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OBJECTIVE To explore the neuro-protective effects of saffron (Crocus satius L.) on chronic focal cerebral ischemia in rats.METHODS SD rats were randomly divided into 6 groups:sham control group,MCAO group,edaravone group and saffron 30,100,300 mg·kg-1groups.Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).Saffron was administered orally by once daily from 2 h to 42 d after ischemia. At 42 d after cerebral ischemia, neurological deficit score, spontaneous activity test,elevated plus maze test,marble burying test and novel objective recognition test were used to evaluate the effects of saffron on the behevioural change. Infarct volume, survival neuron density, activated astrocyte number, and the thickness of glial scar were also detected. GFAP expression and inflammatory cytokine contents in ischemic peripheral region were detected by Western blot and ELISA,separately.RESULTS Saffron(100,300 mg·kg-1)improved the body weight decrease, neurological deficit and spontaneous activity. Saffron (30-300 mg·kg- 1) increased the traveled distance ratio and total time in open arm, decreased the buried marble number, which indicated that saffron could ameliorate anxiety- and depression-like behaviors. Saffron (100, 300 mg·kg-1)improved the learning and memory function,which manifested by increased discrimination ratio(DR)and discrim-ination index (DI) in T2test. The results of toluidine blue found saffron treatment (100, 300 mg·kg-1) decreased the infarct volume and increased the neuron density in cortex and hippocampal.The activated astrocyte number,the thickness of glial scar and GFAP expression in ischemic peripheral region decreased after saffron. Saffron (100, 300 mg·kg-1) decreased the contents of IL-6 and IL-1β, increased the content of IL-10 in ischemic peripheral region.CONCLUSION Saffron exerted neuro-protective effects on chronic focal cerebral ischemia,which could be related with inhibiting the activation of astrocyte and glial scar,following with the decrease of inflammatory reaction.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.</p><p><b>RESULTS</b>RP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).</p><p><b>CONCLUSIONS</b>HJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.</p>
Subject(s)
Animals , Rats , Berberine , Berberine Alkaloids , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Flavanones , Flavonoids , Glucose , Metabolism , Iridoids , Models, Animal , Neurons , Oxygen , Metabolism , Reperfusion Injury , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To investigate the efficacy of novel object recognition (NOR) test in assessment of learning and memory ability in ICR mice in different experimental conditions.</p><p><b>METHODS</b>One hundred and thirty male ICR mice were randomly divided into 10 groups: 4 groups for different inter-trial intervals (ITI: 10 min, 90 min, 4 h, 24 h), 4 groups for different object materials (wood-wood, plastic-plastic, plastic-wood, wood-plastic) and 2 groups for repeated test (measured once a day or every 3 days, totally three times in each group). The locomotor tracks in the open field were recorded. The amount of time spent exploring the novel and familiar objects, the discrimination ratio (DR) and the discrimination index (DI) were analyzed.</p><p><b>RESULTS</b>Compared with familiar object, DR and DI of novel object were both increased at ITI of 10 min and 90 min (P<0.01). Exploring time, DR and DI were greatly influenced by different object materials. DR and DI remained stable by using identical object material. NOR test could be done repeatedly in the same batch of mice.</p><p><b>CONCLUSION</b>NOR test can be used to assess the learning and memory ability in mice at shorter ITI and with identical material. It can be done repeatedly.</p>
Subject(s)
Animals , Male , Mice , Learning , Memory , Mice, Inbred ICR , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective effect of cilostazol administrated intranasally on chronic injury after focal cerebral ischemia in mice.</p><p><b>METHODS</b>Focal cerebral ischemia in mice was induced by middle cerebral artery occlusion (MCAO). Cilostazol was administrated intranasally or intraperitoneally 1 h, 4 h and 7 h after the operation; then twice a day from the second day for 2 weeks. The neurological deficit scoring and the inclined board testing were performed within 35 d after ischemia. The survival rate, infarct volume and neuron density were assessed 35 d after ischemia.</p><p><b>RESULT</b>Intranasal cilostazol at 0.3 mg/kg increased the survival rate. Intranasal cilostazol (0.3 mg/kg, 1 mg/kg) and intraperitoneal cilostazol (10 mg/kg) significantly attenuated neurological deficit, reduced infarct volume, and increased the survival neuron density in the border of ischemia region.</p><p><b>CONCLUSION</b>Cilostazol administered intranasally demonstrates protective effects on chronic cerebral ischemia in mice.</p>
Subject(s)
Animals , Male , Mice , Administration, Intranasal , Brain , Pathology , Brain Ischemia , Drug Therapy , Pathology , Disease Models, Animal , Infarction, Middle Cerebral Artery , Drug Therapy , Pathology , Neurons , Pathology , Tetrazoles , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the neuroprotective effects of Chinese herb medicine Huanglian-Jiedu-Tang (HJDT) on chronic brain injury after focal cerebral ischemia in mice.</p><p><b>METHODS</b>Focal cerebral ischemia was induced by occlusion of right middle cerebral artery (MCA) for 15 min. HJDT (at dosage of 2 g/kg or 4 g/kg, qd, orally) was administered for 21 d from d 7 before ischemia until d 14 after ischemia. The sham and ischemic controls were administered with normal saline orally. The neurological deficit scoring and the inclined board testing were performed within 35 d after ischemia. The survival rate, the infarct volume and the neuron density were assessed 35 d after ischemia.</p><p><b>RESULT</b>HJDT increased the survival rate at dose of 4 g/kg; significantly reduced the neurological deficits, infarct volume and cerebral atrophy at doses of 2 and 4 g/kg after ischemia; and significantly increased the neuron density in the ischemic hippocampal CA1 region, striatum and cortex at dose of 4 g/kg but only increase the density in hippocampal CA1 region at dose of 2 g/kg.</p><p><b>CONCLUSION</b>Chinese herb medicine HJDT has neuroprotective effects on chronic brain injury after focal cerebral ischemia in mice.</p>
Subject(s)
Animals , Male , Mice , Behavior, Animal , Physiology , Brain , Pathology , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Infarction, Middle Cerebral Artery , Drug Therapy , Pathology , Neurons , Pathology , Neuroprotective Agents , Pharmacology , Therapeutic Uses , PhytotherapyABSTRACT
<p><b>OBJECTIVE</b>To develop a novel method for continuously assessing the spatio-temporal properties of locomotor activity of mice in an open field using a video-tracking system.</p><p><b>METHODS</b>The locomotor tracks in the open field were recorded by video camera within 22 h, and analyzed by AnalyPower1.1 system that we developed recently. Total distance, distances traveled in different zones and their ratios to total distance; total time,times spent in different zones and their ratios to total time were used as indicators to assess the properties of locomotor activity.</p><p><b>RESULTS</b>In free and wakeful state, the locomotor activity of mice presented obvious regional and temporal properties. Mice preferred to stay in home base (food and water zones), and frequently visited the peripheral zones but seldom the center zones within 22 h. On the other hand, mice were most active within the first 1 h, and then their activity obviously decreased. After their activity became stable, the mice showed the obvious circadian variation of the activity as they were more active in the night.</p><p><b>CONCLUSION</b>The novel method we developed in this study can continuously assess the spatio-temporal properties of locomotor activity quantitatively and objectively.</p>
Subject(s)
Animals , Male , Mice , Behavior, Animal , Physiology , Circadian Rhythm , Physiology , Environment , Exploratory Behavior , Physiology , Locomotion , Physiology , Motor Activity , Physiology , Time Factors , Video RecordingABSTRACT
The purpose of this study was to develop a quantitative and objective method for evaluating neurological deficits in mice with focal cerebral ischemia. After middle cerebral artery occlusion (MCAO), the neurological deficits were evaluated 24 h later. We measured the mean angles, dominant angles and turns in a hanged test in which the mice were sticked on the wall, and the holding angles in an inclined plane test as well, Then we determined the cerebral infarct volumes, neuron density in hippocampus, cortex and subcortical areas 24 h after MCAO. The correlations among infarct volume, neuron density and neurological deficits were analyzed. We also compared the quantitative method with two typical complex methods of behavioral assessment. The effect of [pranlukast, 4-oxo-8-[p-(4-phenylbutyloxy) benzoylamino]-2-(tetrazol-5-yl)-4H-1-benzopyran hemihydrate] (ONO-1078), a neuroprotective agent, on ischemic injury was observed using this method. We found that the variables measured by both quantitative and typical behavioral methods significantly changed in the ischemic mice, and correlated with the infarct volumes and neuron densities. The quantitative variables well correlated with those of typical behavioral assessment, too. ONO-1078 inhibited ischemic injury and reduced the total scores of quantitative assessment. Thus, the quantitative method we developed is useful in evaluating neurological deficits of focal cerebral ischemia with the advantages of objectivity, quantification, simplicity and non-invasion, and can be used in the evaluation of neuroprotective effects of drugs.
Subject(s)
Animals , Female , Male , Mice , Behavior, Animal , Brain , Pathology , Brain Ischemia , Pathology , Chromones , Pharmacology , Therapeutic Uses , Hippocampus , Pathology , Infarction, Middle Cerebral Artery , Pathology , Leukotriene Antagonists , Pharmacology , Therapeutic Uses , Mice, Inbred ICR , Neurologic Examination , Neuroprotective Agents , Pharmacology , Therapeutic UsesABSTRACT
OBJECTIVE: To evaluate the feasibility of light transmission to measure focal cerebral ischemia in mice. METHODS: Persistent focal cerebral ischemia was induced by middle cerebral artey occlusion (MCAO) in mice. The brain were removed 24 h after MCAO and coronally dissected into 1 mm sections. Using a stereomicroscope, the brain section was illuminated with a halogen lamp and computerized images were stored. Next the brain sections were stained for 30 minutes with 0.5% TTC (2, 3, 5-triphenylterzolim chloride) at 37 degrees C. Using an image analyzer (AnalyPower 1.0), the infarct volumes obtained by light transmittance and TTC staining were calculated. Integrated gray scales of sections of both hemispheres were calculated by Photoshop 5.0. RESULTS: A close correlation existed between cerebral infarct volume measured by light transmission and TTC staining (r=0.81). The mean gray scales measured by both techniques of the ischemic hemispheres as well as those of the cortex, subcortex and hippocampus were siginificantly higher than those of non-ischemic hemispheres and of control mouse hemispheres (P <0.001). Further there were no significant difference between the two hemispheres of control mice and between hemispheres of control mice and non-ischemic hemispheres of the MCAO mice. CONCLUSION: Light transmission can be used for qualitative analysis of focal cerebral ischemia.