STAT3 has neuroprotective effect via non-canonical activation and mitochondrial translocation, but its effect on ropivacaine-induced neurotoxicity remains unclear. Our previous study revealed that apoptosis was an important mechanism of ropivacaine-induced neurotoxicity; this study is to illustrate the relationship between STAT3 with ropivacaine-induced apoptosis. Those results showed that ropivacaine treatment decreased cell viability, induced cell cycle arrest in the G0/G1 phase, apoptosis, oxidative stress, and mitochondrial dysfunction in PC12 cells. Moreover, ropivacaine decreased the phosphorylated levels of STAT3 at Ser727 and downregulated the expression of STAT3 upstream gene IL-6. The mitochondrial translocation of STAT3 was also hindered by ropivacaine. To further illustrate the connection of STAT3 protein structure with ropivacaine, the autodock-vina was used to examine the interaction between STAT3 and ropivacaine, and the results showed that ropivacaine could bind to STAT3’s proline site and other sites. In addition, the activator and inhibitor of mitoSTAT3 translocation were used to demonstrate it was involved in ropivacaine-induced apoptosis; the results showed that enhancing the mitochondrial STAT3 translocation could prevent ropivacaine-induced apoptosis. Finally, the expression of p-STAT3 and the levels of apoptosis in the spinal cord were also detected; the results were consistent with the cell experiment; ropivacaine decreased the expression of p-STAT3 protein and increased the levels of apoptosis in the spinal cord. We demonstrated that ropivacaine induced apoptosis by inhibiting the phosphorylation of STAT3 at Ser727 and the mitochondrial STAT3 translocation. This effect was reversed by the activation of the mitochondrial STAT3 translocation.

STAT3 通过非经典激活和线粒体易位具有神经保护作用，但其对罗哌卡因诱导的神经毒性的影响仍不清楚。我们之前的研究表明，细胞凋亡是罗哌卡因神经毒性的重要机制。本研究旨在阐明STAT3与罗哌卡因诱导的细胞凋亡的关系。这些结果表明，罗哌卡因治疗降低了 PC12 细胞的细胞活力、诱导细胞周期停滞在 G0/G1 期、细胞凋亡、氧化应激和线粒体功能障碍。此外，罗哌卡因降低了 STAT3 在 Ser727 位点的磷酸化水平，并下调了 STAT3 上游基因 IL-6 的表达。罗哌卡因也阻碍了 STAT3 的线粒体易位。为了进一步说明 STAT3 蛋白结构与罗哌卡因的联系，使用autodock-vina检测STAT3与罗哌卡因的相互作用，结果表明罗哌卡因可以与STAT3的脯氨酸位点及其他位点结合。此外，利用mitoSTAT3易位的激活剂和抑制剂证明其参与了罗哌卡因诱导的细胞凋亡；结果表明，增强线粒体STAT3易位可以防止罗哌卡因诱导的细胞凋亡。最后，还检测了脊髓中p-STAT3的表达和凋亡水平；结果与细胞实验一致；罗哌卡因降低了 p-STAT3 蛋白的表达，增加了脊髓细胞凋亡的水平。我们证明罗哌卡因通过抑制 STAT3 在 Ser727 位点的磷酸化和线粒体 STAT3 易位来诱导细胞凋亡。