators have the potential for therapeutic treatment of neuropathic pain.