Protective effect of Epothilone D against traumatic optic nerve injury in rats / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic effect of Epothilone D on traumatic optic neuropathy (TON) in rats.@*METHODS@#Forty-two SD rats were randomized to receive intraperitoneal injection of 1.0 mg/kg Epothilone D or DMSO (control) every 3 days until day 28, and rat models of TON were established on the second day after the first administration. On days 3, 7, and 28, examination of flash visual evoked potentials (FVEP), immunofluorescence staining and Western blotting were performed to examine the visual pathway features, number of retinal ganglion cells (RGCs), GAP43 expression level in damaged axons, and changes of Tau and pTau-396/404 in the retina and optic nerve.@*RESULTS@#In Epothilone D treatment group, RGC loss rate was significantly decreased by 19.12% (P=0.032) on day 3 and by 22.67% (P=0.042) on day 28 as compared with the rats in the control group, but FVEP examination failed to show physiological improvement in the visual pathway on day 28 in terms of the relative latency of N2 wave (P=0.236) and relative amplitude attenuation of P2-N2 wave (P=0.441). The total Tau content in the retina of the treatment group was significantly increased compared with that in the control group on day 3 (P < 0.001), showing a consistent change with ptau-396/404 level. In the optic nerve axons, the total Tau level in the treatment group was significantly lower than that in the control group on day 7 (P=0.002), but the changes of the total Tau and pTau-396/404 level did not show an obvious correlation. Epothilone D induced persistent expression of GAP43 in the damaged axons, detectable even on day 28 of the experiment.@*CONCLUSION@#Epothilone D treatment can protect against TON in rats by promoting the survival of injured RGCs, enhancing Tau content in the surviving RGCs, reducing Tau accumulation in injured axons, and stimulating sustained regeneration of axons.

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.

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 ferulate-heparin-poloxamer thermosensitive hydrogels on nerve regeneration in rats with spinal cord injury / 中国药理学通报

Aim To investigate the effect of ferulate-heparin-poloxamer (SF-HP) thermosensitive hydrogels on nerve regeneration in ratswith spinal cord injury, and provide experimental basis for clinical application. Methods SD rats were randomly divided into five groups: sham group, SCI group, SCI + HP group, SCI + SF group and SCI + SF-HP group. Rats were performed moderate contusion injuries using a vascular clip for 2 min to establish SCI animal model, then rats were given BBB score and inclined plate scoring function test after SCI. BDA tracing was employed to observe the recovery of nerve conduction function. Moreover, immunohistochemical staining and Western blot-were used to measure GAP43, Nestin and GFAP levels. Results BBB score and inclined plane test score significantly increased in SF-HP treated group as compared with the model group (P < 0. 01). Staining data showed that the structure of spinal cord was void, and this phenomenon was reversed after SF-HP administration. Data showed that the level of GAP43 and Nestin increased after SF treatment (P < 0. 05), the expression of GFAP decreased (P <0. 05), and the treatment effect of SF-HP hydrogels were better than those of SF alone (P < 0. 0 5) . BDA tracing data showed that the number of positive neurons markedly increased and the repair of nerve conduction function was significantly improved in SF-HP hydrogel group compared with SF group. Conclusion HP hydrogels enhance the effect of SF on the nerve regeneration in damaged spinal cord in rats.

Effect of magnolol on hippocampal neuroplasticity in unpredictable chronic mild stress treated rats / 中草药

Objective To investigate the effect and mechanism of magnolol on hippocampal neuroplasticity in depression model rats. Methods In this study, depression model rats were prepared with unpredictable chronic mild stress (UCMS), and was given different doses of magnolol (20, 40 mg/kg) for 28 d. The rats in the positive control group were given fluoxetine (20 mg/kg) for 28 d. The ameliorative effects of magnolol on symptom of depression were investigated through behavior tests including open-field test, sucrose preference test, and forced-swimming test. The mRNA levels of Map-2, Gap43, and SYP in the hippocampus, cortex and striatum of rats in each group were detected by qRT-PCR, and the localization and expression of MAP-2, GAP43, and SYP in the hippocampus were observed by immunohistochemical staining analysis. The quantitative analysis of MAP-2, p-MAP-2, and p-ERK in hippocampus of rats in each group were further analyzed by Western blotting. Results UCMS was able to decrease the sucrose preference index, reduce locomotor activity and increase the immobility time in the forced swimming test. Compared with model group, magnolol significantly increased the spontaneous activity of rats, increased the consumption of sugar and water, and decreased the immobility time of chronic stress rats in forced swimming (P < 0.05, 0.01). Magnolol reversed MAP-2 mRNA and protein level in the hippocampus, increased phosphorylated MAP-2 expression in the hippocampus (P < 0.05), and significantly restored the p-ERK expression (P < 0.05). Conclusion Magnolol can affect the phosphorylation of MAP-2 through ERK pathway and increase the expression of MAP-2, thus affecting the neuronal plasticity and exerting its antidepressant effect.

The effects of paired associative stimulation on sensorimotor function and the expression of MAP-2 and GAP-43 after focal cerebral ischemia and reperfasion / 中华物理医学与康复杂志

Objective To observe the effect of paired associative stimulation ( PAS) on the recovery of sensorimotor function and to explore the mechanism in terms of neural plasticity. Methods Ninety male adult Sprague-Dawley rats were randomly divided into a sham operation group (Sham group), a model group (Model group) and a paired associative stimulation group ( PAS group) , each of 30. Each group was then subdivided into 7-, 14-and 28-day subgroups with 10 rats in each. A model of focal cerebral ischemia and reperfusion was estab-lished using the Longa suture method in the Model and PAS groups. The rats in the Sham group underwent the same surgical procedure except for the occlusion of the middle cerebral artery. The rats received 30 minutes of paired pe-ripheral nerve stimulation and transcranial magnetic stimulation comprising 90 pairs at 0.05 Hz beginning 24 h after the occlusion. The impulse wave width of the peripheral nerve stimulation was 200 μs and the intensity was 6 mA. The intensity of the transcranial magnetic stimulation was 120％ of the resting motor threshold. The other two groups weren't given any intervention. Neurological function was tested using Garcia scores on the 1st, 7th, 14th and 28th day after surgery. The rats were then sacrificed and the expression of MAP-2 and GAP-43 in the ischemic penumbra were detected using western blotting and immunohistochemistry. Results No neurological dysfunction was ob-served in the Sham group at any time. Compared with the Sham group at the same time points, the average Garcia scores of the Model and PAS groups were significantly lower (P≤0.05). However, the average Garcia scores on the 7th, 14th and 28th day were significantly higher in the PAS group compared with the Model group at the same time points ( P≤0.05) . The average Garcia scores of the Model and PAS groups on the 28th day after surgery were significantly higher than those on the 1st day (P≤0.05), but only the PAS group's average Garcia score on the 28th day was significantly higher than that on the 7th day. Compared with the Sham group at the same time points, the expression of MAP-2 and GAP-43 protein in the Model and PAS groups was significantly higher, but with that of the Model group significantly lower than that of the PAS group ( all P≤0.05) . The protein expression of MAP-2 and GAP-43 protein in the PAS group on the 14th day was significantly higher than on the 7th and 28th day ( P≤0.05 for both) . Conclusions PAS can promote the recovery of sensorimotor function after cerebral thrombosis, at least in rats. That may be due to its promoting the expression of the neuroplasticity-associated proteins MAP-2 and GAP-43 in the ischemic penumbra.

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.

Study of acupuncture combined with rat nerve growth factor on neurobehavioral ability of cerebral palsy infant rats and its brain tissue growth and metabolism associated proteins / 中国针灸

<p><b>OBJECTIVE</b>To study the effects of acupuncture combined with rat nerve growth factor (NGF) on the cerebral palsy infant rats and the proteins which associated with growth, apoptosis and metabolism.</p><p><b>METHODS</b>Seventy infant rats were selected, Fifty infant rats of which were made the cerebral palsy infant model by the ligation of unilateral carotid artery for cerebral ischemia and oxygen-deficient environment, then the 30 model rats were randomly divided into a model group, a NGF group and a combined group, 10 rats in each group. Twenty infant rats were used in the sham-operated group and the blank control group, 10 rats in each group. The treatment was not given in the blank control group. The rats in the sham-operated group were cut the neck skin and separated the left carotid artery, and then sutured and disinfected the wound. The intraperitoneal injection of NGF (2000 U•kg•d) was used in the NGF group. Based on the injection in the NGF group, acupuncture was used in the combined group, once a day, and the acupoints were "Baihui" (GV 20), left "nieⅠ" (extra), "Dazhui" (GV 14), "Jizhong" (extra), "Quchi" (LI 11), "Yongquan" (KI 1), "Hegu" (LI 4), "Zhoujie" (extra) and "Xiqianxue" (extra). The same volume of saline was intraperitoneally injected in the model group for continuous 14 days. Neurobehavioral ability score was evaluated after treatment. TUNEL were conducted to detect the brain cell apoptosis rate and the expressions of apoptosis associated gene Bax, Bcl-2 and Casp3 were detected by PCR. The level of nerve growth associated protein (GAP-43) and energy metabolism-related protein monocarboxylate transporter protien 1(MCT 1) were detected by Western blot.</p><p><b>RESULTS</b>After intervention, the neurobehavioral ability of baby rats in the blank control and sham-operated group was normal, but there was various degrees of abnormity in the model group, NGF group and combined group. The scores of neurobehavioral ability of the combined group and NGF group were better than those of the model control (all <0.05), and the scores in the combined group was better than those in the NGF group (all <0.05). The left brain cell apoptosis rate, expressions of Bax and Casp3 in the combined group and NGF group were lower and the expressions of Bcl-2 were higher than those of the model group (all <0.05), with more obvious results of Bax and Gasp3 in the combined group than those in the NGF group (all <0.05). The protein levels of GAP-43 and MCT 1 in the combined group and NGF group were higher than those in the model group (all <0.05), with higher expressions in the combined group compared with those in the NGF group (both <0.05).</p><p><b>CONCLUSION</b>Acupuncture combined with NGF could improve the neurobehavioral ability of cerebral palsy infant rats, inhibit the nerve cell apoptosis and improve the brain tissue injure and energy metabolism by up-regulating the expressions of GAP-43 and MCT 1.</p>

Role of cPKCγ/GAP-43 signaling pathway in ketamine-induced apoptosis in hippocampal neurons of developing rats:an in vitro experiment / 中华麻醉学杂志

Objective To evaluate the role of conventional protein kinase Cγ (cPKCγ)/growthassociated protein-43 (GAP-43) signaling pathway in ketamine-induced apoptosis in hippocampal neurons of developing rats in an in vitro experiment.Methods Primarily cultured hippocampal neurons were seeded in culture plates at a density of 1×10.6 cells/ml and divided into 2 groups (n=10 each) using a random number table:control group (C group) and ketamine group (K group).Group C received no treatment.Ketamine was added with the final concentration of 300 μmol/L in group K.At 12 h of culture or incubation,the apoptosis in hippocampal neurons was detected by flow cytometry.The apoptotic rate was calculated.The expression of cPKCγ,GAP-43 and phosphorylated GAP-43 in hippocampal neurons was measured by Western blot.Results Compared with group C,the apoptotic rates of hippocampal neurons were significantly increased,and the expression of cPKCγ,GAP-43 and phosphorylated GAP-43 was down-regulated in group K (P<0.01).Conclusion The mechanism by which ketamine induces apoptosis in hippocampal neurons of developing rats may be related to inhibition of cPKCγ/GAP-43 signaling pathway activation in an in vitro experiment.

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.

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>

Is There Additive Therapeutic Effect When GCSF Combined with Adipose-Derived Stem Cell in a Rat Model of Acute Spinal Cord Injury?

OBJECTIVE: Functional and neural tissue recovery has been reported in many animal studies conducted with stem cells. However, the combined effect of cytokines and stem cells has not yet been adequately researched. Here, we analyzed the additive effects of granulocyte colony-stimulating factor (GCSF) on adipose-derived stem cells (ADSCs) infusion in the treatment of acute spinal cord injury (SCI) in rats. METHODS: Four days after intrathecal infusion tubes implantation in Sprague-Dawley rats, SCI was induced with an infinite horizon impactor. In the Sham group (n=5), phosphate-buffered saline was injected 3, 7, and 14 days after SCI. GCSF, ADSCs, and ADSCs with GCSF were injected at the same time in the GCSF (n=8), ADSC (n=8), and ADSC+GCSF groups (n=7), respectively. RESULTS: The ADSC and ADSC+GCSF groups, but not the GCSF group, showed significantly higher Basso-Beattie-Bresnahan scores than the Sham group during 8 weeks (p<0.01), but no significant difference between the ADSC and ADSC+GCSF groups. In the ladder rung test, all four groups were significantly different from each other, with the ADSC+GCSF group showing the best improvement (p<0.01). On immunofluorescent staining (GAP43, MAP2), western blotting (GAP43), and reverse transcription polymerase chain reaction (GAP43, nerve growth factor), the ADSC and ADSC+GCSF groups showed higher levels than the Sham and GCSF groups. CONCLUSION: Our analyses suggest that the combination of GCSF and ADSCs infusions in acute SCI in the rat does not have a significant additive effect. Hence, when combination agents for SCI stem cell therapy are considered, molecules other than GCSF, or modifications to the methodology, should be investigated.

Effect of exhaustive exercise preconditioning on GAP-43 and Nogo-A in rats with cerebral ischemia reper-fusion / 中华行为医学与脑科学杂志

Objective To observe the effect of exhaustive exercise preconditioning on GAP?43 and Nogo?A in rats with cerebral ischemia reperfusion. Methods 90 rats were randomly divided into sham oper?ation group,group of cerebral ischemia reperfusion(I/R) and exhaustive exercise preconditioning group.Rats were sacrificed at 6 h,1 d,3 d and 7 d respectively after injury. Neural functions were detected by shuttle box. Morphological changes of hippocampal neural cells were observed by HE staining. Expressions of GAP?43 and Nogo?A were detected respectively by immunohistochemistry and RT?PCR technology. Re?sults Compared with the sham group,the death rate of apoptotic neurons in I/R group was decreased( 6 h:(30.97±2.09)%,1 d:(38.41±1.10)%,3 d:(46.81±2.04)%,1 d:(43.46±1.57)%),the index of learning and memory ability(AARR?7 d:(38.00±12.60)%,PAL?7 d:(27.90±1.79)s) and expression of GAP?43 were decreased(6 h:(2.89±0.85),1 d:(4.06±0.25),3 d:(4.78±0.98),7 d:(7.02±0.21)),the expression of Nogo?A was increased(6 h:(2.93±0.19),1 d:(5.47±0.32),3 d:(4.62±0.26),7 d:(4.12±1.11))(P<0.05).Compared with I/R group,the death rate of apoptotic neurons in exhaustive exercise preconditioning group were decreased,the index of learning and memory ability(AARR?7 d:(20.66±7.60)%,PAL?7 d:(35.53±2.41)s) and expression of GAP?43 were decreased(6 h:(2.03±0.14),1 d:(2.92±0.27),3 d:(3.35±0.34),7 d:(5.24±0.52)),the expression of Nogo?A were increased(6 h:(3.92±0.51),1 d:(6.90± 0.79),3 d:(5.87±0.48),7 d:(5.37±0.50))(P<0.05). Conclusion Exhaustive exercise preconditioning ag? gravates the injury of neurons and neural function,which is related to the regulation of GAP?43 and Nogo?A in the hippocampus of rats.

Effect of testosterone on pathological aggression behavior and expression of postsynaptics density-95 and growth associated protein-43 in prefrontal cortex in puberty rats / 第二军医大学学报

Objective To explore the effect of serum testosterone level on the pathologically aggressive behavior and the synaptic plasticity in the prefrontal cortex of puberty rats after gonadectomy. Methods Thirty male rats, 21 days old, were randomly divided into 3 groups: gonadectomized group, model group and control group. The gonadectomized and model groups were given a series of standard stress from 21 days old to puberty to induce animal model of pathological aggressive behavior. Resident-intruder experiment was performed to observe the variation of aggressive behaviors of animals. Enzyme-linked immunosorbent assay was used to examine the serum testosterone level. Western blotting analysis was used to determine the expression of postsynaptics density-95 (PSD-95) and growth associated protein-43 (GAP-43) in the prefrontal cortex. Results Resident-intruder experiment showed that the latency to the first attack in the gonadectomized group decreased significantly (P<0.01,P<0.01) and the attack times after yield increased significantly compared with those in the other two groups (P<0.01,P<0.01). The serum testosterone in the gonadectomized group was significantly decreased compared with the other two groups as shown by ELISA results (P<0.01, P<0.01). In addition, the aggressive-related behavior indices had a moderate to high negative correlation with the testosterone level (P<0.01). Western blotting analysis showed that prefrontal cortex expression of PSD-95 and GAP-43 in the gonadectomized group was significantly lower than those in the other two groups(P<0.01, P<0.01). Conclusion Low serum testosterone level can cause damage to the neuroplasticity of prefrontal cortex in puberty rats, which might be related to the pathologically aggressive behavior.

Quercetin promotes neurite growth through enhancing intracellular cAMP level and GAP-43 expression / 中国天然药物

The present study was designed to investigate the role of quercetin on neurite growth in N1E-115 cells and the underlying mechanisms. Quercetin was evaluated for its effects on cell numbers of neurites, neurite length, intracellular cAMP content, and Gap-43 expression in N1E-115 cells in vitro by use of microscopy, LANCE(tm) cAMP 384 kit, and Western blot analysis, respectively. Our results showed that quercetin could increase the neurite length in a concentration-dependent manner, but had no effect on the numbers of cells. Quercetin significantly increased the expression of cellular cAMP in a time- and concentration-dependent manner. The Gap-43 expression was up-regulated in a time-dependent manner. In conclusion, quercetin could promote neurite growth through increasing the intracellular cAMP level and Gap-43 expression.

Influence of sevoflurane anesthesia on expression of GAP-43 and NCAM in hippocampal neurons of neonatal rats / 中华麻醉学杂志

Objective To evaluate the influence of sevoflurane anesthesia on the expression of growth-associated protein 43 (GAP-43) and neural cell adhesion molecule (NCAM) in hippocampal neurons of neonatal rats.Methods Thirty-six pathogen-free Sprague-Dawley rats,aged 7 weeks,weighing 15-20 g,were randomly divided into 4 groups (n =9 each) using a random number table:control group (C group),1.5％ sevoflurane 6 h group (L group),3％ sevoflurane 2 h group (H1 group) and 3％ sevoflurane 6 h group (H2 group).Group L inhaled 1.5％ sevoflurane in oxygen for 6 h.H1 and H2 groups inhaled 3％ sevoflurane in oxygen for 2 and 6 h,respectively.Group C inhaled 30％ oxygcn only.When the neonatal rats were 14 days old,the rats underwent Morris water maze test for 7 consecutive days.Place navigation and spatial probe tests were carried out.After the end of Morris water maze test,the rats were sacrificed,and the hippocampus was obtained for determination of the expression of GAP-43 and NCAM in hippocampal neurons.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,and the expression of GAP-43 was down-regulated in L,H1 and H2 groups,and the frequency of crossing the original platform was decreased in L and H2 groups.There was no significant difference in NCAM expression among the four groups.Conclusion The mechanism by which sevoflurane anesthesia decreases the cognitive function may be related to down-regulated expression of GAP-43,but not related to NCAM expression in hippocampal neurons of neonatal rats.

Ginkgo-damole injection combined with bone marrow mesenchymal stem cell transplantation improves neurologic recovery from cerebral infarction / 中国组织工程研究

BACKGROUND:Reconstruction of damaged brain tissue through cel transplantation has become a new way to treat cerebral infarction. In recent years, bone marrow mesenchymal stem cels have become the new darling in cel transplantation therapy. OBJECTIVE:To investigate the effect of ginkgo-damole injection combined with bone marrow mesenchymal stem cel transplantation to improve the neurological function of acute cerebral infarction rats and its mechanism. METHODS:Animal models of middle cerebral artery occlusion were made in rats using suture method, and then 60 rat models were randomly divided into control group, cel transplantation group and combination group. The control group was given intravenous injection of PBSvia the tail vein; the cel transplantation group was given intravenous injection of bone marrow mesenchymal stem cel suspension (2.5×109/L) via the tail vein; the combination group was given intravenous injection of bone marrow mesenchymal stem cel suspension (2.5×109 /L) and ginkgo-damole injection (2 mL/kg, once a day, totaly 5 days)via the tail vein. Modified neurological severity scores were recorded at 1, 3 days and 1, 2 weeks after transplantation. At 2 weeks after transplantation, expressions of brain-derived neurotrophic factor and growth associated protein 43 in the brain were detected using RT-PCR; cel apoptosis detected using MTT assay; BrdU positive cels counted using immunohistochemistry method. RESULTS AND CONCLUSION:There were no differences in the modified neurologic severity scores among the three groups at 1, 3 days after transplantation (P > 0.05), but the modified neurological severity scores in the combination group were lower than those in the cel transplantation group and control group at 1, 2 weeks after transplantation (P < 0.05). The expressions of brain-derived neurotrophic factor and growth associated protein 43 in the brain were significantly higher in the combination group than the other two groups at 2 weeks after transplantation (P < 0.05); compared with the other two groups, the number of apoptotic cels was less but the number of BrdU positive cels was higher in the combination group (P < 0.05). These findings indicate that the combination of ginkgo-damole injection and bone marrow mesenchymal stem cel transplantation can increase the expressions of brain-derived neurotrophic factor and growth associated protein 43 in the brain, inhibit cel apoptosis and improve neurological function in rats with cerebral infarction.

Expressions of hypothalamus BDNF and GAP-43 and their relation with behavior changes in response to restraint stress in rats / 中华创伤杂志

Objective To investigate the expressions of brain-derived nerve growth factor (BDNF) and growth-associated protein 43 (GAP-43) in the hypothalamus of rats inflicted with restraint stress and their relationship with behavioral changes.Methods Forty male SD rats were divided into control group,restraint stress 7-day group,restraint stress 14-day group,restraint stress 21-day group according to the random number table,with 10 rats per group.Behavior changes were observed by open-field test,serum levels of corticotrophin releasing hormone (CRH) by enzyme-linked immunosorbent assay,and expressions of BDNF and GAP-43 in the hypothalamus by western blotting.Results Restraint stress 7-day group exhibited increases in spanning lattice times (50.0 ± 7.0),standing times (11.4 ± 2.1)and modification times (11.2 ± 2.7) compared with all other groups (P ＜ 0.05).Restraint stress 14-day group and restraint stress 21-day group showed significant decreases in spanning lattice times (35.5 ±7.5,29.4 ± 6.8),standing times (7.8 ± 4.9,5.6 ± 3.9) and modification times (6.7 ± 2.9,4.4 ±2.6) compared with control group (42.6 ± 5.4,8.9 ± 4.3,and 7.9 ± 3.0) (P ＜ 0.05).Restraint 14-day and 21-day groups showed significant increases in serum CRH level [(750.73 ± 123.68) pg/ml and (793.06 ± 115.84)pg/ml] compared with that in restraint stress 7-day group [(500.48 ± 88.71)pg/ml,P ＜0.05],but all were lower than (336.72 ±45.34) pg/ml in control group (P ＜0.05).Levels of BDNF and GAP-43 in the hypothalamus were the lowest in control group (0.672 ± 0.185 and 0.694 ±0.253).However,restraint stress increased the expressions of BDNF and GAP-43 in the hypothalamus,with the highest level in restraint stress 21-day group (1.357 ± 0.524 and 1.486 ± 0.679) (P ＜ 0.05).Conclusion Restraim stress can up-regulate BDNF and GAP-43 proteins in the hypothalamus,and lead to plasticity changes that may relate to stress-related behavior.

Effect of Daidzein intravitreal injection on optic nerve injury in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To discuss Daidzein intravitreal injection whether has protective and recovery effects on acute nerve damages. METHODS:After the crush models of acute optic nerve were set up, 72 males SD rats were divided into 4 groups randomly as common group without surgery, FBS negative control group, Daidzein treatment group ( 10μmol/L, 100μmol/L, 1000μmol/L ) and positive control group using rats nerve growth factor ( mNGF, 100ng/mL ). Three days after interference, all experimental animals were executed. HE staining was used to evaluate morphologic change of the retina, immunohisochemical staining and western-blot tests for identifying and quantifying the distinct expression of Caspase-3 and GAP-43 among the groups. RESULTS: Compared with the normal group and negative control group, retinal morphology of different concentrations of each Daidzein treatment group and positive control group was more complete, the expression of Caspase-3 protein was relatively lower, the expression of GAP-43 protein was relatively higher, the differences have statistically significance (P<0. 05).CONCLUSION: Daizein injection in the vitreous cavity has the capacity of protection and restoration in rat's acute nerve damages.