Neuroprotective effects of Dendrobium alkaloids on rat cortical neurons injured by oxygen-glucose deprivation and reperfusion.
In this study we investigated the protective effects of alkaloids from Dendrobium spez. on cortical neurons injured by oxygen-glucose deprivation/reperfusion (OGD/RP) in vitro. Rat primary cultured cerebral cortical neurons were investigated at different time points of OGD/RP. The MTT assay and the lactate dehydrogenase (LDH) release were used to determine cell viability. The concentration of intracellular free calcium [Ca(2+)](i) and mitochondrial membrane potential (MMP) were determined to evaluate the degree of neuron damage. Morphologic changes of neurons following OGD/RP were examined by electron microscope. To evaluate neuron apoptosis, flow cytometry was performed and the expressions of caspase-3 and caspase-12 mRNA were examined by real-time quantitative PCR during OGD 2h/RP 12h. Treatment with Dendrobium alkaloids (0.025 approximately 2.5mg/l) significantly attenuated neuronal damage, with evidence of increased cell viability, decreased cell apoptosis, and decreased cell morphologic impairment. Furthermore, Dendrobium alkaloids inhibited [Ca(2+)](i) elevation, increased MMP and decreased the expressions of caspase-3 and caspase-12 in a concentration-dependent manner at OGD 2h/RP 12h. Dendrobium alkaloids have significantly protective effects on OGD/RP-induced neuronal damages in rat primary neuron cultures. The protection against OGD/RP-induced apoptosis appears to be mediated through blocking the decrease in MMP and increase in [Ca(2+)](i), as well as by down-regulating mRNA expression of caspase-3 and caspase-12.
Wang
Q
Q
Department of Pharmacology, Zunyi Medical College, 201 Dalian Road, Zunyi 563000, China. wangqianlily2006@gmail.com
Gong
Q
Q
Wu
Q
Q
Shi
J
J
D000470:Alkaloids / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000037:antagonists & inhibitors; D053454:Caspase 12 / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D031667:Dendrobium / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005947:Glucose; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology / Q000627:therapeutic use*; D010100:Oxygen; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology
D000470:Alkaloids; D018696:Neuroprotective Agents; D010936:Plant Extracts; D012333:RNA, Messenger; D007770:L-Lactate Dehydrogenase; D053454:Caspase 12; D053148:Caspase 3; D005947:Glucose; D010100:Oxygen; D002118:Calcium