Effect and mechanism of the CACNA2D1-CGRP pathway in osteoarthritis-induced ongoing pain
Effect and mechanism of the CACNA2D1-CGRP pathway in osteoarthritis-induced ongoing pain
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CACNA2D1-CGRP通路在骨关节炎所致持续性疼痛中的作用及机制
DOI：
10.1016/j.biopha.2020.110374
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发表时间：
2020-09-01
影响因子：
7.5
通讯作者：
Liu，Ping
中科院分区：
文献类型：
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作者：
Sun，Liang;Wang，Guodong;Liu，Ping
本研究通过注射碘乙酸钠（MIA）建立大鼠骨关节炎（OA）模型，探讨电压依赖性钙通道亚单位α 2/δ 1（CACNA 2D 1）-降钙素基因相关蛋白（CGRP）通路在OA持续性疼痛中的作用及机制。CACNA 2D 1表达通过qPCR测定、蛋白质印迹测定和免疫荧光测量。采用热缩足潜伏期（PWL）和机械缩足阈（PWT）评价大鼠的疼痛行为。检测CACNA 2D 1、神经肽Y（NPY）、转录激活因子3（ATF 3）、CGRP、蛋白激酶A（PKA）、磷酸化（p）-PKA、腺苷酸环化酶（AC）、蛋白激酶C（PKC）、p-PKC、磷脂酶C（PLC）、丝裂原活化蛋白激酶（MAPK）信号通路蛋白的表达。CACNA 2D 1的敲低导致OA大鼠痛阈升高，而CACNA 2D 1的过表达则降低正常大鼠痛阈。CACNA 2D 1过表达抑制CGRP的表达，上调NPY、ATF 3、p-PKA、AC、p-PKC、PLC、p-Jun N末端激酶（JNK）和p-p38的表达，对磷酸化细胞外信号调节激酶（p-ERK）的表达无明显影响。利用MIA诱导的OA模型，我们证明CACNA 2D 1可能通过调节背根神经节中的CGRP和AC-PKA/PKC/MAPK信号通路参与疼痛过程。
This study built an OA model in rats by monosodium iodoacetate (MIA) injection to determine the effects and mechanism of the voltage-dependent calcium channel subunit alpha-2/delta-1 (CACNA2D1)-calcitonin gene-related protein (CGRP) pathway in osteoarthritis (OA)-induced ongoing pain. CACNA2D1 expression was measured by qPCR assay, western blotting assay, and immunofluorescence. Pain behaviors in rats were assessed with the measurement of thermal paw withdrawal latency (PWL) and mechanical paw withdrawal threshold (PWT). The expression of CACNA2D1, neuropeptide Y (NPY), activating transcription factor 3 (ATF3), CGRP, protein kinase A (PKA), phosphorylated (p)-PKA, adenylyl cyclase (AC), protein kinase C (PKC), p-PKC, phospholipase C (PLC), and mitogen-activated protein kinase (MAPK) signaling pathway proteins were measured, OA rats had higher CACNA2D1 expression than normal rats. Knockdown of CACNA2D1 led to the elevation of the pain threshold of OA rats, and CACNA2D1 over-expression decreased the pain threshold of normal rats. Moreover, CACNA2D1 over-expression inhibited the expression of CGRP, up-regulated the expressions of NPY, ATF3, p-PKA, AC, p-PKC, PLC, p-Jun N-terminal kinase (JNK), and p-p38, and had no significant effect on phosphorylated extracellular signal-regulated kinase (p-ERK) expressionin vivoandin vitro. Using this model of MIA-induced OA, we demonstrated that CACNA2D1 might be involved in the process of pain by modulating the CGRP and AC-PKA/PKC/MAPK signaling pathways in the dorsal root ganglion.