Introduction

Ropivacaine has been regularly used because of its good anesthetic and analgesic effects, but it may exert neurotoxic effects on neurocyte. Dexmedetomidine has presented special advantages in the fields of neuroprotection, and it also could improve peripheral nerve block combining with ropivacaine. However, if dexmedetomidine could repair neurocyte injury induced by ropivacaine, and the specific mechanism remain unclear.

Methods

Western blotting and qRT-PCR were applied for measuring expression of protein and mRNA, respectively. Flow cytometry was used for assessing apoptosis. Cell proliferation was detected using Cell Counting Kit-8 (CCK-8) and colony formation assays. Transwell assay was applied to measure the migration and invasion of cells. Dual luciferase reporter assay was applied for confirming the binding site between microRNA-381 (miR-381) and Leucine-rich repeat C4 protein (LRRC4).

Results

The viability of PC12 cells increased with raising the concentration of dexmedetomidine (0 μM, 10 μM, 50 μM, 100 μM). Dexmedetomidine reversed role of ropivacaine (0 mM, 0.1 mM, 0.5 mM, 1 mM) by upragulating the expression of miR-381 and suppressing the expression of LRRC4 in PC12 cells. miR-381 can directly interact with target gene LRRC4 and negatively regulate its expression. Dexmedetomidine promoted the proliferation, migration, and invasion and inhibited apoptosis of PC12 cells by suppressing LRRC4 via up-regulating the expressions of miR-381 and further activated SDF-1/CXCR4 signaling pathway.

Conclusions

Dexmedetomidine could protect PC12 cells from ropivacaine injury through miR-381/LRRC4/SDF-1/CXCR4 signaling pathway. This study may provide new therapeutic strategy targeting miR-381/LRRC4/SDF-1/CXCR4 signaling pathway about the prevention of ropivacaine induced neurocyte injury.

中文翻译：

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右美托咪定通过 miR-381/LRRC4 /SDF-1/CXCR4 信号通路保护 PC12 细胞免受罗哌卡因损伤。

介绍

罗哌卡因因其良好的麻醉和镇痛作用而被经常使用，但它可能对神经细胞产生神经毒性作用。右美托咪定在神经保护领域具有特殊优势，与罗哌卡因联合使用可改善周围神经阻滞。但右美托咪定能否修复罗哌卡因所致的神经细胞损伤，具体机制尚不清楚。

方法

Western印迹和qRT-PCR分别用于测量蛋白质和mRNA的表达。流式细胞术用于评估细胞凋亡。使用 Cell Counting Kit-8 (CCK-8) 和集落形成测定法检测细胞增殖。Transwell 测定用于测量细胞的迁移和侵袭。应用双荧光素酶报告基因测定来确认 microRNA-381 (miR-381) 和富含亮氨酸的重复 C4 蛋白 (LRRC4) 之间的结合位点。

结果

PC12细胞的活力随着右美托咪定（0 μM、10 μM、50 μM、100 μM）浓度的增加而增加。右美托咪定通过上调 miR-381 的表达和抑制 LRRC4 在 PC12 细胞中的表达来逆转罗哌卡因（0 mM、0.1 mM、0.5 mM、1 mM）的作用。miR-381可以直接与靶基因LRRC4相互作用并负调控其表达。右美托咪定通过上调miR-381的表达抑制LRRC4，进一步激活SDF-1/CXCR4信号通路，促进PC12细胞的增殖、迁移和侵袭，抑制细胞凋亡。

结论

右美托咪定可通过miR-381/LRRC4/SDF-1/CXCR4信号通路保护PC12细胞免受罗哌卡因损伤。本研究可能为预防罗哌卡因诱导的神经细胞损伤提供靶向miR-381/LRRC4/SDF-1/CXCR4信号通路的新治疗策略。