Dezocine protects primary rat cortical astrocyte against β amyloid peptide exposure: roles of hyperpolarization-activated cyclic nucleotide-gated channels / 临床麻醉学杂志

ABSTRACT

Objective To investigate hyperpolarization-activated cyclic nucleotide-gated (HCN) channels inhibited by dezocine when primary rat cortical astrocytes were exposed to β amyloid peptide.Methods Cultured primary cortical astrocytes from new-born SD rats (within 24 h) were divided into 7 groups (n=5) according to random number table.The astrocytes in groups ZA and CDA were pretreated with 7-CH-cAMP (30 mmol/L),an agonist of HCN channels and ZD7288 (30 mmol/L),an inhibitor of HCN channels for 24 h,respectively.And then the cells in groups DA1,DA2,DA3 and CDA were treated with dezocine 10-7,10-6,10-5and 10-16 mmol/L for 24 h,respectively.Following with the treatments above,the cells in groups A,DA1,DA2,DA3,ZA and CDA were exposed to Aβ1-42 (15μmol/L) for 24 h,but the cells in group C were cultured normally.The effects of cell apoptosis,viability and injury were assessed by Annexin V-FITC/PI assay,MTT assay and lactate de hydrogenase (LDH) release assay.IL-1β was assessed by ELISA assay.The expressions of Cu/ZnSOD and Mn-SOD protein were assessed by Western blot assay.Results Compared with group C,there were significant increases of cell apoptosis,injury and IL-1β level in group A (P＜0.05).Compared with group A,there were significant decreases of cell apoptosis,injury,and IL-1β level but increase of Cu/Zn-SOD and Mn-SOD expressions in groups ZA,DA2 and DA3 (P＜0.05),while the neuroprotection of dezocine was partially restored by 7-CH-cAMP in group CDA (P ＜ 0.05).Conclusion The neuroprotection of dezocine against apoptosis induced by β amyloid peptide could be associated with up-regulation of anti-oxidative stress related Cu/Zn-SOD and Mn-SOD mediated by inhibition of HCN channels.