Is activation of AMP-activated protein kinase involved in ropivacaine-induced SH-SY5Y cell apoptosis? / 中华麻醉学杂志

ABSTRACT

Objective To determine if activation of AMP-activated protein kinase (AMPK) is involved in ropivacaine-induced reactive oxygen species (ROS) production and apoptosis in human neuroblastoma cell line SHSY5Y.Methods SH-SY5Y cell line was purchased from cell center of Shanghai life Science Research Institute,Chinese Academy of Sciences and cultured in DMEM/F12 liquid culture medium containing 15 % bovine calf serum at 37 ℃ in incubator filled with 5% CO2 . Plasmids pGPU6/GFP/Neo-shRNA AMPKα2 and pEGFP-N1-AMPKα2were transfected into the SH-SY5Y cell line. The expression of AMPKα2 was determined by Western blot analysis.The SH-SY5Y cells transfected with recombinant plasmids were exposed to 3 mol/L ropivacaine. Intracellular ROS was detected by flow cytometry. Cell viability was quantitatively determined by MTT colorimetry assay. Apoptosis was assessed by flow cytometry and Hoechst33258 staining. Results The plasmid pEGFP-N1-AMPKα2 upregulated while pGPU6/GFP/Neo-shRNA AMPKα2 down-regulated the expression of AMPKα2 ( P < 0.01). Down-regulation of AMPKα2 expression attenuated while up-regulation of. AMPKα2 expression promoted intracellular ROS production and cell apoptosis induced by ropivacaine ( P < 0.01) . Conclusion AMPK probably mediates ROS production and cell apoptosis induced by ropivacaine.