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Objective To observe the expressions of NeuN and pCaMKⅡα in brain and test the spacial learning and memory in neonatal hypoxic?ischemia encephalopathy ( HIE ) model mice. Methods 7d ICR mice were randomly divided into sham group( n=19) and model group( n=23). HIE model was induced by right common carotid artery ligation followed by 8% oxygen hypoxia for 100 min. DAPI staining was used to examine brain pathological change,immunofluorescent staining was used to examine the expression of NeuN and pCaMKⅡα in the ipsilateral brain,and Morris water maze was used to test the spa?cial learning and memory. Results Mice in sham group showed that brain cells were arranged in a dense and orderly manner,the number of NeuN?positive cells and pCaMKⅡα?positive cells were (106.50±20.07), (87.17±16.55) respectively in the brain,and the escape latency was short. Compared with mice in sham group,mice in model group showed more cells loss,less NeuN?positive cells(19.17±3.60) and less pCaMKⅡα?positive cells(13.33±3.62) in the ipsilateral hemisphere,and longer escape latency(P<0.01). Conclu-sion The spacial learning and memory are impaired in hypoxia ischemia,which may be related to the de?creasing expression of pCaMKⅡα in neurons in ipsilateral brain.
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Objective To observe apoptotic cells and caspase-3-positive cells in ipsilateral neonatal hypoxic-isch?emia encephalopathy (NHIE) model in mice. Methods CD1 mice of age 7 days (n=30) were randomly divided into two groups: sham group (n=9) and model group (n=21). NHIE model was induced by right common carotid artery ligation fol?lowed by 8%oxygen hypoxia for 100 min. TTC staining was used to determine area of brain infarction. DAPI staining was used to detect pathological change in brains. TUNEL assay was used to detect apoptotic cells and fluorescence immunohisto?chemistry was used to detect caspase-3 expression in the ipsilateral brain. Results No infarct was detected in sham group. Cells were densely and orderly arranged in brain. TUNEL-positive cells (18.57±4.98) and caspase-3-positive cells (9.17± 2.14) in the ipsilateral brain were both less than those in the ipsilateral brain of mice in model group (209.57±41.27) and (63.33±16.22) respectively. Mice in model group presented infarct in the right hemisphere with more dead cells and wider in?terstitial space compared with sham group. Conclusion Brain injury in NHIE model might be related to the increasing cas?pase-3 expression thus leads to apoptosis.
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Objective To observe the effect of moxibustion pretreatment on caspase-3 expression in cortex of rats with cerebral ischemia-reperfusion injury. Methods Twenty-one male SD rats were randomly divided into three groups, sham group (n=6), model group (n=8) and moxibustion pretreatment group (n=7). Focal cerebral ischemia-reperfusion injury rat model was induced by middle cerebral artery occlusion (MCAO) for 2 hours followed by reperfusion. Brain infarct was ex-amined by TTC staining. Brain morphology was shown by HE staining and the ultrastructure of cell was observed with elec-tron microscope. Caspase-3 expression in cortex was examined by immunofluorescence histochemistry. Results The per-centage of infarct volume [(24.9±4.7)%] in moxibustion pretreatment group was much smaller than that [(45.8±4.6)%] in mod-el group. Nerve cells in moxibustion pretreatment group experienced less morphological and structural changes than those in model group, and caspase-3 positive cells (39.17 ± 7.28) in moxibustion pretreatment group were less than those (58.17 ± 16.53) in model group. Conclusion Moxibustion pretreatment could attenuate cerebral ischemia-reperfusion injury, which might be through decreasing caspase-3 expression.
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Objective To observe the effects of heat-sensitive moxibustion on the expression and activity of super-oxide dismutase (SOD) and the content of malondialdehyde (MDA) in focal cerebral ischemia-reperfusion injury in rats, and the mechanism thereof. Methods Thirty-six male SD rats were randomly divided into four groups, sham-operated group (n=6), ischemia-reperfusion (I/R) injury group (n=10), I/R injury with 15-minute moxibustion group (n=10) and I/R injury with 35-minute moxibustion group (n=10). The focal cerebral ischemia-reperfusion rat model was induced by middle cere-bral artery occlusion (MCAO) for 2 h followed by reperfusion. Values of SOD activity and MDA content were determined by colorimetry, and the cortical expression of SOD2 protein was detected by Western blot technique. Results Values of SOD activities were significantly higher in serum (22.78±1.31)U/mL and cortex (4 909.6±1 345.6) U/g of heat-sensitive moxibus-tion group than those of model group (20.17±1.12)U/mL and (2 602.0±1 515.5)U/g. Values of MDA contents were significant-ly decreased in serum (3.78±2.00)μmol/L and cortex (1 226.5±38.4)nmol/g in heat-sensitive moxibustion group than those of model group (16.82 ± 6.70)μmol/L and (1 905.6 ± 478.6) nmol/g. The cortical expression of SOD2 protein (0.974 ± 0.166) was higher in heat-sensitive moxibustion group than that of model group (0.702±0.040). Conclusion Heat-sensitive moxi-bustion could reduce the damage of cerebral inchemia-reperfusion, which might be through improving SOD activity, increas-ing SOD expression and decreasing MDA content.
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ObjectiveTo investigate the effects of moxibustion on the expression of tryptophan hydroxylase 2(TPH2) mRNA in median raphe nuclei and the activity of monoamine oxidase (MAO) in serum and cortex in depression model rats and to explore its mechanism.MethodsFemale SD rats were randomly divided into control group(n=6),model group(n=7) and moxibustion group(n=7).The rat depression model was induced by giving isolation-housing in combination with chronic unexpected mild stress(CUMS) for 21 days.The hemi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) was employed to detect the relative expression of TPH2 mRNA,colorimetry was used to test the activity of MAO.ResultsThe expression of TPH2 mRNA in median raphe nuclei in rats of the moxibustion group was higher than that in the model group( relative dentisty value:(0.3759 ± 0.0272)vs(0.2573 ± 0.0581 ),P ＜ 0.05 ).The activity of MAO in serum and cortex was reduced in rats of the moxibustion group( ( 1.919 ± 1.243 ) U/ml,( 1292.048 ± 90.072 ) U/mg protein,respectively) compared with that in the model group ( (4.117 ± 2.727 ) U/ml,( 1441.345 ± 117.050 ) U/mg protein,respectively).ConclusionMoxibustion has the effects of improving depression,increasing the expression of TPH2 mRNA in median raphe nuclei and reducing the activity of MAO in serum and cortex might be one of its mechanisms.
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Objective To observe the effect of Chinese medicine of replenishing the liver and kidney on the concentration of 5-hydroxytryptamine(5-HT)in hippocampus and the expression of tryptophan hydroxylase 2 (TPH2)in median raphe nuclei in perimenopausal depression model rats and to explore its mechanism.Methods The animal model was established by resecting the ovaries of female rats,then giving isolation-housing in combination with 21 days of chronic unexpected mild stress(CUMS).High performance liquid chromatogram-electrochemical detection(HPLC-ECD)were employed to measure 5-HT in hippocampus,the hemi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)and fluorescence immunohistochemistry were employed to detect the relative expression of TPH2 mRNA and number of TPH2 immunoreactive neurons in median raphe nuclei.Results The concentration of 5-HT in hippocampus was increased in rats of the medicine group compared with that in the model group((2543.06±859.59)ng/g tissue,(1845.81±233.55)ng/g tissue,F=9.617,P＞0.05).RT-PCR showed that the relative expression of TPH2 mRNA in medicine group was much higher than that in model group ((1.282±0.158),(0.985±0.120),F=3.552,P＜0.05).Immunofluorescence showed that the number of TPH2 immunoreaetive neurons in raphe nuclei had statistical significance between medicine group and model group ((152.74±68.52),(74.12±38.01),F=7.939,P＜0.01).Conclusion Chinese medicine replenishing liver and kidney have effects of improving perimenopausal depression,increasing the expression of TPH2 in median raphe nuclei,which leads to enhance the synthesis of 5-HT in hippocampus might be one of its mechanisms.
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Objective To observe changes of spacial learning and memory and the ultrastructure of hippocampus in perimenopausal depression model rats. Methods Thirty-six female SD rats were randomly divided into four groups(9 in each). Normal group wasn't given any stress, and stressed group was given isolation-housing in combination with chronic unexpected mild stress (CUMS) for 21 days. Ovariectomized group was given ovary resecting, and model group was made by resecting the ovaries of female rats, then giving isolation-housing in combination with 21 days of CUMS. Morris water maze test were used to test rats' behaviors. Light microscope and transmission electron microscope were used to observe the tissular structure of hippocampus. Results After ovaries resecting and 21 days of CUMS,the escape periods of rats in model group during the first three days ( (44.08 ± 18.29)s; ( 38.80 ± 19.07 ) s; ( 22.74 ± 14.13 ) s) were longer than ovariectomizd group ( (43.68 ± 10. 37 ) s; ( 20.28 ±17.75)s;(21.22 ± 11.91 )s] and normal group( (34.50 ± 11.08)s;(21.42 ± 14.32) s;( 13.94 ±9.76) s). HE staining showed that hippocampal pyramidal cell layer became thinner in rat model of depression,the gap between cells increased,and the cells arrayed in disorder,or loosely,even a large number of cells were lost. Transmission electron microscope showed that in rats of model group, hippocampal neurons were less, nuclear collapsed, the whole nuclear membrane was fuzzy and there were some local fractures in the nuclear membrane. Mitochondria within the cytoplasm were swelling and their ridges disappeared. Conclusion The decline of spacial learning and memory abilities in perimenopausal depression model rats might result from pathological changes in hippocampus.