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Aim: To investigate whether a similar process mediates cytotoxicity of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3, edelfosine) in both yeasts and human tumor cells., Methods: A modified version of a previously described assay for the intracellular conversion of nitro blue tetrazolium to formazan by superoxide anion was used to measure the generation of reactive oxygen species (ROS). Apoptotic yeast cells were detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. DNA fragmentation and the generation of ROS were measured by cytofluorimetric analysis in Jurkat cells., Results: Edelfosine induced apoptosis in Saccharomyces cerevisiae, as assessed by TUNEL assay. Meanwhile, edelfosine induced a time- and concentration-dependent generation of ROS in yeasts. Rotenone, an inhibitor of the mitochondrial electron transport chain, prevented ROS generation and apoptosis in response to edelfosine in S cerevisiae. alpha-Tocopherol abrogated the edelfosine-induced generation of intracellular ROS and apoptosis. Edelfosine also induced an increase of ROS in human leukemic cells that preceded apoptosis. The overexpression of Bcl-2 by gene transfer abrogated both ROS generation and apoptosis induced by edelfosine in leukemic cells. Changes in the relative mitochondrial membrane potential were detected in both yeasts and Jurkat cells., Conclusion: These results indicate that edelfosine induces apoptosis in yeasts in addition to human tumor cells, and this apoptotic process involves mitochondria, likely through mitochondrial-derived ROS. These data also suggest that yeasts can be used as a suitable cell model in elucidating the antitumor mechanism of action of edelfosine.

Aim: To study that l-stepholidine (SPD) regulates the expression of proenkephalin (PENK) and prodynorphin (PDYN) mRNA and Fos in the striatum after rotational test in the 6-hydroxydopamine (6-OHDA)-lesioned rats., Methods: PENK and PDYN mRNA levels were examined with in situ hybridization, and Fos expression was detected with immunocytochemistry. The data were semi-quantified with image analyzer., Results: (1) Following repeated SPD treatment, the rotation was kept on high activity in the 6-OHDA-lesioned rats. (2) SPD significantly elicited Fos expression in both sides of striatum, particularly in the denervated one. Repeated administration of SPD, Fos expression declined on both sides, particularly in the intact one. (3) In the denervated striatum of 6-OHDA-lesioned rats, the PENK mRNA level was extremely increased vs that in the intact striatum. This high level of PENK mRNA was significantly reduced by 7-d treatments of SPD. SPD also reduced the level of PENK mRNA in the intact striatum. However, the level of PDYN mRNA did not show significant change in both sides of striatum after denervation or SPD treatment., Conclusion: In the 6-OHDA-lesioned rats, the rotation induced by SPD was kept on a high activity, which was in pace with the inducement of Fos expression and the reduction of expression of PENK mRNA in the denervated striatum. But then the lesion and SPD treatment had no remarkable effect on the expression of PDYN mRNA.

Aim: To assess the atypical neuroleptic properties of a novel antipsychotic agent, olanzapine (Ola)., Methods: The action of Ola on apomorpine (Apo)-induced climbing behavior, 5-hydroxy-dl-tryptophan (5-HTP)-induced head twitch response, oxotremorine-induced tremor, and the conditioned avoidance behavior in mice were observed. The catalepsy of mice induced by Ola was also investigated. The single unit extracellular recording technique was used to compare the spontaneous firing rate changes of dopamine (DA) cells in the ventral tegmental area (VTA, A10) and the substantia nigra pars compact (SNC, A9) in rats after i.v. Ola., Results: Ola antagonized the climbing behavior (ED50 1.8 mg.kg-1, p.o.), head twitch behavior (ED50 0.3 mg.kg-1, p.o.), and tremor (ED50 5.2 mg.kg-1, p.o.) in mice. In a conditioned avoidance paradigm in mice, Ola inhibited the avoidance response with an ED50 of 2.72 mg.kg-1 (p.o.). However, the catalepsy was not induced by Ola in mice even under a very high dose of 100 mg.kg-1 (p.o.). Ola selectively increased the firing rate of DA cells in the VTA, but failed to affect that of SNC DA cells., Conclusion: Ola distinguished itself from the typical neuroleptic (e.g. haloperidol, Hal) and took resemblance of the atypical neuroleptic (e.g. clozapine, Clo) in 3 aspects: 1) the multiple receptor pharmacodynamics involving D1/D2, 5-HT2 and M-ACh receptors; 2) dose-response separation between the block of conditioned avoidance response and catalepsy induction; and 3) the specificity of action sites of firing rates upon acute drug challenge.