title: Stimulation of beta2-adrenoceptors inhibits apoptosis in rat brain after transient forebrain ischemia.
abstract: We have previously demonstrated that the neuroprotective effect of the beta2-adrenoceptor agonist clenbuterol in vitro and in vivo was most likely mediated by an increased nerve growth factor (NGF) expression. In the present study, we examined whether clenbuterol was capable of inhibiting apoptosis caused by ischemia. Transient forebrain ischemia was performed in male Wistar rats (300 to 350 g) by clamping both common carotid arteries and reducing the blood pressure to 40 mm Hg for 10 minutes. Clenbuterol (0.1, 0.5, and 1.0 mg/kg intraperitoneally) was administered 3 hours before ischemia or immediately after ischemia. The brains were removed for histologic evaluation 7 days after ischemia. The time course of DNA fragmentation was determined 1, 2, 3 and 4 days after ischemia. Staining with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) was used for further analysis of DNA fragments in situ 3 days after ischemia. The NGF protein was assayed by enzyme-linked immunosorbent assay. Ten-minute forebrain ischemia damaged 80% to 90% of the neurons in the hippocampal CA1 region evaluated 7 days after ischemia. Pretreatment with clenbuterol (0.5 and 1.0 mg/kg) reduced the neuronal damage by 18.1% (P < 0.01) and 13.1% (P < 0.05), respectively. The neuroprotective effect also was found when clenbuterol (0.5 mg/kg) was administered immediately after ischemia (P < 0.05). The DNA laddering appeared in striatum 1 day and in hippocampus 2 days after ischemia and peaked on the third day in both regions. The DNA laddering was nearly abolished in the hippocampus and partially blocked in striatum and cortex by 0.5 mg/kg clenbuterol. These results were confirmed by TUNEL staining. Clenbuterol (0.5 mg/kg intraperitoneally) elevated the NGF protein level by 33% (P < 0.05) in the hippocampus and 41% (P < 0.05) in the cortex 6 hours after ischemia. Three days after ischemia, the NGF levels in these regions were no longer different between the clenbuterol-treated and control groups. This study clearly demonstrates that clenbuterol possesses a neuroprotective activity and a marked capacity to inhibit DNA degradation after global ischemia. The results suggest that clenbuterol increases NGF expression during the first hours after global ischemia and thereby protects neurons against apoptotic damage.
authors.affiliation: Institut für Pharmakologie und Toxikologie, Fachbereich Pharmazie und Lebensmittelchemie, Philipps-Universität, Marburg, Germany.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhu
authors.forename: C
authors.initials: C
authors.lastname: Culmsee
authors.forename: I
authors.initials: I
authors.lastname: Semkova
authors.forename: J
authors.initials: J
authors.lastname: Krieglstein
mesh_terms: D000318:Adrenergic beta-Agonists / Q000627:therapeutic use*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001921:Brain / Q000187:drug effects* / Q000473:pathology; D002976:Clenbuterol / Q000627:therapeutic use*; D053938:DNA Fragmentation; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D002546:Ischemic Attack, Transient / Q000188:drug therapy*; D008297:Male; D009414:Nerve Growth Factors / Q000096:biosynthesis; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D016548:Prosencephalon / Q000098:blood supply*; D051381:Rats; D017208:Rats, Wistar; D013268:Stimulation, Chemical
chemical_list: D000318:Adrenergic beta-Agonists; D003856:Deoxyuracil Nucleotides; D009414:Nerve Growth Factors; D004253:DNA Nucleotidylexotransferase; D002976:Clenbuterol