Yingliang Wu

Effect of MK-801 and ketamine on hydroxyl radical generation in the posterior cingulate and retrosplenial cortex of free-moving mice, as determined by in vivo microdialysis

This study investigated the effect of MK-801 and ketamine, N-methyl-D-aspartate (NMDA) receptor antagonists which can induce schizophrenic symptoms and have neurotoxicity in human and animals, on hydroxyl radical (*OH) generation in the posterior cingulate and retrosplenial (PC/RS) cortex of free-moving mice using the salicylic acid trapping technique. MK-801 (0.6 mg/kg) or ketamine (50 mg/kg) acute administration significantly increased *OH levels in mouse PC/RS cortex. The basal *OH levels after MK-801 and ketamine administrations for 7 consecutive days were significantly increased compared with the naive basal levels. MK-801 (0.6 mg/kg) or ketamine (50 mg/kg) challenge after chronic administration further significantly increased dialysate levels of *OH. Our study also found that the release of *OH was secondary to stereotyped behavior, and the intensity of stereotyped behavior induced by MK-801 was more than that induced by ketamine. The results suggested that NMDA receptor antagonists participate in the generation of *OH in the PC/RS cortex of mouse, and oxidative stress, derived from the formation of free radicals, might play an important role in the pathophysiology of these two models of schizophrenia.

Psychological stress selectively increases extracellular dopamine in the 'shell', but not in the 'core' of the rat nucleus accumbens : a novel dual-needle probe simultaneous microdialysis study

In order to compare psychological stress-induced dopamine (DA) release in two subterritories (e.g. shell and core) of the nucleus accumbens of the same animal, a novel dual-needle microdialysis probe has been developed. The two needles were placed in the ipsilateral shell and core subterritories of the nucleus accumbens under pentobarbital anesthesia and 24 h later the microdialysis was started. Basal DA output was not significantly different between the shell and the core. Psychological stress for 20 min significantly increased extracellular DA levels in the shell of the nucleus accumbens, however, the levels of dopamine remained almost unaltered in the core. This finding suggests that DA transmission in the shell of the nucleus accumbens was selectively activated during psychological stress, and that the shell plays an important role in emotional responses. The results further show that microdialysis using the novel dual-needle probe could be very useful to differentiate neurochemical changes occurring in neighboring areas in the brain.

Synthesis and evaluation of benzimidazole carbamates bearing indole moieties for antiproliferative and antitubulin activities

A series of novel benzimidazole carbamates bearing indole moieties with sulphur or selenium atoms connecting the aromatic rings were synthesised and evaluated for their antiproliferative activities against three human cancer cell lines (SGC-7901, A-549 and HT-1080) using an MTT assay. Compounds 10a, 10b, 7a, 7b and 7f showed significant activities against these cell lines. The most potent compound in this series, 10a, was selected to investigate its antitumour mechanism. In addition, molecular docking studies suggested that compound 10a interacts very closely with the nocodazole docking pose through hydrogen bonds at the colchicine binding site of tubulin.

Conditioned fear stress combined with single-prolonged stress: a new PTSD mouse model

There are still some defects in current single-prolonged stress (SPS) model and conditioned fear (CF) stress model of post-traumatic stress disorder (PTSD). The purpose of this study is to evaluate a novel mouse model of PTSD. Male KM mice suffered the double stresses-SPS and CF. After incubation time, the novel model exhibited the PTSD-like behaviors: sensitive fear and conditioned fear, low activities and defects in novel object recognition abilities. The apoptosis in the hippocampus was significantly increased, which was induced by the double stresses and further caused the synaptic structure damages in the hippocampus. The electron microscopy analysis further proved the synaptic losses and neuronal impairments in the hippocampus. Our results indicated this combined stresses mouse model was better than the SPS model and CF model. In addition, in order to further verify this model, paroxetine was administered after the double stresses. The results showed that paroxetine administration reduced PTSD-like behaviors, hippocampal apoptosis and structure damages. We conclude that this mouse model is novel and more predictably mimicked the clinical characteristics of PTSD, and this model can be further used for investigating the mechanisms of PTSD and screening effective therapeutics agents.

DAT-230, a novel microtubule inhibitor, exhibits potent anti-tumor activity by inducing G2/M phase arrest, apoptosis in vitro and perfusion decrease in vivo to HT-1080

PurposeThe anti-mitotic agent, combretastatin A-4 (CA-4), is the lead compound of a new class of anti-cancer drugs that target tumor vasculature. 2-Methoxy-5-(2-(3, 4, 5-trimethoxyphenyl) thiophen-3-yl) aniline (DAT-230) is a structurally novel CA-4 analog with more stability. We investigated its anti-tumor activity and mechanisms in vitro and in vivo for the first time.MethodsCytotoxicity was measured by MTT method. Apoptosis, mitochondria membrane potential (ΔΨm) and NO generation were measured by flow cytometry. Intracellular microtubule network was detected by immunofluorescence experiments. Protein expression was analyzed by Western blotting. In vivo, the anti-tumor activity was assessed using fibrosarcoma xenografts subcutaneously established in BALB/c nude mice. Vasculature perfusion was identified using fluorescent DNA-binding compound Hoechst 33342.ResultsDAT-230 exhibited potent anti-proliferative activity against various cancer cells. DAT-230-treatment in HT-1080 cells resulted in microtubule de-polymerization and G2/M phase arrest preceding apoptosis. Phosphor-cdc2 (thr14/tyr15) reduction, cyclin B1 accumulation and aberrant spindles denoted the cyclin B1-cdc2 complex active and M phase arrest in HT-1080 cells treated with DAT-230. Apoptosis induced by DAT-230 was related with the activation of caspase-9, caspase-3 and PARP cleavage, which were at the downstream of mitochondria. The decrease ratio of Bcl-2/Bax, elevation of NO and disruption of ΔΨm confirmed the causal relationship between DAT-230 and mitochondrial pathway. In vivo, DAT-230 delayed tumor growth, induced tumor perfusion decrease and extensive hemorrhagic-necrosis.ConclusionsDAT-230 is a promising microtubule inhibitor that has great potential for the treatment of fibrosarcoma in vitro and in vivo. Its potential to be a candidate of anti-cancer agent is worth being further investigated.

Berberine acutely inhibits the digestion of maltose in the intestine.

ETHNOPHARMACOLOGICAL RELEVANCEnThe Chinese Goldthread Rhizome has been used in the Traditional Chinese Medicine as an important ingredient of many formulas for the treatment of diabetes mellitus. Berberine, the main effective composition of Chinese Goldthread Rhizome, is also effective in treating diabetes in todays clinical practice of Traditional Chinese Medicine.nnnAIM OF THE STUDYnTo evaluate the hypoglycemic activity of berberine which treats acutely on the postprandial blood glucose, and to explore the mechanism of this activity.nnnMATERIALS AND METHODSn1. One-dose preprandial intragastric administrations of berberine were given to normal animals (dogs and rats), and the postprandial blood glucose concentration curves were measured. Serum insulin enzyme linked immunosorbent assay (ELISA) was only performed in rats. 2. The euglycemic clamp test was performed to evaluate the effect of one-dose berberine intragastric administration on the blood glucose transformation and utilization rate in rats. 3. In the Caco-2 cell monolayer test, the changes of glucose concentration on the apical and basolateral sides were measured when the maltose solution containing berberine was added to the apical side. 4. The inhibition ratio of berberine against α-glucosidase was measured in vitro. 5. The effect of berberine on the fluorescence emission spectrums of α-glucosidase was studied.nnnRESULTSnOne-dose preprandial intragastric administration of berberine delayed the rise of post-maltose blood glucose, did not affect postprandial blood glucose after glucose meal, and did not affect the insulin level in normal rats; reduced post-maltose blood glucose in normal dogs. 2. The result of euglycemic clamp test showed that one-dose intragastric administration of berberine had no effect on the blood glucose transformation and utilization rate in rats. 3. Berberine added to the maltose solution on the apical side of Caco-2 cell monolayer reduced the glucose concentration on the apical side. Glucose in basolateral side of all groups cannot be detected. 4. Berberine inhibited the activity of α-glucosidase in vitro. 5. Berberine significantly and concentration dependently quenched the fluorescence emission spectrum of α-glucosidase.nnnCONCLUSIONnOur findings suggest an additional mechanism of the hypoglycemic activity of berberine by demonstrating its ability to acutely inhibit the α-glucosidase, and support the traditional use of berberine and Chinese Goldthread Rhizome for the treatment of diabetes mellitus.

Effects of acute and chronic administration of MK-801 on c-Fos protein expression in mice brain regions implicated in schizophrenia with or without clozapine

This study investigated the effects of acute and chronic administration of the non-competitive NMDA receptor antagonists MK-801 on c-Fos protein expression in different brain regions of mice with or without clozapine. MK-801 (0.6 mg/kg) acute administration produced a significant increase in the expression of c-Fos protein in the layers III-IV of posterior cingulate and retrosplenial (PC/RS) cortex, which was consistent with the previous reports. Moreover, we presented a new finding that MK-801 (0.6 mg/kg) chronic administration for 8 days produced a significant increase of c-Fos protein expression in the PC/RS cortex, prefrontal cortex (PFC) and hypothalamus of mice. Among that, c-Fos protein expression in the PC/RS cortex of mice was most significant. Compared to acute administration, we found that MK-801 chronic administration significantly increased the expression of c-Fos protein in the PC/RS cortex, PFC and hypothalamus. Furthermore, pretreatment of mice with clozapine significantly decreased the expression of c-Fos protein induced by MK-801 acute and chronic administration. These results suggest that c-Fos protein, the marker of neuronal activation, might play an important role in the chronic pathophysiological process of schizophrenic model induced by NMDA receptor antagonist.

Existence of glia mitigated ketamine-induced neurotoxicity in neuron–glia mixed cultures of neonatal rat cortex and the glia-mediated protective effect of 2-PMPA

The present study compared ketamine-induced neurotoxicity in the neuron-glia mixed cultures and neuronal cultures and further explored the neuroprotective effect of the NAAG peptidase inhibitor 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Firstly, Rosenfelds staining and immunofluorescence staining of microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) were used to address the difference of morphology in the mixed cultures and neuronal cultures. Our results showed that neurons and astrocytes grew in good conditions. The ratio of neurons and astrocytes in the mixed cultures was around 1:1, and the purity of neurons in the neuronal cultures is 91.3%. Furthermore, ketamine was used to test the hypothesis that the presence of a higher proportion of glia in the mixed cultures would be protective against ketamine-induced neurotoxicity in the mixed cultures compared with neuronal cultures. The results showed that ketamine-induced morphological changes, cell viability decrease and lactate dehydrogenase (LDH) levels increase were significantly mitigated in neuron-glia mixed cultures compared with neuronal cultures. Furthermore, 2-PMPA was included to further explore efficient protective drug for ketamine-induced neurotoxicity. Our results showed that 2-PMPA reduced ketamine-induced decrease of cell viability and increase of LDH levels in the mixed cultures but not in the neuronal cultures. Further morphological changes of neurons and astrocytes also indicated that 2-PMPA could improve ketamine damaged neurons in the mixed cultures instead of neuronal cultures. These results indicate that glia protect neurons from ketamine-induced neurotoxicity. These data further suggest that glia mediate the neuroprotective effect of 2-PMPA and 2-PMPA has the potential to treat ketamine-induced neurotoxicity in vivo. Delineating the mechanisms underlying the communication between neurons and glia and the neuroprotective effects of 2-PMPA in the mixed cultures to ketamine-induced neurotoxicity require further investigation.