Effects of high copper diet on cognitive function and hippocampal synaptic protein in rats / 中华行为医学与脑科学杂志

Objective:To observe the effects of high copper diet on neurobehavioral functions and synaptic associated protein expression in hippocampus of rats.Methods:Thirty male SD rats were randomly divided into control group and high copper diet group with 15 rats in each group according to the random number table method. The rats in control group were fed with ordinary diet and ordinary water, while the rats in high-copper diet group were fed with high-copper diet containing 1 g/kg copper sulfate and 0.185% copper sulfate deionized water for 12 weeks. The content of copper in serum and hippocampus of rats were detected by inductively coupled plasma-atomic emission spectrometry(ICP-AES) and ICP-mass spectrometry(ICP-MS). The neurobehavioral indicators were detected by stereotypic behavior test, open field test and Morris water maze test. The expression levels of microtubule associated protein 2(MAP2) and growth associated protein 43 (GAP43) in hippocampus were detected by Western blot.SPSS 22.0 software was used for statistical analysis, and two independent sample t-test was used for comparison between the two groups. Results:Compared with the control group, the content of serum copper((1.67±0.69)mg/L, (1.98±0.24)mg/L, t=17.53, P<0.05) and hippocampal free copper((3.52±1.24)mg/g, (4.78±0.57)mg/g, t=10.34, P<0.05) in the high copper diet group increased significantly, and the stereotypic behavior score increased significantly ((0.29±0.08), (2.97±0.72), t=14.33, P<0.01), the number of space crossing in the open field experiment ((153.40±24.73)points, (92.46±19.46)points, t=7.50, P<0.01) and the times of standing((19.34±1.98)times, (10.57±2.71)times, t=10.12, P<0.01) were significantly decreased. The average latency in Morris water maze navigation test was significantly prolonged ((3.14±1.67)s, (8.29±2.26)s, t=7.10, P<0.01), the number of crossing the original platform position in the space exploration test decreased significantly ((7.89±2.48)times, (2.98±1.73) times, t=3.23, P<0.01). Compared with control group, protein levels of GAP43((1.03±0.05), (0.48±0.02), t=39.56, P<0.05)and MAP2((0.93±0.05), (0.30±0.08), t=25.86, P<0.05) of high copper diet group were significantly decreased. Conclusion:High copper diet causes abnormality in a variety of neurobehavioral function indexes in rats, and a decrease in expression of MAP2 and GAP43 at the synaptic interface of hippocampal neurons may be involved in the process of learning and memory impairment in the neurobehavioral functions.

Repair effect of wogonoside on rat spinal cord injury / 解剖学报

[Abstract] Objective To assess the effect of wogonoside on the inflammation after rat spinal cord injury. Methods Rats (n = 95) were subjected to dorsal spinal cord transection at T

Effect of moxibustion on expression of GAP-43 in sciatic nerve trunk and ventral horn of spinal cord of rats with primary sciatica and its nerve repair function / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion at "Huantiao" (GB 30) on the expression of growth-associated protein-43 (GAP-43) in the sciatic nerve trunk and ventral horn of spinal cord (L@*METHODS@#A total of 48 healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and a moxibustion group, 12 rats in each group. The rat model of primary sciatic pain was established by chronic constriction injury (CCI) of the sciatic nerve in the model group and the moxibustion group. On the 8th day of the experiment, moxibustion was adopted at "Huantiao" (GB 30) in the moxibustion group for 5-10 min, once a day for 14 consecutive days. Sciatic nerve function index (SFI) was measured and compared in each group at day 1, 7, 14 and 21. On the 21st day of the experiment, HE staining was used to observe the morphology of ventral horn of rat spinal cord and sciatic nerve trunk. Immunohistochemical method and real-time PCR were used to detect mRNA and protein expressions of GAP-43 in the spinal cord and sciatic nerve trunk of rats.@*RESULTS@#On day 7, 14 and 21, there was no statistical difference in SFI between the sham operation group and the normal group (@*CONCLUSION@#Moxibustion at "Huantiao" (GB 30) could improve the sciatic nerve function in rats with primary sciatica and its mechanism may be related to improving the expression of GAP-43 and enhancing the self-repair ability of the sciatic nerve after injury.

Effects of Ganoderma Triterpenoids on Learning and Memory Impairment in Rats with Epilepsy / 中国康复理论与实践

Objective:To study the effect of Ganoderma triterpenoids combined with exogenous monosialoteterahexosyl ganglioside (GM1) on cognitive dysfunction in rats with epilepsy. Methods:A total of 75 Sprague-Dawley rats were divided randomly into blank control group, epileptic model group, Ganoderma triterpenoids group, GM1 group and GM1 combined with Ganoderma triterpenoids group (combination group), with 15 rats in each group. All the groups, except the blank control group, were intraperitoneally injected with pentylenetetrazol (PTZ) 35 mg/kg once a day for 28 days. Medication groups were given corresponding administration based on daily intraperitoneal injection of PTZ. They were tested with Morris Water Maze; and were observed with transmission electron microscopy and HE staining for hippocampal neurons. Real-time quantitative polymerase chain reaction was used to detect the expression of actin-binding protein (Cofilin), synaptophysin (SYN) and growth-associated protein 43 (GAP-43) mRNA in hippocampus of rats. Results:Compared with the blank control group, the escape lantency prolonged in the epileptic model group in all the time points (P < 0.05). Compared with the epileptic model group, the escape lantency shortened in the treatment groups somewhen (P < 0.05). Compared with the epileptic model group, the number of crossing the platform increased in the treatment groups (P < 0.01), and the time of staying in the target quadrant prolonged (P < 0.01); while the number of pyramidal cells increased, the nuclear lysis and fragmentation reduced, the structure of neurons and the number of synapses improved， as well as the organelle structure. Compared with the blank control group, the expression of Cofilin mRNA increased (P < 0.05), and the expression of SYN mRNA and GAP-43 mRNA decreased (P < 0.05) in the epileptic model group; compared with the epileptic model group, the expression of Cofilin mRNA decreased (P < 0.05), and the expression of SYN mRNA increased (P < 0.05) in all the treatment groups, while the expression of GAP-43 mRNA increased (P < 0.05) only in the combination group. Conclusion:Ganoderma triterpenoids, GM1 and their combination can improve the learning and memory abilities of epileptic rats, which may be associated with increasing the expression of SYN and GAP-43, decreasing the expression of Cofilin, to promote the synaptic remodeling of hippocampal tissue and protect brain neurons from PTZ-induced epilepsy.

Effect of Buyang Huanwu Tang on Synaptic Structural Plasticity After Cerebral Ischemia-reperfusion in Rats / 中国实验方剂学杂志

<b>Objective::To investigate the mechanism of Buyang Huanwu Tang (BYHWT) in improving synaptic structural plasticity after cerebral ischemia-reperfusion in rats. <b>Method::Middle cerebral artery occlusion and reperfusion model was established. SD rats were randomly divided into sham-operated group, model group, BYHWT group, BYHWT+ Gap26(connexin43 inhibitor)groups. BYHWT was given twice a day(16 g·kg^-1), Gap26 was intraperitoneally injected once a day since the third day after surgery (25 g·kg^-1). Brain was taken out at the 7^th day. The changes of neuronal synaptic and gap junction ultrastructure were observed by transmission electron microscopy. Synaptophysin (SYN) and growth-associated protein-43 (GAP-43) protein expression were detected by Western blot and immunofluorescence. <b>Result::The structure of synapses was integrated, and the gap junctions were clear in sham-operated group. In the hippocampus of model group, the structure was destroyed, and the gap junctions disappeared. Compared with the sham-operated group, model group up-regulated the expressions of SYN and GAP-43 (<italic>P</italic><0.05, <italic>P</italic><0.01). In the hippocampus of BYHWT group, the structure was close to the normal. Furthermore, BYHWT up-regulated the expressions of SYN and GAP-43 (<italic>P</italic><0.05, <italic>P</italic><0.01). However, after the combined administration with Cx43 inhibitor (Gap26), the damage of synaptic structural decreased, only a small number of gap junctions with the structural integrity can be seen, and the effect of BYHWT on SYN and GAP-43 was inhibited (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::BYHWT could improve the hippocampal synaptic structural plasticity obviously after the CIRI. The mechanism may be related to the increase of the expression of Cx43 and the promotion of the intervention of SYN and GAP-43.

Effects of electroacupuncture on sympathetic nerve-related substance in myocardial tissue in mice with myocardial ischemia / 中国针灸

OBJECTIVE@#To observe the effects of electroacupuncture (EA) on sympathetic nerve-related substance in myocardial tissue in mice with myocardial ischemia (MI), and to explore its possible mechanism.@*METHODS@#Thirty adult male C57BL/6 mice were randomly divided into a sham operation group, a model group and an EA group, 10 mice in each one. The model of MI was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the sham operation group were not treated with ligating at left anterior descending branch of coronary artery, but the remaining procedure was similar with the model group. The mice in the EA group were treated with EA at "Neiguan" (PC 6) with 2 Hz/100 Hz of frequency and 2 mA of intensity, 20 min per treatment, once a day for totally 5 days. No EA was given for model group and sham operation group. The electrocardiogram was recorded and △ST value was calculated to evaluate the model. TTC staining was applied to evaluate the infarct size. Immunohistochemical (IHC) method was applied to evaluate the positive nerve fiber density in myocardial tissue. Western blot method was applied to test the protein expression levels of neuregulin-1 (NRG-1), tyrosine hydroxylase (TH), growth associated protein-43 (GAP-43).@*RESULTS@#The electrocardiogram (lead II) results indicated compared with the sham operation group, the S-T segments in the model group and EA group were increased obviously (both <0.01), indicating the MI model was established successfully. The TTC staining results indicated compared with sham operation group, the infarction size was significantly increased in the model group (<0.01); compared with the model group, the infarction size in the EA group was significantly reduced (<0.01). The IHC results indicated compared with the sham operation group, the positive nerve fiber density in myocardial was increased in the model group (<0.01); compared with the model group, the positive nerve fiber density in myocardial was reduced in the EA group (<0.05). The Western blot results indicated compared with the sham operation group, the expression levels of TH, NRG-1 and GAP-43 were significantly increased in the model group (<0.01); compared with the model group, the expression level of TH and GAP-43 were significantly reduced (<0.01) and that of NRG-1 was increased in the EA group (<0.05).@*CONCLUSION@#EA could increase the expression of NRG-1 and reduce the expression of TH and GAP-43 in myocardial tissues in MI mice, which could suppress sympathetic nerve hyperexcitability after infarction to achieve myocardial protection effect.

Effect of botulinum neurotoxin type A heavy chain on the growth-related proteins after spinal cord injury in rats / 中国比较医学杂志

Objective To investigate the effect of recombinant botulinum neurotoxin serotype A heavy chain (BoNT/A heavy chain)on local proteins which are related to nerve growth after spinal cord injury in rats,and to get some experimental evidence to explain the mechanism of BoNT/A heavy chain in stimulating neuritogenesis. Methods Recombinant botulinum neurotoxin serotype A heavy chain was applied locally or intrathecally to rats with ipsilateral semi-dissociated lumbar spinal injury. Local spinal tissue was extracted for general protein expression by two dimension electrophoresis plus nitrate silver staining after different time period of injury. Based on the results of 2-D gel electrophoresis,growth-associated protein 43(GAP-43)and of superior cervical ganglion 10(SCG 10)were selected to examine the changes of their expression and distribution features under BoNT/A heavy chain administration using SDS-PAGE,western blot and immunofluorescence. Results (1)The model of spinal cord injury(SCI)in this study was an ipsilateral semi-dissociated lumbar SCI in rat. The rats showed obvious motor and sensory dysfunction in the ipsilateral hind limb.(2)The results from 2-D gel electrophoresis plus nitrate silver staining showed that the administration of BoNT/A heavy chain based on SCI altered the local protein expression pattern. The decrease or increase in the expression of some protein dots /dots group was clearly seen after single SCI. However, these changes were transformed by BoNT/A heavy chain treatment,which appeared as a reversed pattern turning toward that in control group or further increased expression upon SCI,such as the dots located respectively at 35-45 kDa and 18-25 kDa level,pI between 5-7. In addition,the expression of the two dots located at the level as above increased after SCI only, and showed further increase in their expression with BoNT/A heavy chain intervention.(3)The changes of selective GAP-43 and SCG 10 expression and distribution by western blot and immunofluorescence indicated that the administration of BONT/A heavy chain based on SCI amplified the expression of GAP-43 and SCG 10(P < 0.05). Meanwhile,the positive immuonfluorescent staining for both GAP-43 and SCG 10 mainly distributed nearby the proximal area of injury, both cytoplasm and neuronal processes were positively stained. Conclusions Intrathecal delivery of BoNT/A heavy chain increases the expression of growth-associated proteins GAP 43 and SCG 10 after SCI in rats.

The effect of electroacupuncture combined with repetitive transcranial magnetic stimulation on rats' learning and memory after middle cerebral artery occlusion and reperfusion / 中华物理医学与康复杂志

Objective To study the effect of electroacupuncture combined with repetitive transcranial magnetic stimulation on the infarct volume of rats with cerebral infarction as well as their behavior and their learning and memory ability.Methods Forty male Sprague-Dawley rats were randomly divided into a normal group,a sham-operation group,a model group and a treatment group,each of 10.Focal cerebral ischemia and reperfusion was modeled in the latter two groups using thread occlusion of the middle cerebral artery.The treatment group was treated for 14 days with electroacupuncture combined with repetitive transcranial magnetic stimulation,started 24 hours after the operation.The animals' neurological functioning was quantified using modified neurological severity scores.The area of cerebral infarct was measured using 5-triphenyltetrazolium chloride staining.The rats' learning and memory abilities were assessed using a Morris water maze and the expression of growth-associated protein 43 (GAP-43) in the peripheral area of the cerebral infarction.Results The average neurological deficit score (2.8± 0.84) was significantly lower in the treatment group than in the model group,the average infarct volume was smaller and the escape latency was less.In the treatment group the average expression of GAP-43 was greater than in the model group.Conclusion Electroacupuncture combined with repetitive transcranial magnetic stimulation can promote the expression of GAP-43 in the peripheral area of a cerebral infarct,promoting neuronal remodeling and thereby improving neurological functioning as well as learning and memory ability.

The effect of Puerarin on GAP-43 protein and NGF protein expression of spinal motoneurons following brachial roots avulsion / 中华显微外科杂志

Objective To study the effects of Puerarin(Pue)on survival rate,expression of GAP-43 and NGF in spinal motoneurons following brachial roots avulsion. Methods From March, 2014 to December, 2015, 192 adult Sprague Dawley rats were divided into four groups: Avulsion, Pue 50 treatment, Pue 100 and Pue 200 groups. The right C5-C7nerve roots were avulsed through the methods of cervical dorsal approach. Puerarin or normal saline was given immediatedly once daily to the rats respectively by the intraperitoneal injection. The rats were killed at 1, 2, 4 and 6 weeks after injury. The paraffin sections of C7 segment were stained with neutral red. Expression of GAP-43 and NGF were detected by Western blot. Results Compared to the avuled group, the motoneuron survival rates of Pue 100 and Pue 200 dose treatment groups increased on the second week and the sixth week(P < 0.05 or P < 0.01), and the Pue 200 dose treatment group increased on the fourth week(P < 0.01).In the anterior horn of all three groups, expression of GAP-43 increased from the 1st week to the 6 week after the operation, and reached the peak at the 4th week, and decreased at the 6th week. Compared to the avuled group, expression of GAP-43 protein increased in the Pue 100 and Pue 200 dose treatment groups at the 1st week and 2nd week,the expression of NGF protein increased in the three Pue treatment groups at four time points(P < 0.05 or P < 0.01). Conclusion Puerarin can improve the survival rates of spinal motoneurons after the brachial plexus root avulsion, and the expression of GAP-43 and NGF increased, suggesting that Puerarin may play an role in the repair of brachial plexus avulsions by promoting the ex-pression of nerve growth factors.

Analgesic effect of angiotensin angiotensin Ⅱ type 2 receptor antagonist EMA401 on neuropathic pain in rats and its mechanism / 中国病理生理杂志

AIM:To explore whether angiotensin Ⅱtype 2 receptor antagonist EMA 401 decreases neuropathic pain and the expression of growth-associated protein-43 (GAP-43), protein kinase C (PKC) and calmodulin (CaM) in dorsal root ganglia (DRG) during chronic constriction injury (CCI) in rats.METHODS:SD rats were used to establish CCI model and randomly divided into 4 groups.The rats in model group were given equal volume of normal saline by intra-gastric administration .The rats in low dose ( LD) group were given 5 mg/kg EMA401 by intragastric administration .The rats in middle dose ( MD) group were given 10 mg/kg EMA401 by intragastric administration .The rats high dose ( HD) group were given 20 mg/kg EMA401 by intragastric administration .The rats in sham operation group received equal volume of normal saline by intragastric administration .Thermal withdrawal latency ( TWL ) and mechanical withdrawal threshold (MWT) were measured before operation and 7 d, 14 d and 28 d after CCI.After behavioral test, DRG of lumbar spinal was obtained from each group , and was used to determine Ca 2+concentration by o-cresolphthalein complexone microplating method, and the expression of GAP-43, PKC and CaM at mRNA and protein levels by Western blotting and RT-PCR.RE-SULTS:Compared with model group, EMA401 significantly increased the TWL and MWT (P <0.05).Meanwhile, EMA401 significantly reduced Ca 2+concentration and the expression of GAP-43, PKC and CaM at mRNA and protein levels in the DRG (P<0.05).CONCLUSION:EMA401 may attenuate neuropathic pain of CCI by inhibiting Ca 2+concentra-tion and the expression of GAP-43, PKC and CaM.

Effects of curcumin on the cellular model of Alzheimer′s disease and GAP-43 expression / 国际药学研究杂志

Objective To investigate the protective effects of curcumin on the cellular model of Alzheimer's disease(AD) and the expression of growth associated protein-43(GAP-43). Methods Aβ25~35 was used to treat the hippocampus neurons of rat and the cellular model of AD was established. The survival rate was detected by MTT assay. The cells were randomly divided into blank control,model,curcumin 10 and 20μmol/L groups. The effect of curcumin on apoptosis was assayed by flow cytometry. The pro?trusive length and GAP-43 positive cell rate were detected by immunocytochemistry. The expression of GAP-43 was detected by West?ern blot. Results Compared with the model group,curcumin significantly reduced the toxicity of Aβ25~35,increased the survival rate and decreased the apoptosis rate of the cells(P<0.05). It also significantly increased the average protrusive length,GAP-43 positive cell rate and GAP-43 expression(P<0.05 or P<0.01). Conclusion The protective effect of curcumin on the cellular model of AD was likely related to the up-regulation of GAP-43 expression.

Effects and mechanisms of different frequencies of electroacupuncture for learning and memory ability of Alzheimer's rats / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of different frequencies of electroacupuncture (EA) at "Baihui (GV 20)" and "Shenshu(BL 23)" for the learning and memory ability as well as glycogen synthase kinase-3β (GSK-3β) and growth associated protein-43 (GAP-43) in hippocampal tissue of rats with Alzheimer's disease(AD), so as to explore the mechanism of different frequencies of EA for the prevention and treatment of AD.</p><p><b>METHODS</b>One hundred and twelve healthy Wistar male rats were divided into seven groups by random number table, namely a normal group, a sham operation group, a model group, an acupuncture group, a 2 Hz EA group, a 30 Hz EA group, and a 50 Hz EA group, 16 rats in each one. The rats in the normal group were conventionally raised in the laboratory without any treatment. 0.9% NaCl solution was injected into bilateral dentate convolution of hippocampus in rats of the sham operation group. AD model was established by β-amyloid protein1-42 (Aβ1-42) injected into bilateral dentate convolution of hippocampus in the other groups. 15 days after establishment, no treatment was applied in the model and sham operation groups, and EA with corresponding frequencies at "Baihui (GV 20)" and "Shenshu (BL 23)" was used in the three EA groups for 2 sessions, once a day and 7 times as one session. There was 1 day between the two sessions. The same acupoints were adopted in the acupuncture group, without electrical connection. The escape latency, the first spanning platform time, and the number of crossing platform were tested in the Morris water maze immediately after treatment. The expressions of GSK-3β and GAP-43 were examined by immunohistochemistry and Western blot.</p><p><b>RESULTS</b>①Morris water maze tests showed that the escape latency and the first spanning platform time significantly increased in the model group compared with those in the normal group (both<0.01), and the number of crossing platform decreased (<0.01). Compared with the model group, the escape latency and the first spanning platform times decreased in the acupuncture and three EA groups (all<0.01), and the numbers of crossing platform increased (<0.01). Compared with the acupuncture and 2 Hz, 30 Hz EA groups, the escape latency decreased in the 50 Hz EA group (<0.01,<0.05); the first spanning platform time reduced (all<0.01); the number of crossing platform increased (<0.01,<0.05). ②The expressions of GSK-3β and GAP-43 of the model group increased compared with those of the normal group(both<0.01). The expressions of GSK-3β in the acupuncture and three EA groups decreased compared with that in the model group (all<0.01), and the expressions of GAP-43 increased (all<0.01). The expressions of GSK-3β decreased and GAP-43 increases in the 50 Hz EA group compared with those in the acupuncture group and 2 Hz, 30 Hz groups (all<0.01).</p><p><b>CONCLUSIONS</b>EA may promote synaptic damage rehabilitation by down regulating GSK-3β and up regulating GAP-43 to improve learning and memory ability of AD rats. The effect of 50 Hz EA is better than those of 30 Hz and 2 Hz EA and acupuncture.</p>

Effects of curcumin on the cellular model of Alzheimer′s disease and GAP-43 expression / 国际药学研究杂志

Objective To investigate the protective effects of curcumin on the cellular model of Alzheimer’s disease(AD) and the expression of growth associated protein- 43(GAP- 43). Methods Aβ2535 was used to treat the hippocampus neurons of rat and the cellular model of AD was established. The survival rate was detected by MTT assay. The cells were randomly divided into blank control, model, curcumin 10 and 20 μmol/L groups. The effect of curcumin on apoptosis was assayed by flow cytometry. The protrusive length and GAP-43 positive cell rate were detected by immunocytochemistry. The expression of GAP-43 was detected by Western blot. Results Compared with the model group, curcumin significantly reduced the toxicity of Aβ25~35, increased the survival rate and decreased the apoptosis rate of the cells(P<0.05). It also significantly increased the average protrusive length, GAP-43 positive cell rate and GAP-43 expression(P<0.05 or P<0.01). Conclusion The protective effect of curcumin on the cellular model of AD was likely related to the up-regulation of GAP-43 expression.

Expression of Growth Associated Protein 43 in Inferior Colliculus of Rats at Different Times after Impulse Noise Exposur / 听力学及言语疾病杂志

Objective To investigate the effect of impulse noise expose on the expression of growth associated protein 43(Gap-43) in inferior colliculus in rat.Methods SPF grade Male SD rats were randomly divided into 5 groups.The normal control group received noise exposure.The model groups received an averange impulse noise exposure of 156 dB SPL with a pulse duration of 0.23 ms, once for 6 s, for 50 times.Auditory brainstem responses (ABRs) were measured before and 3,7,14, and 28 d after noise exposure with tone pips of 2,4,8,16, and 32 kHz, from 20 to 110 dB SPL.Bilateral inferior colliculus of rats in the model groups was collected and treated by immunohistochemical staining.Gap-43 expression of rats in different groups was measured by determining the gray value of inferior immunohistochemical images.Results After noise exposure, ABRs threshold in the model groups were significantly higher than those of in the normal group (P<0.05 or P<0.01).ABRs threshold at 14 and 28 days after noise exposure were significantly lower than 3 days after impulse exposure (P<0.05).Expression of Gap-43 in inferior colliculus was significantly up-regulated in the noise exposed groups compared with the normal group (P<0.05 or P<0.01).Expression of Gap-43 was significantly down-regulated 28 days after noise expose compared with 3 days after noise expose(P<0.05).Conclusion Impulse noise exposure leads to significant elevation of ABR thresholds and up-regulation of Gap-43 expression in inferior colliculus.Impulse noise exposure may induce auditory cortex prominent remodeling.

Carbamylated erythropoietin can promote neural regeneration after cerebral ischemia / 中华物理医学与康复杂志

Objective To investigate any effects of carbamylated erythropoietin (CEPO) on the expression of LINGO-1,growth-associated protein-43 (GAP-43) and the infarcted volume after cerebral ischemia,so as to explore the effect of CEPO on neural regeneration after cerebral ischemia.Methods Forty-eight Sprague-Dawley rats were randomly divided into a sham operation group,an ischemia control group and a CEPO treatment group,each of 16.Middle cerebral artery occlusion (MCAO) was used to simulate focal cerebral ischemia in all except the rats in the sham operation group.Then the CEPO group was injected with 0.5 ml of CEPO,while the other two groups were given a 0.5 ml injection of normal saline daily for 7 days before they were sacrificed to prepare slices of brain tissue.Western blotting was used to detect the expression of LINGO-1 and activated caspase-3.Immunohistochemical staining was applied to observe the expression of GAP-43.The slices of brain tissue were stained with cresyl violet and the volume of infarction and edema were quantified with the Image J software.Results The average expression of LINGO1 in the sham operation group,the ischemia control group and the CEPO treatment group were (0.25±0.02),(1.22±0.06) and (0.66±0.05) respectively,with significant differences among the 3 groups.There was no expression of activated caspase-3 in the sham operation group.However,the expression of activated caspase-3 increased significantly (to 86.6±10.2)％ in the ischemia control group and increased significantly less (to 40.3±8.7)％ in the CEPO treatment group.The average positive expression of GAP-43 in the sham operation group,the ischemia control group and the CEPO treatment group were 0,(55.02± 1.62) and (72.11±3.23)/HP,respectively,with significant differences among them.Moreover,the average volumes of cerebral infarction and brain edema in the CEPO treatment group were significantly lower than those in the ischemia control group.Conclusions CEPO can inhibit the expression of LINGO-1 and activated caspase-3,promote the expression of GAP-43,reduce infarct volume and limit cerebral edema so as to promote neural regeneration after cerebral ischemia,at least in rats.

Effect of glucocorticoid on the retinal neurons of diabetes mellitus rats / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the detrimental effect of glucocorticoid(GC)on the retinal neurons of diabetes mellitus(DM)rats.<p>METHODS: The DM model was induced by intraperitoneal injection(IP)of streptozotocin in adult male rats, and the solution of RU486 was configured with dimethyl sulfoxide(DMSO). RU486 treatment group with glucocorticoid receptor antagonist RU486 and diabetic group with DMSO by intraperitoneal injection was in successful DM model. Naive rats were injected with DMSO as control group. Three months later, we detected the body weight and blood glucose and GC concentration of serum. The changes of retinal ganglion cell(RGC)density was investigated by HE staining. The expression of growth associated protein-43(GAP-43, a marker of neuronal axon regeneration)and synaptophysin(SYN, a marker of synaptic number)were semi-quantity analyzed by the optical density of immunofluorescence and Western blot.<p>RESULTS:Compared with the control group, the body weight and density of RGC and expression of SYN in diabetic group were significantly lower(<i>P</i><0.01), the blood glucose and GC concentration of serum and expression of GAP-43 in diabetic group were significantly higher(<i>P</i><0.01). Compared with the diabetic group, the density of RGC and expression of GAP-43, SYN in RU486 group were significantly higher(<i>P</i><0.01).<p>CONCLUSION: Inhibition of GC could ameliorate the axonal degeneration of retinal neurons in diabetic rats, and loss of the number of synapses, and restore the RGC density of retina. The results suggest that long-term elevation of GC may be involved in the occurrence and development of diabetic retinopathy.

Hippocampus-dependent Task Improves the Cognitive Function after Ovariectomy in Rats

OBJECTIVES: Estrogen is an important hormone for cell growth, development, and differentiation by transcriptional regulation and modulation of intracellular signaling via second messengers. The reduction in the estrogen level after ovariectomy may lead to cognitive impairments associated with morphological changes in areas of the brain mediate memory. The aim of the present study was to find out the effect of tasks on the cognitive function after ovariectomy in rats. METHODS: The animals used in the experiment were 50 Sprague-Dawley female rats. This study applied a hippocampus-independent task (wheel running) and a hippocampus-dependent task (Morris water maze) after ovariectomy in rats and measured the cognitive performance (object-recognition and object-location test) and growth-associated protein 43 (GAP-43) and neurotrophin 3 (NT-3) expression in the hippocampus, which is an important center for memory and learning. RESULTS: There were meaningful differences between the hippocampus-independent and hippocampus-dependent task groups for the object-location test and GAP-43 and NT-3 expression in the hippocampus, but not the object-recognition test. However, the hippocampus-independent task group showed a significant improvement in the object-recognition test, compared to the control group. CONCLUSION: These results suggest that hippocampus-dependent task training after ovariectomy enhances the hippocampus-related memory and cognitive function that are associated with morphological and functional changes in the cells of the hippocampus.

Effect of Aerobic Exercise Preconditioning on Growth-associated Protein-43 and Nogo-A in Rats after Cerebral Ischemia/Reperfusion / 中国康复理论与实践

Objective To observe the effect of aerobic exercise preconditioning on growth-associated protein-43 (GAP-43) and Nogo-A in rats after cerebral ischemia/reperfusion. Methods Sprague-Dawley rats were equally divided into sham group (n=40), cerebral ischemia/reperfusion group (n=40) and aerobic exercise preconditioning group (n=40), and global cerebral ischemia model was formed with modified four-vessel occlusion. The rats was sacrificed six hours, one day, three days and seven days after ischemia, respectively. The hippocampus neural cells were observed in five rats with HE staining and immunohistochemistry of GAP-43 and Nogo-A, and the other five rats were test-ed with RT-PCR of GAP-43 and Nogo-A. Results Compared with those in the cerebral ischemia/reperfusion group, the apoptotic neurons and expression of GAP-43 significantly increased all the time points in the aerobic exercise preconditioning group (P<0.01), while the ex-pression of Nogo-A decreased (P<0.01). Conclusion Aerobic exercise preconditioning can promote the regeneration of neuronal cells and axon after cerebral ischemia/reperfusion injury, which is related to the regulation of GAP-43 and Nogo-A.

Effects of Autophagy on Expression of Growth-associated Protein-43 and Microtubule Associated Protein-2 in CA1 Area of Hippocampus of Vascular Dementia Rats / 中国康复理论与实践

Objective To observe the effects of autophagy on the expression of synaptic plasticity related protein, growth-associated pro-tein-43 (GAP-43) and microtubule associated protein-2 (MAP-2), in CA1 area of hippocampus of vascular dementia rats. Methods Nine-ty-six healthy male Sprague-Dawley rats were randomly divided into sham group, vascular dementia model group (VD group), autophagy in-hibitor 3-methyl adenine preconditioning group (3-MA group) and autophagy agonist rapamycin preconditioning group (Rap group). Each group was divided randomly into subgroups of one week, two weeks, four weeks and eight weeks after modeling, six rats in each group. The vascular dementia rat model was established with modified Pulsineli's four-vessel occlusion. The expression of GAP-43 and MAP-2 in CA1 area of hippocampus were detected with immunohistochemistry. Results Compared with the sham group, the expression of GAP-43 protein increased, and the expression of MAP-2 protein decreased at every time point in VD group (P<0.01). Compared with VD group, the expres-sion of both GAP-43 and MAP-2 increased in 3-MA group (P<0.05), and decreased in Rap group (P<0.05). Conclusion Autophagy may in-hibit the expression of synaptic plasticity related protein, GAP-43 and MAP-2, in CA1 area of hippocampus in vascular dementia rats, indi-cating inhibition of autophagy may promote synaptic remodeling.

Protective effect of Grape seed proanthocyanidin on learning disabilities in a rat model with radiation brain injury / 中华行为医学与脑科学杂志

Objective To observe the protective effect of grape seed proanthocyanidin extract ( GSPE) on learning disabilities after radiation injury brain in rats.Methods 120 male Wistar rats were divided into control group,model group,high and low dose GSPE groups.The radiation injury brain models were established using a method of linear accelerator irradiation in 22 Gy.The morphological changes of neu-rons in hippocampus were observed with HE staining; the expression of phosphorylated extracellular signal regulated kinase 1/2 ( ERK1/2) and growth associated protein-43( GAP-43) were detected by immunohisto-chemistry;the contents of malonalde hyde(MDA) and superoxide dismutase(SOD) were measured respec-tively by thiobarbituric acid( TBA) and xanthine oxidation( XTO) methods;the learning ability was assessed with shuttle box.Results Compared with the model group,the GSPE groups showed a decreased degree of nerve cell morphological injury; and increased ERK1/2 activities((13.20±1.45)/view,(27.40±2.52)/view,(19.80±1.30)/view),GAP-43 expression level((52.93±2.07)/view,(66.50±0.77)/view,(73.05± 2.40)/view) and SOD activities((79.82±5.26) U/mg,(76.20±6.86) U/mg,(80.12±5.76) U/mg)(P<0.05);and decreased the contents of MDA((71.62±1.88) μmol/g,(76.41±1.94) μmol/g,(72.32±1.98)μmol/g)(P<0.05).Shuttle box testing showed that the active avoidance reaction rate was increased(7 d:(56.23±2.56)%;14 d:(44.66±2.79)%;28 d:(50.40±2.16)%)(P<0.05)and passive avoidance latency was shorted(7 d:(34.11±1.57)s;14 d:(45.52±1.88)s;28 d:( 39.52±1.79)s)(P<0.05) in GSPE groups. The above mentioned changes were more significant in high dose of GSPE(P<0.05) .Conclusion GSPE has protective effect on learning disabilities of radiation injuries brain,which is related to enhance ERK1/2 activ-ity and the expression of GAP-43.