Department of Anesthesiology, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2022 Apr 28;47(4):431-442. doi: 10.11817/j.issn.1672-7347.2022.200991.
There are clinical reports of nerve injury caused by ropivacaine. The mechanism for nerve injury induced by ropivacaine has not been fully clarified. This study aims to investigate the changes of pain threshold and L spinal cord genomics at 6 h and 24 h after intrathecal injection of 0.5% and 1.0% ropivacaine, and to explore the underlying mechanisms for nerve injury caused by ropivacaine.
A total of 30 male Sprague Dawley rats weighing 220-260 g were successfully implanted with microspinal catheter. The rats were randomly divided into 5 groups (each =6): a control group (given saline), a ropivacaine group 1 and a ropivacaine group 2 (both given 1% ropivacaine), a ropivacaine group 3 and a ropivacaine group 4 (both given 0.5% ropivacaine). The rats received continuous intrathecal injection of corresponding drugs at 8.3 μL/h for 24 h via an implanted intrathecal catheter followed by 24 h-pause of injection for the ropivacaine group 2, the ropivacaine group 4 and the control group, 6 h-pause of injection for the ropivacaine group 1 and the ropivacaine group 3. For each group, the observation of behavioral change and the paw withdrawal mechanical threshold (PWMT) was conducted immediately after the injection and again after the pause of injection. After the PWMT observation, the rats were dissected to acquire L spinal cords. Illumina sequencing was applied to construct gene libraries. Then the statistical methods were used to find out differentially expressed genes between the groups. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis were conducted for those genes. Real-time RT-PCR was used to determine different expressions of some of those genes.
Compared with control group, the PWMT got higher in the ropivacaine group 1-4 and was positively correlated with concentration, negatively correlated with discontinuation duration. Compared with control group, the ropivacaine group 1 had 488 differentially expressed genes, of which 456 were up-regulated and 32 were down-regulated; the ropivacaine group 2 had 1 194 differentially expressed genes, of which 1 092 were up-regulated and 102 were down-regulated; the ropivacaine group 3 had 518 differentially expressed genes, of which 384 were up-regulated and 134 were down-regulated; and the ropivacaine group 4 had 68 differentially expressed genes, of which 46 were up-regulated and 22 were down-regulated. GO enrichment analysis and KEGG signaling pathway analysis showed that most of these differentially expressed genes were related to signaling pathways of inflammatory response.
After intrathecal injection of 0.5% ropivacaine and 1.0% ropivacaine for 24 h, the differentially expressed genes in L spinal cord of rats are mainly related to signaling pathways of inflammatory response.
有临床报告称罗哌卡因会导致神经损伤。罗哌卡因引起神经损伤的机制尚未完全阐明。本研究旨在探讨鞘内注射 0.5%和 1.0%罗哌卡因后 6 h 和 24 h 时疼痛阈值和 L 脊髓基因组的变化，并探讨罗哌卡因引起神经损伤的潜在机制。
30 只雄性 Sprague Dawley 大鼠（体重 220-260 g）成功植入微脊髓导管。大鼠随机分为 5 组（每组=6）：对照组（给予生理盐水）、罗哌卡因组 1 和罗哌卡因组 2（均给予 1%罗哌卡因）、罗哌卡因组 3 和罗哌卡因组 4（均给予 0.5%罗哌卡因）。通过植入的鞘内导管以 8.3 μL/h 的速度连续鞘内注射相应药物 24 h，然后对罗哌卡因组 2、罗哌卡因组 4 和对照组暂停注射 24 h，对罗哌卡因组 1 和罗哌卡因组 3 暂停注射 6 h。对于每组，在注射后立即和再次暂停注射后观察行为变化和足底机械缩足阈值（PWMT）。观察 PWMT 后，解剖大鼠获取 L 脊髓。应用 Illumina 测序构建基因文库。然后使用统计学方法找出组间差异表达的基因。对这些基因进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）信号通路分析。实时 RT-PCR 用于确定其中一些基因的不同表达。
与对照组相比，罗哌卡因组 1-4 的 PWMT 升高，且与浓度呈正相关，与停药时间呈负相关。与对照组相比，罗哌卡因组 1 有 488 个差异表达基因，其中 456 个上调，32 个下调；罗哌卡因组 2 有 1194 个差异表达基因，其中 1092 个上调，102 个下调；罗哌卡因组 3 有 518 个差异表达基因，其中 384 个上调，134 个下调；罗哌卡因组 4 有 68 个差异表达基因，其中 46 个上调，22 个下调。GO 富集分析和 KEGG 信号通路分析表明，这些差异表达基因主要与炎症反应信号通路有关。
鞘内注射 0.5%和 1.0%罗哌卡因 24 h 后，大鼠 L 脊髓中差异表达的基因主要与炎症反应信号通路有关。
