Effect of BDNF expression in cerebral cortex and hippocampus on ability of learning and memory in APP/PS1 transgenic mice

AIM: To investigate the expression changes of brain-derived neurotrophic factor (BDNF) in the cerebral cortex and hippocampus and their effects on the ability of learning and memory in the wild-type (WT) mice and APP/PS1 transgenic mice. METHODS: WT mice and APP/PS1 transgenic mice were selected as study subjects. Aβ plaques, apoptosis rate and BDNF expression in the cerebral cortex and hippocampus of WT mice and APP/PS1 transgenic mice were detected by the methods of Congo red staining, TUNEL, immunofluorescence and Western blot. The abilities of learning and memory were determined by Morris water maze test. RESULTS: The Aβ plaques appeared in the cerebral cortex and hippocampus of APP/PS1 transgenic mice, and the number of Aβ plaques in 12-month-old mice was larger than that in 6-month-old mice (P<0.05). The number of apoptotic neurons in the cerebral cortex and hippocampus of 12-month-old APP/PS1 transgenic mice was larger than that of WT mice (P<0.01). The expression level of BDNF in the cerebral cortex and hippocampus of WT mice was higher than that of APP/PS1 transgenic mice (P<0.01). The Morris water maze test showed that the escape latency in APP/PS1 transgenic mice was longer than that in WT mice, and the times across the platform quadrant in 60 s was less than that in WT mice (P<0.01). The swim-tracking path of APP/PS1 transgenic mice was disordered and irregular. CONCLUSION: The expression of BDNF in the cerebral cortex and hippocampus of APP/PS1 transgenic mice was lower than that of WT mice, accompanied by increased neuronal apoptosis and decreased spatial learning and memory ability. The decrease in learning and memory ability may be related to decreased BDNF expression in the cerebral cortex and hippocampus of APP/PS1 transgenic mice, leading to increased neuronal apoptosis, which may be one of the pathological mechanisms of Alzheimer disease.     

Flavonoids from stem and leaf of Scutellaria baicalonsis Georgi inhibit PHF abnormality and regulatory mechanism of protein phosphatase in rats' brain induced by okadaic acid

AIM: To investigate the effect of flavonoids from stem and leaf of Scutellaria baicalonsis Georgi (SSF) on paired helical filament (PHF) abnormality and the regulatory mechanism of protein phosphatase (PP) in rats' brain induced by okadaic acid (OA). METHODS: Male Sprague-Dawley (SD) rats were microinjected with OA (200 ng/kg) by the lateral ventricle to establish a memory impairment model. Morris water maze was used to screen the memory impairment model. The successful model rats were continuous intragastric infusion (ig) SSF for 36 days. The relative protein expression of PHF, PP1, PP2A-Cα, PP2A-Cβ, PP2CA and PP2CB in the rat cerebral cortex and hippocampus were detected by Western blot. GinKgo biloba leaf flavonoids (GLF) were used as positive control drug. RESULTS: Compared with the sham-operated rats, the relative protein expression of PHF in the cerebral cortex and hippocampus and PP1 in cortex of model rats were significantly increased (P<0.01), and the protein expression of PP2A-Cα, PP2A-Cβ in the cerebral cortex and hippocampus and PP2CB in the hippocampus were decreased (P<0.05), while the relative protein expression of PP2CA and PP2CB in the cortex were significantly increased (P<0.01). SSF reversed the abnormality in the protein expression of PHF, PP2A-Cα and PP2A-Cβ in rat cortex and hippocampus and PP1 in rat cortex induced by OA (P<0.01), which had no significant effect on the relative protein expression of PP2CA and PP2CB. GLF also showed similar results to SSF. CONCLUSION: SSF significantly reduces the abnormal formation of PHF in rats' brain induced by OA, which may be related to the regulation of PP1, PP2A-Cα and PP2A-Cβ expression, but not with PP2CA and PP2CB expression.     

Influences of Vit-E on the mtDNA damage and Ca2+ homostasis of hippocampus and antioxidative ability in brain of aging mice induced by D-galactose

AIM: To study the influences of vitamin E (Vit-E) on the mtDNA damage and Ca²⁺ homeostasis in hippocampus and antioxidative ability in aging brain induced by D-galactose.METHODS: D-galactose (1 000 mg·k^-1·d^-1 ) was injected into mice hypodermically for 8 weeks to induce aging animal model, and Vit-E (100 mg·kg^-1; 250 mg·kg^-1) was administered for 6 weeks by ig at the 3rd week of making model. After Vit-E treatment for 8 weeks, water maze test was used to determine the ability of mice’s learning and memory. The activities of glutathione peroxidase (GSH-Px) and succinate dehydrogenase (SDH), the content of nitric oxide (NO) and activity of nitric oxide synthase (NOS) in the brain tissue were detected separately. Fura-2/AM, double-wave-length fluorospectrophotometer and PCR method were used to measure the concentration of calcium ion and mtDNA mutation in the hippocampus cells.RESULTS: Administration of Vit-E improved significantly the ability of learning and memory in model mice, inhibited the activity of NOS and decreased the amount of NO, and increased the activities of GSH-Px and SDH respectively in brain tissues, decreased the concentration of calcium ion (P<0.01, P<0.05), and prevented the damage of mtDNA in hippocampus.CONCLUSION: Vit-E can enhance the antioxidative ability, regulate the homeostasis of Ca²⁺ and inhibit the damage of mtDNA caused by oxidative stress in aging brain, and improve the ability of learning and memory in aging mice.     

Effects of rolipram on learning and memory and the activity of PDE4 in hippocampus following the focal brain injury induced by ischemia- reperfusion in rats

AIM: To investigate the effects of rolipram on the ability of learning and memory and the activity of PDE4 in hippocampus following the focal brain injury induced by ischemia- reperfusion in rats. METHODS: The cerebral ischemia-reperfusion injury model was made by middle cerebral artery occlusion (MCAO) in rats. The rats were randomly divided into sham-operated group, model group, and rolipram group. Rolipram was administered once a day (1 mg/kg, ip) from 6 h after the onset of the operation for 2 weeks. Then the learning and memory abilities were tested after Morris water maze and step-though training. The activity of PDE4 in hippocampus was evaluated by HPLC. RESULTS: In the Morris water maze test, compared to sham-operated group, the platform-finding time and swimming distance in model group were significantly longer (P<0.05). Compared to model group, the platform-finding time and swimming distance in rolipram group were significantly shorter (P<0.05). In the step-through test, compared to sham-operated group, the lantent period in model group was significantly shorter (P<0.01) and the error times were statistically increased(P<0.05). Compared with model group, the lantent period in rolipram group were significantly longer (P<0.05), and the error times were markedly decreased. The assay of the HPLC demonstrated that the activities of PDE4 in hippocampus in model group were higher than those in the sham-operated group and rolipram group. CONCLUSION: Rolipram reduces the activity of PDE4 in hippocampus and enhances the ability of learning and memory after the injury induced by ischemia-reperfusion.     

Expression and histogenesis of matrix metalloproteinase-9 and transforming growth factor-beta 1 in acute cerebral ischemia model of rats

AIM:To observe the expression and tissue localization of matrix metalloproteinase 9 (MMP-9) and transforming growth factor beta 1 (TGF-β1) in the rat acute cerebral ischemia model. METHODS:Male Wistar rats were used to establish acute cerebral ischemia model by a suturing method. The rats were divided into normal control group, sham group and ischemia 6 h, 12 h, 1 d, 2 d, 6 d and 14 d groups. The rat cerebral cortex and hippocampus of the brain were collected at different time points.The mRNA and protein levels of MMP-9 and TGF-β1 in the brain tissues were detected by real-time PCR and in situhistochemistry staining, respectively. The levels of MMP-9 and TGF-β1 in the plasma were also measured by ELISA. RESULTS:The results of real-time PCR showed that the mRNA levels of MMP-9 began to increase 6 h after acute ischemia and reached to a peak 2 d after acute ischemia. Similarly, the mRNA level of TGF-β1began to rise 12 h after acute ischemia and reached to the highest level 6 d after acute ischemia. Compared with the sham rats, the mRNA levels of MMP-9 and TGF-β1 in the rat brains that collected at ischemic time of 12 h, 1 d, 2 d, 6 d and 14 d were significantly increased. Moreover, results of in situhistochemical staining showed that the expression of MMP-9 was detected at cerebral cortex and hippocampus 1 d after acute cerebral ischemia.Further studies showed that MMP-9 dyeing of the rat cerebral cortex was most obvious 2 d after the acute cerebral ischemia. Similarly, the rat cortex and hippocampus began to express TGF-β1 2 d after acute ischemia and TGF-β1 staining at rat cerebral cortex was most obvious 6 d after the acute cerebral ischemia. In addition, ELISA showed that the increase in MMP-9 and TGF-β1 was detected in the plasma 12 h after ischemia. Compared with the sham rats, the level of these 2 factors significantly upregulated since 1 d after ischemia. CONCLUSION: The brain tissue itself contributes to the upregulation of MMP-9 and TGF-β1 post acute cerebral ischemia, which shed light on the related research in the field.     

Effects of clonidine on learning and memory and protein levels of ERK signal pathway-related proteins in rats with chronic cerebral ischemia

AIM:To investigate the regulatory effects and underlying molecule-mechanism of clonidine on learning and memory in rats with chronic cerebral ischemia. METHODS:Sprague-Dawley rats (n=45) were randomly divided into sham-operation group, cerebral ischemia model group and clonidine group, 15 rats in each group. The chronic cerebral ischemia rat model was established by right middle cerebral artery occlusion for 2 h and reperfusion for 30 d. Clonidine was administrated by i.g. for 7 days in clonidine group. The ability of spatial reference memory of the rats with cerebral ischemia was tested by Morris water maze. The protein levels of extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), cAMP-response element binding protein (CREB) and phosphorylated CREB (p-CREB) were determined by immunohistochemistry and Western blot. RESULTS:The results of Morris water maze test showed that compared with the sham-operation group, the ability of spatial reference memory was obviously impaired in the cerebral ischemia model group. Compared with the cerebral ischemia model group, the ability of spatial reference memory in the clonidine group were improved. Compared with the sham-operation group, the protein levels of p-ERK1/2 and p-CREB in hippocampus were increased in model group (P<0.01). Compared with the cerebral ischemia model group, the protein levels of p-ERK1/2 and p-CREB in hippocampus were decreased in the clonidine group (P<0.01). CONCLUSION:Clonidine improves the learning and memory abilities of the rats with cerebral ischemia, and ERK1/2 and CREB are involved in this process.     

Effects of flavonoids isolated from Scutellaria stem and leaf on expression of NMDAR and VEGF in chronic cerebral ischemia rats

AIM: To study the effects of flavonoids isolated from Scutellaria stem and leaf (SSF) on the expression of N-methyl-D-aspartate receptor (NMDAR) and vascular endothelial growth factor (VEGF) in chronic cerebral ischemia rats. METHODS: The model of chronic cerebral ischemia was established by bilateral carotid artery occlusion for 2 months in female SD rats. The effects of SSF on mRNA expression of NMDAR in hippocampus and VEGF in cerebral cortex were evaluated by the method of RT-PCR. RESULTS: Compared with the sham group, the expression of NMDAR1, NMDAR2A and NMDAR2B in hippocampus and VEGF in cerebral cortex were significantly increased (P<0.01). However, the cerebral ischemia rats daily and orally administered with SSF at doses of 17.5 mg·kg^-1·d^-1, 35 mg·kg^-1·d^-1 and 70 mg·kg^-1·d^-1 for 38 days appeared that the mRNA expression of NMDAR1, NMDAR2A and NMDAR2B in hippocampus was obviously reduced (P<0.05), and the mRNA content of VEGF in the cortex (P<0.05) was increased. CONCLUSION: SSF decreases the expression of NMDAR in hippocampus, increases the expression of VEGF in cerebral cortex of cerebral ischemia rats, suggesting that the neuroprotective effect of SSF may be exerted by influencing the production of NMDAR and VEGF in the brain.     

Intervention of gossypol and relationship between cognitive function and expressions of 11β-HSD1 and GR in brain of type 2 diabetic rats

AIM: To study the effect of gossypol on the cognitive function of type 2 diabetic rats, and to explore its mechanism. METHODS: Thirty male Sprague-Dawley rats were divided into three groups randomly: normal group, type 2 diabetic group and gossypol treated group. After fed with high-fat diet for 4 weeks, the later two groups were injected with streptozotocin intraperitoneally to establish type 2 diabetic rat model. The animals in gossypol treated group were given gossypol at dosage of 15 mg/kg once per day for 4 weeks by gavage. Since 5th week, the times of gavages were changed into once per week at the same dosage and lasted to 12th week. Learning and memory abilities of rats were assayed with Morris water maze test. The concentration of blood glucose was measured by biochemical method. The levels of serum corticosterone and insulin were detected by enzyme-linked immunosorbent assay and radioimmunoassay, respectively. The protein expressions of 11β-HSD1 and GR in cerebral cortex and hippocampus were determined by Western blotting. The morphological changes of cerebral cortex and hippocampus were observed under light microscope and transmission electronic microscope, respectively. RESULTS: Compared to normal group, the karyopyknosis, dilation of golgiosome and mitochondria swelling of neuron from cerebral cortex and hippocampus were prominent in diabetic group. The concentrations of blood glucose, serum corticosterone and insulin increased significantly (P<0.01). Protein expression of GR decreased (P<0.05), 11β-HSD1 protein tended to increase. Platform searching score was lower (P<0.01) and escape latency was longer (P<0.01) in diabetic group. After treated with gossypol, the concentrations of blood glucose, serum corticosterone and insulin declined (P<0.01). The protein expression of 11β-HSD1 was decreased (P<0.05) and GR was increased (P<0.05). Escape latency was shorter (P<0.01) and platform searching score was increased (P<0.01). CONCLUSION: Gossypol may improve the cognitive function of type 2 diabetic rats. Decreasing the level of 11β-HSD1 and increasing GR protein in the brain may be involved in the mechanism.     

Effect of Buyanghuanwu decoction on the expression of ERK2 and CaMKⅡβ in hippocampus tissue and on ability of learning and memory in vascular dementia rats

AIM: To investigate the effect of Buyanghuanwu decoction, a Chinese medicine, on the ability of learning and memory in the rats with vascular dementia (VD) and on the protein expression of extracellular signal-regulated kinase 2(ERK2) and calcium/calmodulin-dependent protein kinase Ⅱβ(CaMKⅡβ) in hippocampus CA1 area.METHODS: The rats were divided into 4 groups: sham group, VD group, VD+Buyanghuanwu decoction group and VD+nimodipine group. The VD rat model was prepared by Pulsinelli's four-vessel occlusion. At 7th day, 14th day or 28th day after operation, the behaviors of the rats were tested by Morris water maze. The morphological changes of the neurons in hippocampus CA1 area were observed by HE staining 30 d after operation. Western blotting was used to observe the protein expression of ERK2 and CaMKⅡβ in the brain tissues of hippocampal CA1 area of the VD rats. RESULTS: Compared with sham group, the pathological changes such as irregular arrangement, coagulation necrosis and obvious deletion in the neurons of hippocampus CA1 area in VD group appeared significantly. The obstacle of learning and memory ability was observed and the protein expression of ERK2 and CaMKⅡβ in hippocampal CA1 area was significantly decreased (P<0.05). Compared with VD group, the neurons in hippocampal CA1 area of VD+Buyanghuanwu decoction group and VD+nimodipine group were in eumorphism, lined up in order, and the structure was close to that in sham group. The ability of learning and memory also significantly improved (P<0.05). The protein expression of ERK2 and CaMKⅡβ in hippocampal CA1 area significantly increased (P<0.05). CONCLUSION: Buyanghuanwu decoction promotes the protein expression of ERK2 and CaMKⅡβ in hippocampus CA1 area to protect the neurons from injury, builds up the synapses and promotes the ability of learning and memory in VD rats.     

Study of zinc element and ZnT3 mRNA expression in mouse brain

AIM:To explore the interaction and function between ZnT3 mRNA expression and zinc elemental distribution in brain slice of mouse.METHODS:Zinc distribution was determined by SRXRF and ZnT3 mRNA expression in tissue was examined by RT-PCR method.RESULTS:Zinc content in cerebral cortex and hippocampus was significantly higher than that in other positions.The highest expression of ZnT3 mRNA was observed in cerebrum,hippocampus and testis.However,the ZnT3 mRNA was not detected in heart,liver,lung,spleen,kidney,intestine,olfactory bulb and cerebellum.CONCLUSION:ZnT3 facilitates the accumulation of zinc in synaptic vesicles and may play important roles in structuring of vesicular zinc pool.     

Scutellaria barbata flavonoids inhibits NFT aggregation and regulatory mechanism in rats induced by composited Aβ

AIM: To investigate the effects of Scutellaria barbata flavonoids (SBF) on neurofibrillary tangle (NFT) aggregation, tau protein phosphorylation and the regulated mechanism of glycogen synthase kinase (GSK) 3β and protein phosphatase (PP) 2A in the rats induced by amyloid β protein 25-35 (Aβ_25-35) in combination with AlCl₃ and recombinant human transforming growth factor (RHTGF)-β1(composited Aβ). METHODS: The male SD rats were used to establish the simulated Alzheimer disease (AD) model by intracerebroventricular injection of composited Aβ. The Morris water maze was applied for screening the successful model rats with learning and memory deficits. The successful model rats were daily and orally administrated with SBF at doses of 35, 70 and 140 mg/kg or positive control drug Ginkgo biloba leaves flavonoids (GLF) at 140 mg/kg for 37 d. The silver nitrate staining was used to determine the cortical NFT. The protein levels of total tau, phosphorylated protein of tau at Ser199 and Ser214 sites, GSK3β and PP2A in hippocampus and cortex were determined by Western blot. The mRNA expression of GSK3β and PP2A in the hippocampus and cortex was detected by RT-PCR. RESULTS: Compared with sham group, the cell number of positive NFT with silver nitrate staining in model rat cerebral cortex was significantly increased. The protein levels of phosphorylated tau protein at Ser199 and Ser214 sites, GSK3β in the hippocampus and cerebral cortex in the model rats dramatically elevated, and PP2A was marked decreased as compared with the sham group rats. Meanwhile, the mRNA expression of GSK-3β significantly increased but PP2A was decreased. However, these above abnormalities were differently attenuated by treating with SBF at different doses or GLF at 140 mg/kg for 37 d. CONCLUSION: SBF suppresses the NFT aggregation by inhibition of the regulatory functions of GSK-3β and PP2A, thus reducing the phosphorylation of tau protein.     

Effect of microRNA-155 on regulation of angiogenesis in diabetic rats with cerebral ischemic injury

AIM: To evaluate the effect of microRNA-155(miRNA-155) on the regulation of angiogenesis in diabetic rats with cerebral ischemic injury. METHODS: Adult male Sprague-Dawley rats were randomly divided into 5 groups:sham group, cerebral ischemia group, diabetic cerebral ischemia group, diabetic cerebral ischemia+miRNA-155 inhibitors group and diabetic cerebral ischemia+scramble group. Diabetes model was made by injection of streptozocin and permanent cerebral ischemic model was developed by suture-occluded method. The scores of neurological deficit and infarct volume were estimated at 24 h after cerebral ischemia. miRNA-155 level was detected by real-time polymerase chain reaction. The expression of platelet endothelial cell adhesion molecule-1(PECAM-1/CD31) and vascular endothelial growth factor(VEGF) was detected by Western blotting. RESULTS: miRNA-155 inhibitor significantly reduced miRNA-155 levels in the ischemic cortex(P<0.05), improved the scores of neurological deficit, reduced infarction size and upregulated the levels of CD31 and VEGF(P<0.05). CONCLUSION: miRNA-155 has a critical role in the regulation of angiogenesis in diabetic rats with cerebral ischemia. Down-regulation of miRNA-155 using miRNA-155 inhibitor attenuates brain infarct injury in diabetic rats.     

Effect of ginsenoside Rg1 on activity and protein expression of NOS in rats with cerebral ischemic reperfusion injury

AIM: To observe the neuroprotective effects of ginsenoside Rg1 on focal cerebral ischemia reperfusion (I/R) injury in rats. METHODS: SD rats were applied to right middle cerebral artery occlusion (MCAO) for 2 h followed by 24 h of reperfusion. The rats were randomly divided into sham-operation group, I/R group and ginsenoside Rg1 pretreatment groups. The rats in ginsenoside Rg1 pretreatment groups were pretreated with ginsenoside Rg1 at doses of 10, 20 or 40 mg/kg once a day for 7 days and then subject to MCAO. The neurological deficit score was measured by Longa's method. The neurons were observed with Nissel staining. The nitric oxide (NO) content, the activity of nitric oxide synthase (NOS) and inducible NOS (iNOS) in the brain tissues were determined. The expression of neuronal NOS(nNOS) and iNOS was detected by Western blotting. RESULTS: Compared with sham-operation group, ginsenoside Rg1 significantly reduced the neurological deficit score and increased the neuron number in the hippocampus. The activity of NOS and iNOS, and NO content were decreased. Ginsenoside Rg1 also down-regulated the expression of nNOS and iNOS. CONCLUSION: Ginsenoside Rg1 has protective effect on the brain during cerebral I/R injury in rats. The mechanism may be related to reducing the content of NO and the activiy of NOS dose-dependently.     

Changes and significance of ATPase and free radical in aged rats with brain ischemia-reperfusion

AIM: To study the mechanism of brain ischemia-reperfusion injury from ATPase activity and free radical metabolism in aged rats. METHODS: The young rats (5 months) and the aged rats (more than 20 months) were divided into young control group(YCG), young model group(YMG), aged control group(ACG) and aged model group(AMG). The ATPase and SOD activities and the contents of MDA, Ca^2＋, Na^＋ and K^＋ were measured in the rats with 30 min brain ischemia followed by 60 min reperfusion. RESULTS: The Ca^2＋content in the AMG was higher than that in the YMG and the ACG. The Na^＋-K^＋-ATPase activity in the ACG was lower than that in the YCG,was lower in the AMG than that in the YMG. The Ca^2＋-ATPase activities in the YCG was higher than that in the ACG, was lower in the AMG than that in the YMG and was higher than the ACG's. The serum and brain tissue SOD activities in the ACG was lower than that in the YCG, was lower in the AMG than YMG 's. The serum and brain tissue MDA/SOD ratio in the AMG was higher than that in the ACG.CONCLUSION:The brain tissue ischemia-reperfusion injury was related with calcium overload and free radical injury.The pathological changes were obvious and had some characteristics in the aged rats compared with the young rats because of the brain t issue aging changes in ATPase,calcium content and free radical metabolism in the aged rats.     

Effect of cold water stress on memory and tau phosphorylation of rat

AIM: To explore the effect of cold water stress (CWS) on memory and tau phosphorylation in rat and their relationship. METHODS: Step-down test was used to measure the memory performance in rats at different time points after suffering from cold water stress. Levels of phosphorylated tau in different brain regions of rat subjected to CWS were detected by using immunoblotting. The relationship between memory and phosphorylated tau in rats was studied. RESULTS: The memory record of the rat obviously decreased from the 20 to 60 min after CWS, while trending to recover at the 90 and 120 min. At the same time, the levels of phosphorylated tau in different brain regions (hippocampus and cortex) in rat gradually increased at the 20, 40 and 60 min then was apt to recover at the 90 and 120 min. CONCLUSION: Cold water stress induces transient memory decrease in rat and corresponding increase in tau phosphorylation. It implies that tau phosphorylation may play a key role in memory formation in rat.     

Expression of synaptophysin in hippocampal CA1 region and prefrontal cortex of PTSD rats

AIM:To investigate the expression of synaptophysin in the CA1 region of hippocampus and prefrontal cortex (PFC) of rats with posttraumatic stress disorder (PTSD), and to explore the mechanism of spatial memory changes in PTSD rats.METHODS:Healthy adult SD rats (n=36) were randomly divided into 2 groups:control group and model group, with 18 rats in each group. The rats in model group was continuously given single prolonged stress (SPS) to construct the PTSD model. Morris water maze (MWM) was used to test the learning and memory ability of the rats in the 2 groups. The protein expression of synaptophysin in the hippocampal CA1 area and PFC was examined by immunohistochemistry, Western blot and immunofluorescence experiments. RESULTS:The latency of the rats in searching for the underwater platform in model group was significantly longer than that in control group from the 2nd day (P<0.01) in the MWM experiment, the target quadrant swimming time was significantly shortened (P<0.01), and the times of crossing the platform were also significantly reduced (P<0.01). The results of immunohistochemistry, Western blot and immunofluorescence experiments showed that the expression of synaptophysin was obviously reduced in the CA1 region of hippocampus and PFC in model group as compared with control group (P<0.05 or P<0.01).CONCLUSION:The reduction of spatial memory ability in PTSD rats may be associated with the decreased expression of synaptophysin in the CA1 region of hippocampus and PFC.     

Astrocytic connexin 43 is involved in development of cryoinjury-induced cerebral edema

ATM: To discuss the change of astrocytic connexin 43 (Cx43) expression in cerebral edema and the potential roles of Cx43 in the development of cerebral edema. METHODS: Cryoinjury-induced cerebral edema model was established in the right parietal lobe cortex in the rats. The animals were divided into sham group, cryoinjury-induced cerebral edema model group, and cryoinjury-induced cerebral edema model injected with carbenoxolone (or octanol) group. Dry and wet weight method was used to measure the content of brain water. Methanamide method was used to evaluate the permeability of blood-brain barrier (BBB) after cryoinjury. HE staining showed the physiological changes in injured brain cortex. The protein expression of Cx43 was detected by Western blot and immunohistochemical staining. RESULTS: The brain water content in the injured cortex of rats was increased and cerebral edema developed to the peak at 24 h after cryoinjury. The permeability of BBB was increased in the injured cortex of the rats, and the area where BBB was damaged was larger than injured area. HE staining showed that many cells were dead in the central area of cryoinjury, and at the same time cerebral edema was appeared obviously in the surrounding area of injured brain cortex. The protein expression of Cx43 was up-regulated in the surrounding area of injured brain cortex, while Cx43 expression decreased in the injured cortex at different periods after cryoinjury. Both the content of brain water in the injured cortex and the permeability of BBB at 24 h after cryoinjury were reduced after injecting carbenoxolone or octanol intraperitoneally to block Cx43. CONCLUSION: The cerebral edema up-regulates the expression of Cx43 and also strengthens its function. Once Cx43 is blocked, the degree of cerebral edema in rats is attenuated.     

Effect of chronic intermittent hypoxia on memory and CREB expression in growing rats

AIM: To observe the alterations in cognition of growing rats exposed to chronic intermittent hypoxia (CIH) and to explore its underlying mechanisms. METHODS: Forty male Sprague-Dawley rats (3-week-old～4-week-old and 80 g to 100 g), which had been trained to complete the 8-arm (4-arm baited) radial maze, were randomly divided into 4 groups: 2-weeek-CIH group (2IH), 4-week-CIH group (4IH), 2-week-control group (2C) and 4-weeek-control group (4C). The intermittent hypoxia model was induced by putting the animals in an intermittent hypoxia cabin. When intermittent hypoxia was terminated, spatial memory of these growing rats was tested by 8-arm (4-arm baited) radial maze task, then, one rat in each group was randomly selected for ultrastructural observation. The hippocampus and prefrontal cortexes of the rats were collected for analyzing the mRNA and protein expression of CREB by RT-PCR and Western blotting, respectively. RESULTS: (1) In the 8-arm (4-arm baited) radial maze task, the results indicated that the rats in the 4 groups displayed significant difference in their performance assessed by three measuremens: the reference memory error, the working memory error and total memory error (P<0.05, respectively). (2) Early apoptosis and destructure of the neurons in the hippocampus and prefrontal cortex were observed under electron microscope in CIH exposed groups, especially in 4IH group, but not detected in 2C and 4C groups. (3) The expression levels of CREB mRNA and p-CREB protein in 2IH and 4IH groups were less than those in 2C and 4C groups in the hippocampus and prefrontal cortex (P<0.05, respectively), especially in the hippocampus of 4IH group (P<0.01). No difference was found within control groups (P>0.05, respectively). CONCLUSION: Exposure to experimentally-induced IH in growing rats is associated with time related spatial memory impairment. Chronic intermittent hypoxia leads to the disorders of neuron ultra-structure in memory related brain regions. It also inhabits the CREB transduction, expression and CREB phosphorylation, decreases the synthesis of the memory related protein. These factors maybe contribute to learning-memory impairment of growing rats exposed to chronic intermittent hypoxia.     

Early-onset type 1 diabetes causes brain injury and mitochondrial dysfunction

AIM:To explore the main risk factors of diabetic encephalopathy and to investigate the changes of mitochondria in early-onset type 1 diabetes. METHODS:Single dose of streptozotocin was injected through the femoral vein to establish type 1 diabetes model in rats. The levels of glucose, triglyceride and cholesterol in plasma, cerebrospinal fluid and cerebral cortex were measured. Oxidative and antioxidative status was evaluated by determining the levels of malondialdehyde and glutathione in the isolated mitochondria of cerebral cortex. Furthermore, the level of active AMP-activated protein kinase(AMPK) in the isolated mitochondria was detected by Western blotting. Degenerative neurons were identified by Fluo-Jade-C staining in serial brain sections. Autophagy-lysosome was observed under electron microscope. RESULTS:The main risk factor in the development of diabetic encephalopathy was hyperglucemia, which increased the Fluo-Jade-C positive neurons in the cerebral cortex of diabetic rats. The content of malondialdehyde was increased and glutathione was decreased in diabetic rats compared with the control animals. The activity of AMPK was lower in diabetic brain than that in normal brain. Aggregated autophagysome and mitochondria enveloped by autophagy-lysosome were observed in pyramidal cells of cerebral cortex in diabetic rats. CONCLUSION:Persistent hyperglycemia and mitochondrial dysfunction contribute to the diabetic encephalopathy at an early stage in type 1 diabetes, indicating that the mechanism may be partly related to the oxidative stress and activation of autophagy.