Cell Biology and Toxicology ( IF 5.3 ) Pub Date : 2021-07-20 , DOI: 10.1007/s10565-021-09631-y Gui-Zhou Li 1, 2 , Ya-Hui Hu 1 , Yi-Ni Lu 2 , Qing-Yan Yang 1, 2 , Di Fu 1, 2 , Feng Chen 1 , Yun-Man Li 2
|
Vincristine (VCR), an alkaloid isolated from vinca, is a commonly used chemotherapeutic drug. However, VCR therapy can lead to dose-dependent peripheral neurotoxicity, mainly manifesting as neuropathic pain, which is one of the dominant reasons for limiting its utility. Experimentally, we discovered that VCR-induced neuropathic pain (VINP) was accompanied by astrocyte activation; the upregulation of phospho-CaMKII (p-CaMKII), CaV3.2, and Connexin-43 (Cx43) expression; and the production and release of inflammatory cytokines and chemokines in the spinal cord. Similar situations were also observed in astrocyte cultures. Interestingly, these alterations were all reversed by intrathecal injection of KN-93 (a CaMKII inhibitor) or l-Ascorbic acid (a CaV3.2 inhibitor). In addition, KN-93 and l-Ascorbic acid inhibited the increase in [Ca2+]i associated with astrocyte activation. We also verified that knocking down or inhibiting Cx43 level via intrathecal injection of Cx43 siRNA or Gap27 (a Cx43 mimetic peptide) relieved pain hypersensitivity and reduced the release of inflammatory factors; however, they did not affect astrocyte activation or p-CaMKII and CaV3.2 expression. Besides, the overexpression of Cx43 through the transfection of the Cx43 plasmid did not affect p-CaMKII and CaV3.2 expressions in vitro. Therefore, CaMKII and CaV3.2 may activate astrocytes by increasing [Ca2+]i, thereby mediating Cx43-dependent inflammation in VINP. Moreover, we demonstrated that the CaMKII signalling pathway was involved in VCR-induced inflammation, apoptosis, and mitochondrial damage. Collectively, our findings show a novel mechanism by which CaMKII and CaV3.2 mediate Cx43-dependent inflammation by activating astrocytes in neuropathic pain induced by VCR.
Graphical abstract
中文翻译：
CaMKII 和 CaV3.2 T 型钙通道通过激活长春新碱诱导的神经性疼痛中的星形胶质细胞介导 Connexin-43 依赖性炎症
长春新碱（VCR）是一种从长春花中分离出来的生物碱，是一种常用的化疗药物。然而，VCR疗法可导致剂量依赖性周围神经毒性，主要表现为神经性疼痛，这是限制其应用的主要原因之一。通过实验，我们发现VCR诱导的神经性疼痛（VINP）伴随着星形胶质细胞的激活；磷酸化 CaMKII (p-CaMKII)、Ca V 3.2 和 Connexin-43 (Cx43) 表达的上调；以及脊髓中炎性细胞因子和趋化因子的产生和释放。在星形胶质细胞培养中也观察到类似的情况。有趣的是，这些改变都可以通过鞘内注射 KN-93（一种 CaMKII 抑制剂）或l-抗坏血酸（一种 Ca V 3.2 抑制剂）来逆转。此外，KN-93和l-抗坏血酸抑制与星形胶质细胞活化相关的[Ca 2+ ] i增加。我们还证实，通过鞘内注射Cx43 siRNA或Gap27（一种Cx43模拟肽）敲低或抑制Cx43水平可缓解疼痛超敏反应并减少炎症因子的释放；然而，它们并不影响星形胶质细胞的激活或p-CaMKII 和Ca V 3.2 的表达。此外，通过Cx43质粒转染实现Cx43的过表达并不影响p-CaMKII和Ca V 3.2的体外表达。因此，CaMKII和Ca V 3.2可能通过增加[Ca 2+ ] i激活星形胶质细胞，从而介导VINP中Cx43依赖性炎症。此外，我们证明 CaMKII 信号通路参与 VCR 诱导的炎症、细胞凋亡和线粒体损伤。总的来说，我们的研究结果表明了一种新机制，即 CaMKII 和 Ca V 3.2 在 VCR 诱导的神经性疼痛中通过激活星形胶质细胞来介导 Cx43 依赖性炎症。
京公网安备 11010802027423号