Bupivacaine reduces GlyT1 expression by potentiating the p-AMPKα/BDNF signalling pathway in spinal astrocytes of rats.

Abstract

Bupivacaine, a local anaesthetic, is widely applied in the epidural or subarachnoid space to clinically manage acute and chronic pain. However, the underlying mechanisms are complex and unclear. Glycine transporter 1 (GlyT1) in the spinal cord plays a critical role in various pathologic pain conditions. Therefore, we sought to determine whether bupivacaine exerts its analgesic effect by regulating GlyT1 expression and to determine the underlying mechanisms of regulation. Primary astrocytes prepared from the spinal cord of rats were treated with bupivacaine. The protein levels of GlyT1, brain-derived neurotrophic factor (BDNF) and phosphorylated adenosine 5'-monophosphate (AMP)-activated protein kinase α (p-AMPKα) were measured by western blotting or immunofluorescence. In addition, 7,8-dihydroxyflavone (7,8-DHF, BDNF receptor agonist) and AMPK shRNA were applied to verify the relationship between the regulation of GlyT1 by bupivacaine and the p-AMPKα/BDNF signalling pathway. After treatment with bupivacaine, GlyT1 expression was diminished in a concentration-dependent manner, while the expression of BDNF and p-AMPK was increased. Moreover, 7,8-DHF decreased GlyT1 expression, and AMPK knockdown suppressed the upregulation of BDNF expression by bupivacaine. Finally, we concluded that bupivacaine reduced GlyT1 expression in spinal astrocytes by activating the p-AMPKα/BDNF signalling pathway. These results provide a new mechanism for the analgesic effect of intrathecal bupivacaine in the treatment of acute and chronic pain.

• Authors: Kaimei Lu, Liyan Zhao, Yonghai Zhang, Fan Yang, Huiwen Zhang, Jie Wang, Bin Li, Guimei Ji, Jianqiang Yu, Hanxiang Ma
• Journal: Scientific reports (Sci Rep) Vol. 12 Issue 1 Pg. 1378 (01 26 2022) ISSN: 2045-2322 [Electronic] England
• PMID: 35082359 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2022. The Author(s).

Chemical References

• 6,7-dihydroxyflavone
• Anesthetics, Local
• Bdnf protein, rat
• Brain-Derived Neurotrophic Factor
• Flavones
• Glycine Plasma Membrane Transport Proteins
• Slc6a9 protein, rat
• Receptor, trkB
• AMP-Activated Protein Kinases
• Bupivacaine

Topics

• AMP-Activated Protein Kinases (genetics, metabolism)
• Anesthetics, Local (pharmacology)
• Animals
• Animals, Newborn
• Astrocytes (drug effects, metabolism)
• Brain-Derived Neurotrophic Factor (metabolism)
• Bupivacaine (pharmacology)
• Cells, Cultured
• Flavones (pharmacology)
• Gene Knockdown Techniques (methods)
• Glycine Plasma Membrane Transport Proteins (metabolism)
• Rats
• Rats, Sprague-Dawley
• Receptor, trkB (agonists)
• Signal Transduction (drug effects, genetics)
• Spinal Cord (cytology)
• Transfection