Department of Anesthesiology, Beijing Shijitan Hospital, Capital Medical University, 10 Tieyi Rd, Haidian District, Beijing, 100038, China.
Neurotox Res. 2020 Jun;38(1):175-183. doi: 10.1007/s12640-020-00202-3. Epub 2020 Apr 25.
Clinical application of local anesthetic reagent, liposomal bupivacaine (BUP), may cause irreversible damage to human nerve system. In this study, we explored the functional role of long non-coding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) in BUP-induced neurotoxicity in SH-SY5Y cells. SH-SY5Y cells were treated with BUP in vitro, whose dose-dependent effects on cell viability and SNHG16 expression were explored. SNHG16 was upregulated in SH-SY5Y cells. The protection of SNHG16 upregulation on BUP-induced neurotoxicity was examined by viability assay, apoptosis assay, and caspase activity assay, respectively. The endogenously competing target of SNHG16, human mature microRNA-132-3p (hsa-miR-132-3p), was explored by dual-luciferase assay and quantitative real-time PCR (qRT-PCR). Hsa-miR-132-3p was then further overexpressed in SNHG16-upregulated SH-SY5Y cells to explore its functional role in BUP-induced neurotoxicity. BUP induced dose-dependent cell death and SNHG16 downregulation in SH-SY5Y cells. Inversely, lentivirus-mediated SNHG16 upregulation mitigated cell death. In addition, SNHG16 upregulation rescued BUP-induced apoptosis and caspase 3/7 augmentation. Hsa-miR-132-3p was found to be reversely expressed with SNHG16 in BUP-treated SH-SY5Y cells. Overexpressing hsa-miR-132-3p reduced the protection of SNHG16 on BUP-induced neurotoxicity. We demonstrated that epigenetic axis of SNHG16/hsa-miR-132-3p had a functional role in regulating anesthesia-induced neurotoxicity in human lineage neural cells.
局部麻醉剂脂质体布比卡因（BUP）的临床应用可能对人体神经系统造成不可逆转的损害。在这项研究中，我们探讨了长链非编码 RNA（lncRNA）小核仁 RNA 宿主基因 16（SNHG16）在 BUP 诱导的 SH-SY5Y 细胞神经毒性中的功能作用。我们在体外用 BUP 处理 SH-SY5Y 细胞，探讨了其对细胞活力和 SNHG16 表达的剂量依赖性影响。结果显示 SNHG16 在 SH-SY5Y 细胞中上调。通过细胞活力测定、细胞凋亡测定和 caspase 活性测定分别检测 SNHG16 上调对 BUP 诱导的神经毒性的保护作用。通过双荧光素酶报告基因实验和定量实时 PCR（qRT-PCR）检测 SNHG16 的内源性竞争靶标，人类成熟 microRNA-132-3p（hsa-miR-132-3p）。然后在 SNHG16 上调的 SH-SY5Y 细胞中进一步过表达 hsa-miR-132-3p，以探讨其在 BUP 诱导的神经毒性中的功能作用。BUP 诱导 SH-SY5Y 细胞呈剂量依赖性死亡和 SNHG16 下调。相反，慢病毒介导的 SNHG16 上调减轻了细胞死亡。此外，SNHG16 上调挽救了 BUP 诱导的细胞凋亡和 caspase 3/7 增加。在 BUP 处理的 SH-SY5Y 细胞中发现 hsa-miR-132-3p 与 SNHG16 呈反向表达。过表达 hsa-miR-132-3p 降低了 SNHG16 对 BUP 诱导的神经毒性的保护作用。我们证明了 SNHG16/hsa-miR-132-3p 的表观遗传轴在调节人源性神经细胞麻醉诱导的神经毒性中具有功能作用。
