Journal of Pain Research (May 2023)
Pulsed Radiofrequency Decreases pERK and Affects Intracellular Ca2+ Influx, Cytosolic ATP Level, and Mitochondrial Membrane Potential in the Sensitized Dorsal Root Ganglion Neuron Induced by N-Methyl D-Aspartate
Abstract
Ristiawan Muji Laksono,1,2 Handono Kalim,3 Mohammad Saifur Rohman,4 Nashi Widodo,5 Muhammad Ramli Ahmad,6 Willy Halim7 1Doctoral Program in Biomedical Science, Faculty of Medicine, Brawijaya University, Malang, Indonesia; 2Department of Anesthesiology and Intensive Therapy, Faculty of Medicine, Brawijaya University, Malang, Indonesia; 3Department of Internal Medicine, Faculty of Medicine, Brawijaya University, Malang, Indonesia; 4Department of Cardiology and Vascular Medicine, Faculty of Medicine, Brawijaya University, Malang, Indonesia; 5Department of Biology, Faculty of Mathematics and Natural Science, Brawijaya University, Malang, Indonesia; 6Department of Anesthesiology, Intensive Care and Pain Management, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia; 7Faculty of Medicine, Brawijaya University, Malang, IndonesiaCorrespondence: Ristiawan Muji Laksono, Department of Anesthesiology and Intensive Therapy, Faculty of Medicine, Brawijaya University, Jl. Jaksa Agung Suprapto No. 2, Malang, East Java, Indonesia, Tel +62 812-3377-3593, Email [email protected]: The molecular mechanism of pulsed radiofrequency (PRF) in chronic pain management is not fully understood. Chronic pain involves the activation of specific N-Methyl D-Aspartate receptors (NMDAR) to induce central sensitization. This study aims to determine the effect of PRF on central sensitization biomarker phosphorylated extracellular signal-regulated kinase (pERK), Ca2+ influx, cytosolic ATP level, and mitochondrial membrane potential (Δψm) of the sensitized dorsal root ganglion (DRG) neuron following NMDAR activation.Methods: This study is a true experimental in-vitro study on a sensitized DRG neuron induced with 80 μM NMDA. There are six treatment groups including control, NMDA 80 μM, Ketamine 100 μM, PRF 2Hz, NMDA 80 μM + PRF 2 Hz, and NMDA 80 μM + PRF 2 Hz + ketamine 100 μM. We use PRF 2 Hz 20 ms for 360 seconds. Statistical analysis was performed using the One-Way ANOVA and the Pearson correlation test with α=5%.Results: In the sensitized DRG neuron, there is a significant elevation of pERK. There is a strong correlation between Ca2+, cytosolic ATP level, and Δψm with pERK intensity (p< 0.05). PRF treatment decreases pERK intensity from 108.48 ± 16.95 AU to 38.57 ± 5.20 AU (p< 0.05). PRF exposure to sensitized neurons also exhibits a Ca2+ influx, but still lower than in the unexposed neuron. PRF exposure in sensitized neurons has a higher cytosolic ATP level (0.0458 ± 0.0010 mM) than in the unexposed sensitized neuron (0.0198 ± 0.0004 mM) (p< 0.05). PRF also decreases Δψm in the sensitized neuron from 109.24 ± 6.43 AU to 33.21 ± 1.769 AU (p< 0.05).Conclusion: PRF mechanisms related to DRG neuron sensitization are by decreasing pERK, altering Ca2+ influx, increasing cytosolic ATP level, and decreasing Δψm which is associated with neuron sensitization following NMDAR activation.Keywords: pulsed radiofrequency, neurons, sensitization, calcium, ATP, mitochondrial membrane potential, NMDA, NMDAR
