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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.
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Animals , Male , Rats , Electroacupuncture , GAP-43 Protein/genetics , Moxibustion , Rats, Sprague-Dawley , Sciatic Nerve , Sciatica/therapy , Spinal CordABSTRACT
<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<sup>-1</sup>), Gap26 was intraperitoneally injected once a day since the third day after surgery (25 g·kg<sup>-1</sup>). Brain was taken out at the 7<sup>th</sup> 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.
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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.
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Animals , Male , Mice , Coronary Artery Disease , Electroacupuncture , Mice, Inbred C57BL , Myocardial Ischemia , MyocardiumABSTRACT
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.
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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.
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<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>
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Neodiplostomum seoulense (Digenea: Neodiplostomidae) is an intestinal trematode that can cause severe mucosal pathology in the small intestines of mice and even mortality of the infected mice within 28 days after infection. We observed neuronal growth associated protein-43 (GAP-43) expression in the myenteric plexus of the small intestinal wall of N. seoulense-infected mice until day 35 post-infection (PI). BALB/c mice were infected with 200 or 500 N. seoulense metacercariae isolated from naturally infected snakes and were killed every 7 days for immunohistochemical demonstration of GAP-43 in the small intestines. N. seoulense-infected mice showed remarkable dilatation of intestinal loops compared with control mice through days 7-28 PI. Conversely, GAP-43 expression in the mucosal myenteric plexus was markedly (P<0.05) reduced in the small intestines of N. seoulense-infected mice during days 7-28 PI and was slightly normalized at day 35 PI. From this study, it is evident that neuronal damage occurs in the intestinal mucosa of N. seoulense-infected mice. However, the correlation between intestinal pathology, including the loop dilatation, and depressed GAP-43 expression remains to be elucidated.
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Animals , Female , Humans , Male , Mice , Down-Regulation , GAP-43 Protein/genetics , Intestine, Small/metabolism , Metacercariae/growth & development , Mice, Inbred BALB C , Mice, Inbred C3H , Trematoda/growth & development , Trematode Infections/geneticsABSTRACT
@#Objective To explore the application values of effect on the expression of growth associated protein-43 (GAP-43) and myelin basic protein(MBP) by electrical stimulation in different sites of spinal cord injured rats after the human umbilical cord blood stem cells transplantation. Methods In this study the subjects adopted are clean grade rats, which were performed with Allen's method by NYU blow device, resulting in SCI models. 192 rats successfully modeled were divided into groups according to the random table. Group A received electric stimulation in the scalp surface projection area of motor area: Group A1 received electric stimulation and transplantation of umbilical cord blood stem cell, while Group A2 only received electric stimulation; Group B received local electric stimulation at damaged site: Group B1 received electric stimulation and transplantation of umbilical cord blood stem cell, while Group B2 only received electric stimulation; Group C received electric stimulation in the scalp surface projection area of motor area and local electric stimulation at damaged site: Group C1 received electric stimulation and transplantation of umbilical cord blood stem cell, while Group C2 only received electric stimulation; Group D are just the SCI models: Group D1 received transplantation of umbilical cord blood stem cell without electric stimulation, while Group D2 neither received cell transplantation nor electric stimulation. The sacrifice time of rats was the 1st week, 2nd week, 3rd week, 4th week, 8th week, and 12th week after operation respectively. The distribution of the remained or regenerative neurons at damaged areas by corticospinal tract anterograde tracer with Biotinylated dextran amine (BDA), and we also detected the content changes of GAP-43, MBP and the MAB1281 specific expression of the human nucleus of human umbilical cord blood stem cells. Results 1). NYU is a blow device for Allen's rat model of SCI, which could quantify the crash potential energy, and gain a stable and repeated model with high successful rate. 2). At each time site in this study, the factors that benefit for the neurofunction restoration after electric stimulation in the microenvironment into which the stem cells have been transplanted are GAP-43 and MBP, and they are all positive; the effects of the combination of head acupuncture with body acupuncture are much better than single head acupuncture or body acupuncture. 3). At each time site in this study, the factors that benefit for the neurofunction restoration after injury are GAP-43 and MBP, and the effects of them are all positive, which do not depend on whether the stem cell transplantation has been done. Conclusion 1). After the stem cell transplantation have been done, the changes of the microenvironment develops in the direction of benefiting for restoring neurofunction, which means that stem cell transplantation can promote the restoration of neurofunction and the stem cell transplantation is successful in this study. 2). Electric stimulation is significantly positive for the factors (GAP-43, MBP) which benefit for restoring neurofunction in the microenvironment after stem cell transplantation, the effects of the combination of head acupuncture with body acupuncture is better than single head acupuncture or body acupuncture. The significantly positive effects of the electric stimulation on microenvironment after injuries are independent, which do not depend on whether the stem cell transplantation has been done. Electric stimulation and stem cell transplantation are significantly synergistic at the microenvironment.
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@# Objective To explore the effects of electrical stimulation in different acupoints on the expression of growth associated protein-43 (GAP-43), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) in spinal cord injured rats after the human umbilical cord blood stem cells transplantation.Methods 192 SD rats were injured at T10-11 level with NYU, and divided randomly into groups: Group A received electric stimulation in the scalp of motor area (A1 group received electric stimulation and transplantation of umbilical cord blood stem cell, the rats of A2 group only received electric stimulation); group B received local electric stimulation at damaged site (B1 with electric stimulation and transplantation, B2 only with electric stimulation); group C received electric stimulation in the scalp of motor area and at damaged site (C1 with electric stimulation and transplantation, C2 only with electric stimulation); D1 received transplantation without electric stimulation, D2 neither with transplantation nor electric stimulation. The expression of GAP-43, bFGF and IGF were detected 1, 2, 3, 4, 8 and 12 weeks after operation. Results Electric stimulation increased the expression of GAP-43, bFGF, IGF, stimulating scalp and body were more than stimulated scalp or body alone, combined with the stem cells transplantation were more than no transplantation. Conclusion Both electric stimulation and stem cells transplantation can improve the microenvironment for neural recovery synergistically. Stimulating scalp and body is more effective than alone.
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@#Objective To explore the effect of repetitive transcranial magnetic stimulation(rTMS)on spinal cord injured rats.Methods Weight-drop spinal cord injury model was made at thoracic 10 segments with NYU impactor device.Stimulated group received daily superthreshold rTMS continued for 4 weeks.BBB locomotor scores were recorded weekly.Growth associated protein 43(GAP43)and 5-hydroxytryptamine(5-HT)were detected with immunofluorescence staining in the area of rostral and caudal to the lesion.Results The BBB scores in stimulation group improved compared with that in the control(P<0.01).GAP43 and 5-HT markers increased in the stimulation group(P<0.01),and they increased in the rostral than in the caudal areas(P<0.01).Conclusion rTMS can improve the locomotor function of incomplete spinal cord injury rats,which may result from the increase of expression of GAP43 and 5-HT.
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@#ObjectiveTo investigate the effect of Schwann cells (SCs) transplanted into cerebral hemorrhage area on never restore in rat.MethodsSCs were expended and labeled with 5'-Bromodexyuridine (BrdU) in vitro, then transplanted into cerebral hemorrhage area of model rat. Double immunohistochemistry staining was used to detect the expression of BrdU/myelin basic protein (MBP) and BrdU/growth associated protein-43 (GAP-43) respectively.ResultsBrdU/MBP positive cells could be seen one week after transplantation and up to 13 weeks. GAP-43 positive cells appeared in 12 weeks and 13 weeks, which was more in Hippocamp.ConclusionGrafted SCs can participate in remyelination and promoter nerve restore.
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The loss of neurons and synaptic contacts following cerebral ischemia may lead to a synaptic plastic modification, which may contribute to the functional recovery after a brain lesion. Using synapsin I and GAP-43 as markers, we investigated the neuronal cell death and the synaptic plastic modification in the rat hippocampus of a middle cerebral artery occlusion (MCAO) model. Cresyl violet staining revealed that neuronal cell damage occurred after 2 h of MCAO, which progressed during reperfusion for 2 weeks. The immunoreactivity of synapsin I and GAP-43 was increased in the stratum lucidum in the CA3 subfield as well as in the inner and outer molecular layers of dentate gyrus in the hippocampus at reperfusion for 2 weeks. The immunoreactivity of phosphosynapsin was increased in the stratum lucidum in the CA3 subfield during reperfusion for 1 week. Our data suggest that the increase in the synapsin I and GAP-43 immunoreactivity probably mediates either the functional adaptation of the neurons through reactive synaptogenesis from the pre-existing presynaptic nerve terminals or the structural remodeling of their axonal connections in the areas with ischemic loss of target cells. Furthermore, phosphosynapsin may play some role in the synaptic plastic adaptations before or during reactive synaptogenesis after the MCAO.