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Published on: May 3, 2018
Levobupivacaine-induced vasoconstriction involves caldesmon phosphorylation mediated by tyrosine kinase-induced ERK
Soo Hee Lee1, Seong-Chun Kwon2, Seong-Ho Ok3
1Department of Anesthesiology and Pain Medicine, Gyeongsang National University College of Medicine, Gyeongsang National University Hospital, 15 Jinju-daero 816 beon-gil, Jinju-si, Gyeongsangnam-do 52727, Republic of Korea.
Abstract:
The goals of this study were to examine the cellular signaling pathways associated with the phosphorylation of caldesmon, the phosphorylation-dependent inhibitory protein of myosin phosphatase (CPI-17), and the 20-kDa regulatory light chain of myosin (MLC20) induced by levobupivacaine in isolated rat aortas. The effects of genistein, tyrphostin 23, GF109203X, PD98059, Y-27632, 1-butanol, and ML-7 HCl on levobupivacaine-induced contraction were assessed. The effect of genistein on the simultaneous calcium-tension curves induced by levobupivacaine was examined. The effects of GF109203X, genistein, PD98059 and extracellular signal-regulated kinase (ERK) siRNA on levobupivacaine-induced caldesmon phosphorylation were investigated. The effect of genistein on the ERK and tyrosine phosphorylation induced by levobupivacaine was examined. The effect of GF109203X, PD98059, Y-27632, SP600125, and ML-7 HCl on the levobupivacaine-induced phosphorylation of CPI-17 and MLC20 were investigated. Genistein, tyrphostin 23, GF109203X, PD98059, Y-27632, ML-7 HCl, and 1-butanol attenuated levobupivacaine-induced contraction. Genistein caused a right downward shift of the calcium-tension curves induced by levobupivacaine. Genistein attenuated levobupivacaine-induced phosphorylation of protein tyrosine, ERK and caldesmon. PD98059, ERK siRNA and GF109203X attenuated levobupivacaine-induced caldesmon phosphorylation. GF109203X, Y-27632, SP600125, ML-7 HCl and PD98059 attenuated CPI-17 phosphorylation and MLC20 phosphorylation induced by levobupivacaine. These results suggest that levobupivacaine-induced caldesmon phosphorylation contributing to levobupivacaine-induced contraction is mediated by a pathway involving ERK, which is activated by tyrosine kinase or protein kinase C (PKC). The phosphorylation of CPI-17 and MLC20 induced by levobupivacaine is mediated by cellular signaling pathways involving PKC, Rho-kinase, and c-Jun NH2-terminal kinase or PKC, Rho-kinase, ERK, and myosin light chain kinase.
Insights
Levobupivacaine causes aortic contraction via caldesmon, CPI-17, and MLC20 phosphorylation. Inhibitors like genistein and PD98059 block these pathways, suggesting therapeutic potential for levobupivacaine-induced vasoconstriction.
Area of Science:
- Pharmacology
- Cellular Biology
- Physiology
Background:
- Levobupivacaine, a local anesthetic, can induce vasoconstriction.
- Understanding the cellular mechanisms of levobupivacaine-induced contraction is crucial for managing its side effects.
Purpose of the Study:
- To investigate the cellular signaling pathways involved in levobupivacaine-induced phosphorylation of caldesmon, CPI-17, and MLC20 in rat aortas.
- To assess the effects of various inhibitors on levobupivacaine-induced contraction and related signaling events.
Main Methods:
- Isolated rat aortas were used to study levobupivacaine-induced contraction.
- Western blotting and siRNA were employed to investigate protein phosphorylation and signaling pathways.
- The effects of inhibitors targeting tyrosine kinase, PKC, ERK, Rho-kinase, and MLCK were examined.
Main Results:
- Levobupivacaine induced contraction and phosphorylation of caldesmon, CPI-17, and MLC20.
- Genistein, PD98059, GF109203X, Y-27632, ML-7 HCl, and 1-butanol attenuated levobupivacaine-induced contraction.
- Genistein inhibited tyrosine phosphorylation, ERK phosphorylation, and caldesmon phosphorylation.
- PD98059, ERK siRNA, and GF109203X inhibited caldesmon phosphorylation.
- GF109203X, PD98059, Y-27632, SP600125, and ML-7 HCl inhibited CPI-17 and MLC20 phosphorylation.
Conclusions:
- Levobupivacaine-induced contraction is mediated by caldesmon phosphorylation via ERK, activated by tyrosine kinase or PKC.
- CPI-17 and MLC20 phosphorylation involve PKC, Rho-kinase, and JNK/ERK/MLCK pathways.
- These findings elucidate the signaling mechanisms of levobupivacaine-induced vasoconstriction.
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