Protein kinase C-α upregulates sodium channel Nav1.9 in nociceptive dorsal root ganglion neurons in an inflammatory arthritis pain model of rat
Protein kinase C-α upregulates sodium channel Nav1.9 in nociceptive dorsal root ganglion neurons in an inflammatory arthritis pain model of rat
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DOI：
10.1002/jcb.29322
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发表时间：
2019-08-05
影响因子：
4
通讯作者：
Dong, Tieli
中科院分区：
文献类型：
--
作者：
Bai, Qian;Shao, Jinping;Dong, Tieli
以往的研究发现，Nav1.9和蛋白激酶C（PKC）的表达增加有助于在一些炎症性疼痛模型的疼痛超敏反应。本研究旨在观察PKC对类风湿关节炎（RA）疼痛模型背根神经节（DRG）Nav1.9表达的调节作用。通过关节内注射完全弗氏佐剂（CFA）建立大鼠慢性膝关节炎模型。伤害性行为，包括机械，冷，热痛觉过敏进行了检查。采用定量聚合酶链反应、Western blot和免疫荧光法检测DRG中Nav1.9和PKC α的表达。检测了PKC激活剂（佛波醇12-肉豆蔻酸酯13-乙酸酯[PMA]）和PKC抑制剂（GF-109203 X）对Nav1.9表达的体外和体内作用。此外，研究了PKC调节剂对伤害性行为的影响。CFA注射后3至14天观察到机械、热和冷敏感性增加。免疫荧光实验发现，RA大鼠背根节神经元中Nav1.9和PKC α的mRNA和蛋白表达水平明显升高，Nav1.9优先与IB 4 +DRG神经元共定位。在培养的DRG神经元中，PMA增加Nav1.9的表达，而GF-109203 X阻止PMA的作用。PMA增加了未处理大鼠的Nav1.9表达，而GF-109203 X降低了RA大鼠的Nav1.9表达。在幼稚大鼠，PMA引起机械和冷痛觉过敏。另一方面，GF-109203 X减弱RA疼痛模型中的机械和冷痛觉过敏。在CFA诱导的RA疼痛慢性膝关节炎模型中，DRG中的Nav1.9可能被PKC α上调，这有助于疼痛超敏反应。
Previous studies have found that increased expression of Nav1.9 and protein kinase C (PKC) contributes to pain hypersensitivity in a couple of inflammatory pain models. Here we want to observe if PKC can regulate the expression of Nav1.9 in dorsal root ganglion (DRG) in rheumatoid arthritis (RA) pain model. A chronic knee joint inflammation model was produced by intra-articular injection of the complete Freund's adjuvant (CFA) in rats. Nociceptive behaviors including mechanical, cold, and heat hyperalgesia were examined. The expression of Nav1.9 and PKC alpha in DRG was detected by a quantitative polymerase chain reaction, Western blot, and immunofluorescence. The in vitro and in vivo effects of a PKC activator (phorbol 12-myristate 13-acetate [PMA]) and a PKC inhibitor (GF-109203X) on the expression of Nav1.9 were examined. Moreover, the effects of PKC modulators on nociceptive behaviors were studied. Increased mechanical, heat, and cold sensitivity was observed 3 to 14 days after CFA injection. Parallel increases in messenger RNA and protein expression of Nav1.9 and PKC alpha were found. Immunofluorescence experiments found that Nav1.9 was preferentially colocalized with IB4+DRG neurons in RA rats. In cultured DRG neurons, PMA increased Nav1.9 expression while GF-109203X prevented the effect of PMA. PMA increased Nav1.9 expression in naive rats while GF-109203X decreased Nav1.9 expression in RA rats. In naive rats, PMA caused mechanical and cold hyperalgesia. On the other hand, GF-109203X attenuated mechanical and cold hyperalgesia in RA-pain model. Nav1.9 might be upregulated by PKC alpha in DRG, which contributes to pain hypersensitivity in CFA-induced chronic knee joint inflammation model of RA pain.