[Effects of electroacupuncture on myelin-related protein in corpus callosum of rats with focal cerebral ischemia].

OBJECTIVE: To observe the effect of electroacupuncture(EA)on the expression of myelin basic protein (MBP), axon growth inhibitor Nogo-A and Nogo receptor (NgR) in corpus callosum of rats with focal cerebral ischemia, so as to explore the mechanism of EA underlying improving ischemic white matter injury. METHODS: Fourty male SD rats were randomly divided into normal, sham operation, model and EA groups, with 10 rats in each group. The focal cerebral ischemia rat model was established by occlusion of the middle cerebral artery (MCAO). EA was applied to "Baihui"(GV20) and "Zusanli"(ST36) on the left side for 30 min, once daily for 14 days. Neurological function score and the adhensive removal test were used to evaluate neurological deficit severity; Hematoxylin-esion staining was used to observe the pathological changes in myelin of corpus callosum and luxol fast blue（LFB） staining was used to observe the myelin of corpus callosum. Immunohistochemical staining and Western blot were used to detect the expressions of MBP、Nogo-A and NgR in the ischemic corpus callosum. RESULTS: After MCAO, the neurological function score was significantly increased (P<0.05), the time required for contact with tape and tape removal was longer (P<0.001), the intensity of LFB staining and the expression of MBP decreased, while the veside area and the expression of Nogo-A and its receptor NgR increased (P<0.01, P<0.05) in the model group relevant to the normal and sham operation groups. The fiber arrangement of the corpus callosum on the ischemic side was disordered and a large amount of myelin sheath was lost in the model group. Following the treatment, the neurological deficit score of EA group was gradually decreased and significantly decreased on the 3rd, 7th and 14th day (P<0.05), and the time to remove the adhesive tape was shortened at the 7th and 14th day (P<0.001). The shape of the corpus callosum in the EA group was close to normal, and the myelin structure was relatively complete. The intensity of LFB staining and the expression of MBP was increased (P<0.05, P<0.01) while the expression of Nogo-A and its receptor NgR were decreased in the EA group relevant to the model group (P<0.01). CONCLUSION: EA can play a protective role in myelin of the corpus callosum after cerebral ischemia, which may be related to down-regulating the expressions of Nogo-A and NgR.

Effects of Electroacupuncture on Expression of D1 Receptor (D1R), Phosphorylation of Extracellular-Regulated Protein Kinase 1/2 (p-ERK1/2), and c-Fos in the Insular Cortex of Ketamine-Addicted Rats.

BACKGROUND The aim of this study was to investigate the effects of electroacupuncture (EA) on expression of the D1 receptor (D1R), phosphorylation of extracellular-regulated protein kinase 1/2 (p-ERK1/2) and c-Fos in the insular cortex (IC) of ketamine-addicted rats. MATERIAL AND METHODS Sprague-Dawley rats were randomly divided into 7 groups: the normal group, the normal saline (NS) group, the ketamine (Ket) group, the U0126+Ket group, the SCH23390+Ket group, the Ket+acupoints EA (EA1) group, and the Ket+ non-acupoints EA (EA2) group. We used immunohistochemistry to detect the expression of D1R, p-ERK1/2, and c-Fos. We also used Nissl staining techniques to study the morphology of IC neurons. RESULTS Our study demonstrated that the ketamine group had sparsely distributed neurons, large intracellular vacuoles, nuclei shift, and unclear nucleolus. The number of Nissl-positive (neuronal) cells in the ketamine group were decreased than in the normal group. Our results also indicated that there was significantly lower expression of D1R, p-ERK1/2, and c-Fos in the IC of the U0126+Ket group, SCH23390+Ket group, and Ket+EA1 group as compared with that of the Ket group. CONCLUSIONS Ketamine addiction induces c-Fos overexpression in the IC by increasing the expression of D1R and p-ERK1/2. Acupoints EA downregulate D1R and p-ERK1/2 by reducing the overexpression of c-Fos.

[Effect of Electroacupuncture Intervention on Expressions of Nerve Growth Factor and Growth Arrest-specific Protein 7 in Ventral Posterolateral Thalamic Nucleus in Cerebral Ischemia Rats].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on expressions of nerve growth factor(NGF) and growth arrest-specific protein 7(Gas 7) in the right ventral posterolateral nucleus(VPL) in the focal cerebral ischemia (FCI) rats, so as to explore its possible mechanism underlying improvement of secondary injury of FCI. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into normal group, sham operation group, model group and electroacupuncture (EA) group (n=12 in each group) by random number table.The FCI model was made by occlusion of the right middle cerebral artery (MCAO) with thread embolus.One week after MCAO, EA(2 Hz,2 V) was performed on "Baihui"(GV 20) and left "Zusanli"(ST 36) for 30 min,once daily for successively 21 days.The expressions of NGF and Gas 7 in the right VPL were detected by immunohistochemistry (n=6 in each group) and Western blot (n=6 in each group), respectively;meanwhile Nissl staining was conducted to show VPL neurons. RESULTS: Nissl staining showed that the structure of right VPL was clear and complete,and the nuclei were centered and clear in the normal group and sham operation group;the VPL neurons were deeply stained, and the nuclei were pyknotic in the model group;the morphology of neurons in the EA group was similar to that of the normal group. The results of immunohistochemistry and Western blot were consistent. Expressions of NGF and Gas 7 proteins in the right VPL were not of significant differences between the sham operation group and the normal group (P>0.05). Compared with the normal group, the expressions of NGF and Gas 7 proteins in the VPL of the ischemic side were significantly increased in the model group(P<0.05). After treatment,the expression levels of NGF and Gas 7 proteins were further up-regulated in the EA group in comparison with the model group(P<0.05). CONCLUSIONS: The enhanced expressions of NGF and Gas 7 in the ischemic VPL of FCI rats may be involved in the neuroprotection and repairing; EA can significantly up-regulate the expressions of NGF and Gas 7 in VPL of the ischemic side, which may contribute to its effect in improving secondary thalamic impairment of FCI.

Effects of Polysaccharide of Gastrodia Elata Blume and Electro-Acupuncture on Expressions of Brain-Derived Neurotrophic Factor and Stem Cell Factor Protein in Caudate Putamen of Focal Cerebral Ischemia Rats.

BACKGROUND The aim of this study was to explore the neural protective effect of polysaccharide of Gastrodia elata Blume (PGB) and electro-acupuncture (EA) on focal cerebral ischemia rats. MATERIAL AND METHODS A total of 40 Sprague-Dawley rats were randomly divided into 5 groups (normal group, model group, PGB group, EA group and PGB+EA group). The model was prepared by middle cerebral artery occlusion (MCAO). Two week after modeling, rats were given PGB, EA, or a combination of the 2 in continuous treatment for 2 successive weeks. 14 days after modeling, expressions of BDNF and SCF protein in the caudate putamen (CPu) were detected by immunohistochemistry. RESULTS Positive expression of BDNF and SCF protein was found in the right caudate putamen of each group of rats. Expressions of BDNF and SCF in the CPu of the model group were higher than normal group (P<0.05). Compared with the model group, the expressions of BDNF and SCF in the CPu of the PGB group, the EA group, and the PGB plus EA group increased significantly (P<0.05). The expressions of BDNF and SCF obviously increased in the PGB plus EA group compared to those of the EA group and the PGB group (P<0.05). CONCLUSIONS PGB and EA up-regulated the expressions of BDNF and SCF protein in the CPu of focal cerebral ischemia rats, and the combination of PGB+EA has a synergistic effect on the recovery from cerebral ischemia.

[Effect of Electroacupuncture Combined with Polysaccharide of Gastrodia elata Blume on Ex- pression of Brain Derived Neurotrophic Factor and Vascular Endothelial Growth Factor in the Paraventricular Nucleus of Hypothalamus in Cerebral Ischemia Rat].

OBJECTIVE: To observe the effect of electroacupuncture(EA) of "Baihui" (GV 20) and "Zusanli" (ST 36) in combination with administration of polysaccharide of Gastrodia elata Blume (PGB) on the expression of brain derived neurotrophic factor (BDNF) and vascular endothelial growth factor(VEGF) in the paraventricular nucleus of hypothalamus(PVN) of cerebral ischemia (C) rats, so as to explore its mechanism underlying improvement of Cl. METHODS: Forty Sprague-Dawley rats were randomized into five groups: normal control, model, EA, PGB, and EA+ PGB (n = 8/group). The cerebral ischemia model was established by occlusion of the middle cerebral artery (MCAO). EA (2 Hz, 2 V) was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) for 30 min, once daily for 14 days. The rats of the PGB and PGB+ EA groups were treated by intragastric gavage of PGB at a dose of 100 mg/kg, once daily for 14 successive days. The expression of BDNF and VEGF in the PVN of hypothalamus was detected by immunohistochemistry. RESULTS: After MCAO, the BDNF and VEGF immunoreactions (IR) positive neuron numbers and expression level in the PVN of the hypothalamus were significantly increased in the model group (P<0.05) and were further up-regulated after administration of EA and PBG ( P<0. 05). The effects of EA+ PGB were evidently superior to those of simple EA and simple PBG in up-regulating BDNF and VEGF IR-positive neuron numbers and expression levels in the PVN ( P<0.05). CONCLUSION: EA combined with PGB can up-regulate the expression of BDNF and VEGF in the PVN of hypothalamus in cerebral ischemia rats, which might contribute to its effect in improving cerebral ischemia.

[Effect of Electroacupuncture Intervention Combined with Gastrodin Administration on Neurological Function, Nogo-A and NgR Expression in the Frontal Lobe Cortex of Focal Cerebral Ischemia Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention combined with medication (Gastrodin) on changes of neurological function and expression of Nogo-A and Nogo-A receptor (NgR) in the frontal lobe cortex around the ischemic loci of focal cerebral ischemia (FCI) rats, so as to explore its mechanism underlying improvement of neuroregeneration of FC. METHODS: Fifty male Sprague-Dawley rats were randomly divided into normal control, model, EA, medication and EA+ medication groups (n = 10 in each group). The FCI model was induced by occlusion of the middle cerebral artery (MCAO) with thread embolus. EA was applied to the left "Quchi" (LI 11) and "Hegu" (Li 4) for 30 min, once daily for 14 days after MCAO. For rats of the medication group, Gastrodin (10 mg/kg) was administrated by intraperitoneal injection, once daily for 14 days. The neurological impairment was assessed by Zea Longa's scoring. The expression of Nogo-A and NgR in the frontal lobe cortex around the ischemic loci was detected by immunohistochemistry. RESULTS: In comparison with the normal control group, cerebro- cortical Nogo-A and NgR expression levels of the model group vere significantly increased (P < 0.05). Compared with the model group, the Zea Longa's score and Nogo-A and NgR expression levels were evidently down-regulated in the EA, medication and EA + medication groups (P < 0.05). The Zea Longa's score and Nogo-A and NgR expression levels were significantly lower in the EA + medication group than in the EA and medication groups (P < 0.05). CONCLUSION: EA intervention and Gastrodin administration can down-regulate cerebro-cortical Nogo-A and NgR protein expression in FCI rats, which may contribute to their action in improving neurological impairment. The effect of EA+ Gastrodin is better than simple EA or Gastrodin treatment.

[Effect of Electroacupuncture plus Polysaccharide of Gastrodia elata Blume on Expression of Nestin and Brain Derived Neurotrophic Factor in the Basolateral Amygdala of Focal Cerebral Ischemia Rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with administration of Polysaccharide of Gastrodia elata Blume (PGB) on stem cell proliferation in the basolateral amygdala (BLA) of focal cerebral ischemia (FCI) rats. METHODS: A total of 40 Sprague-Dawley rats were randomly divided into normal control,model,EA, PGB and EA+PGB groups (n=8 rats in each group). The FCI model was established by occlusion of the right middle cerebral artery. EA (2 Hz, 3 V) was applied to "Baihui" (GV 20) and the left "Zusanli"(ST 36) for 30 min, once daily for 14 successive days. Rats of the PGB and EA+PGB groups were given with PGB (100 mg/kg) by gastrogavage, once daily, for14 successive days. The Zea-Longa's method was used to determine the animals' nerurological dysfunction score on day 1 and 14. The expressions of Nestin and brain derived neurotrophic factor (BDNF) of the right BLA were measured by immunohistochemical staining. RESULTS: Following modeling, the neurological score, and the numbers of Nestin- and BDNF-immunoreaction (IR)-positive cells were significantly increased in the model group (P<0.05). Compared with the model group, the neurological scores were considerably decreased in the EA, PGB and EA+PGB groups (P<0.05), and the numbers of Nestin- and BDNF-IR positive cells were further significantly increased in the 3 treatment groups (P<0.05), and the effect of EA+PGB was notably superior to that of simple EA and simple PGB in down-regulating neurological score and up-regulating Nestin and BDNF expression (P<0.05). CONCLUSIONS: Both EA and PGB interventions can improve FCI rats' neurological function, which may be related to their effects in up-regulating the expression of Nestin and BDNF to promote the proliferation of neural stem cells in the BLA.

[Effects of Electroacupuncture Intervention Combined with Gastrodin on Expression of Proteins Related to Proliferation-differentiation of Neural Stem Cells in Hippocampal CA 1 and CA 3 Regions in Focal Cerebral Ischemia Rats].

OBJECTIVE: To observe the effect of electroacupuncture(EA) combined with medication on changes of expression of Nestin, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) in the hippocampal CA 1 and CA 3 regions of focal cerebral ischemia (FC1) rats, so as to analyze its mechanisms underlying neuroprotection. METHODS: Fifty male SD rats were randomly divided into normal control, model, EA, medication, and EA+ medication groups (n = 10 in each group). The FCI model was established by middle cerebral artery occlusion (MCAO) with thread embolus. EA (2 Hz, 2 V) was applied to the left "Hegu"(LI 4) and "Quchi" (LI 11) for 30 min, once daily for 14 days after MCAO. Rats of the medication group were given with intraperitoneal injection of gastrodin (10 mg/kg). The expression of Nestin, GFAP and NSE in the hippocampal CA 1 and CA 3 regions were detected by immunohistochemistry. RESULTS: Compared with the normal control group, the numbers of Nestin- and GFAP-immunoreaction (IR) positive cells in both CA 1 and CA 3 regions of the hippocampus were significantly increased in the model ciroup (P<0.05), while those of NSE-IR positive cells in both CA 1 and CA 3 regions were significantly decreased in the mdlgroup (P<0.05). After EA and medication interventions, the numbers of Nestin- and NSE-IR positive cells in the CA 1 and CA 3 regions were evidently increased and GFAP-IR positive neurons were considerably reduced in the EA, medication and EA+ medication groups (P<0.05). The effects of EA+ medication were significantly superior to those of both EA and simple medication in up-regulating the number of Nestin- and NSE-IR positive cells and down-regulating the number of GFAP positive neurons in CA 1 and CA 3-regions (P<0.05). CONCLUSION: EA and EA intervention combined with gastrodin can significantly up-regulate the number of Nestin- and NSE-IR positive cells, and down-regulate the number of GFAP positive cells in the CA 1 and CA 3 regions of hippocampus in focal cerebral ischemia rats, which may contribute to their effects in promoting the differentiation and proliferation of mature neurons in the hippocampus for improving cerebral functions. The effects of EA+ medication are obviously better than simple EA intervention.

[Effects of electroacupuncture intervention combined with polysaccharide of Gastrodia elata Blume on expression of nestin and cytokines of neural stem cells in the dentate gyrus of cerebral ischemia rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Baihui" (GV 20) and "Zusanli" (ST 36) and EA plus polysaccharide of Gastrodia elata Blume (PGB) on neural stem cell (NSC) proliferation in cerebral ischemia (CI) rats, so as to explore its mechanisms underlying improvement of CI. METHODS: A total of 40 SD rats were randomly divided into normal control, CI model, EA , medication (PGB) and EA + medication groups (n = 8 in each group). The CI model was established by occlusion of the middle cerebral artery (MCAO). Two weeks after modeling, rats of the medication and EA + medication groups were administrated with PGB (100 mg/kg) once daily for 2 successive weeks. EA (2 Hz, 3 V) was applied to "Baihui" (GV 20) and left "Zusanli" (ST 36) for 30 min, once daily for 2 weeks. The expression of Nestin and stem cell factor (SCF) proteins in the dentate gyrus (DG) of hippocampus was detected by immunohistochemistry. RESULTS: Compared with the normal control group, the expression of Nestin and SCF proteins in the suprapyramidal blade (SPB) and infrapyramidal blade (IPB) of DG were significantly increased in the model group (P < 0.05). After EA intervention, the expression levels of Nestin and SCF of SPB and IPB of DG were considerably further up-regulated in both EA group and EA + medication group (P < 0.05), and the effect of EA + medication group was significantly superior to that of simple EA group and medication group (P < 0.05). CONCLUSION: EA combined with PGB can significantly up-regulate the expression of Nestin and SCF in the hippocampal DG in CI rats, and its effects are apparently superior to those of simple EA or simple medication. It suggests that EA-induced proliferation of SCF may be one of the mechanisms underlying relieving CI.