Dehydrocostuslactone protects against oxygen-glucose deprivation/reperfusion-induced injury by inhibiting autophagy and apoptosis in PC12 cells

OBJECTIVE TO investigate the neural protection of dehydrocostus lactone(DHL)against neuronal injury induced by oxygen and glucose deprivation/reperfusion(OGD/R)in differentiated PC12 cells.METHODS We used a cellular model of 2 h of OGD and 24 h of reperfusion to mimic cerebral ischemia-reperfusion injury.Cell viability was used to reflect the degree of OGD/R-induced injury.Cells were treated with DHL during the reperfusion phase.Cell Counting Kit(CCK-8)and LDH assays were performed to determine the optimal dose of DHL and cell viability.Flow cytometry analysis and Monodansylcadaverine(MDC)staining were then conducted to detect apoptosis rate and autophagosome formation after OGD/R in PC12 cells.Immunofluorescence and Western blotting analyses were used to detect the expres⁃sion of proteins associated with autophagy and apoptosis.RESULTS OGD/R significantly decreased cell viability and increased apoptosis rate.The expression levels of autophagy-related proteins,namely,LC3 and Beclin-1,and apoptosisrelated proteins,namely,Bax and caspase-3 increased,but that of the anti-apoptosis Bcl-2 protein decreased.However,DHL attenuated OGD/R-induced neuronal injury through inhibition of apoptosis and autophagy properties by modulating au⁃tophagy-associated proteins(LC3 and Beclin-1)and apoptosis-modulating proteins(caspase-3 and Bcl-2/Bax).CONCLU⁃SION Our data provide an evidence for the neuroprotective effect of DHL against ischemic neuronal injury.Hence,DHL could be a promising candidate for treatment of ischemic stroke.

Rapid Determination of Costunolide and Dehydrocostuslactone in Human Plasma Sample and Chinese Patent Medicine Xiang Sha Yang Wei Capsule Using HPLC-DAD Coupled with Second-order Calibration

A novel methodology that combines high performance liquid chromatography with photodiode-array detector （HPLC-DAD） coupled with second-order calibration method based on alternating trilinear decomposition （ATLD） algorithm was used in determination of the effective constituents such as costunolide and dehydrocostuslactone, in plasma sample and Chinese patent medicine Xiang Sha Yang Wei （XSYW） capsule. Complicated systems such as plasma and Chinese patent medicine which have intricate components are tedious to isolate and purify. The problem that chromatographic peaks are heavily overlapped among the analytes and interferents from the background matrices can be resolved, and the satisfactory quantification results have been gained with the help of the ATLD algorithm which utilized ＂mathematical separation＂ instead of partial ＂physical or chemical separation＂. Meanwhile, HPLC-MS/MS method was used to validate the accuracy of the proposed determination method.