α2-adrenergic receptors in spiral ganglion neurons may mediate protective effects of brimonidine and yohimbine against glutamate and hydrogen peroxide toxicity.
Brimonidine, an alpha2-adrenergic receptor (α(2)-AR) agonist, is thought to be neuroprotective in some types of neurons via the activation of α(2)-AR. However, it is still unknown whether the α(2)-ARs exist in cochlear spiral ganglion neurons (SGNs). The authors aimed to demonstrate the presence and localization of α(2)-ARs in rat-cultured SGNs and to investigate the effect of brimonidine on glutamate- and hydrogen peroxide (H(2)O(2))-induced damage in the primary-cultured rat SGNs. The expression of α(2)-ARs was determined by reverse transcription-polymerase chain reaction, Western blot analysis and immunofluorescence. Then SGNs were exposed to glutamate or H(2)O(2) respectively with or without brimonidine. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Apoptosis was determined by acridine orange and Hoechst 33342/propidium iodide double staining. The protein expressions of α(2)-ARs, Bax, Bcl-2, Caspase-9, Caspase-3, p-ERK1/2, iNOS, and artemin were determined by Western blot respectively. The cell viability was markedly reduced after exposure of glutamate (1mM) or H(2)O(2) (300 μM) to SGNs. Treatment with brimonidine protected SGNs against glutamate- or H(2)O(2)-induced cell damage, enhanced SGNs survival, decreased the elevation of Bax, Caspase-9, Caspase-3, p-ERK1/2, and artemin triggered by glutamate or H(2)O(2), and altered the expressions of Bcl-2 and iNOS. These protective effects of brimonidine can be reversed by yohimbine. Overall, the study describes the localization of α(2)-ARs in rat-cultured SGNs and indicates that brimonidine, which may work directly via interaction with α(2)-ARs, attenuates glutamate- and H(2)O(2)-induced damage in SGNs by Caspase-dependent modes as well as Caspase-independent modes.
Otolaryngology-Head and Neck Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, PR China.
J
J
Cai
J
J
Li
W
W
Liu
Y
Y
Han
H
H
Wang
D000818:Animals; D000068438:Brimonidine Tartrate; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D018698:Glutamic Acid / Q000633:toxicity*; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000633:toxicity*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011810:Quinoxalines / Q000494:pharmacology*; D051381:Rats; D018341:Receptors, Adrenergic, alpha-2 / Q000502:physiology*; D013136:Spiral Ganglion / Q000187:drug effects / Q000502:physiology*; D015016:Yohimbine / Q000494:pharmacology*
D018696:Neuroprotective Agents; D011810:Quinoxalines; D018341:Receptors, Adrenergic, alpha-2; D015016:Yohimbine; D018698:Glutamic Acid; D000068438:Brimonidine Tartrate; D006861:Hydrogen Peroxide