Electroacupuncture alleviates orofacial allodynia and anxiety-like behaviors by regulating synaptic plasticity of the CA1 hippocampal region in a mouse model of trigeminal neuralgia

OBJECTIVE To investigate whether electroacupuncture(EA)ameliorates abnormal trigeminal neuralgia(TN)orofacial pain and anxiety-like behavior by altering synaptic plasticity in the hippocampus CA1.METHODS A mouse infraorbital nerve transection model(pTION)of neuropathic pain was established,and EA or sham EA was used to treat ipsilateral acu⁃puncture points(GV20-Baihui and ST7-Xia⁃guan).Golgi-Cox staining and transmission elec⁃tron microscopy(TEM)were administrated to observe the changes of synaptic plasticity in the hippocampus CA1.RESULTS Stable and persistent orofacial allodynia and anxiety-like behav⁃iors induced by pT-ION were related to changes in hippocampal synaptic plasticity.Golgi stain⁃ings showed a decrease in the density of dendritic spines,especially mushroom-type dendritic spines,in hippocampal CA1 neurons of pT-ION mice.TEM results showed that the density of synapses,membrane thickness of the postsynaptic density,and length of the synaptic active zone were decreased,whereas the width of the synaptic cleft was increased in pTION mice.EA attenu⁃ated pT-ION-induced orofacial allodynia and anx⁃iety-like behaviors and effectively reversed the abnormal changes in dendritic spines and syn⁃apse of the hippocampal CA1 region.CONCLU⁃SION EA modulates synaptic plasticity of hippo⁃campal CA1 neurons,and reduces abnormal oro⁃facial pain and anxiety-like behavior,providing evidence for a TN treatment strategy.

Activation of metabotropic glutamate receptor 1 regulates hippocampal CA1 region excitability in rats with status epilepticus by suppressing the HCN1 channel

Dysregulation of hyperpolarization-activated cyclic nucleotide-gated cation(HCN)channels alters neuronal excitability.However,the role of HCN channels in status epilepticus is not fully understood.In this study,we established rat models of pentylenetetrazole-induced status epilepticus.We performed western blot assays and immunofluorescence staining.Our results showed that HCN1 channel protein expression,particularly HCN1 surface protein,was significantly decreased in the hippocampal CA1 region,whereas the expression of HCN2 channel protein was unchanged.Moreover,metabolic glutamate receptor 1(mGluR1)protein expression was increased after status epilepticus.The mGluR1 agonist(RS)-3,5-dihydroxyphenylglycine injected intracerebroventricularly increased the sensitivity and severity of pentylenetetrazole-induced status epilepticus,whereas application of the mGluR1 antagonist(+)-2-methyl-4-carboxyphenylglycine(LY367385)alleviated the severity of pentylenetetrazole-induced status epilepticus.The results from double immunofluorescence labeling revealed that mGluR1 and HCN1 were co-localized in the CA1 region.Subsequently,a protein kinase A inhibitor(H89)administered intraperitoneally successfully reversed HCN1 channel inhibition,thereby suppressing the severity and prolonging the latency of pentylenetetrazole-induced status epilepticus.Furthermore,H89 reduced the level of mGluR1,downregulated cyclic adenosine monophosphate(cAMP)/protein kinase A expression,significantly increased tetratricopeptide repeat-containing Rab8b-interacting protein(TRIP8b)(1a-4)expression,and restored TRIP8b(1b-2)levels.TRIP8b(1a-4)and TRIP8b(1b-2)are subunits of Rab8b interacting protein that regulate HCN1 surface protein.

Characterization of astrocytes and microglial cells in the hippocampal CA1 region after transient focal cerebral ischemia in rats treated with Ilexonin A

Ilexonin A is a compound isolated from the root of Ilex pubescens,a traditional Chinese medicine.Ilexonin A has been shown to play a neuroprotective role by regulating the activation of astrocytes and microglia in the peri-infarct area after ischemia.However,the effects of ilexonin A on astrocytes and microglia in the infarct-free region of the hippocampal CA1 region remain unclear.Focal cerebral ischemia models were established by 2-hour occlusion of the middle cerebral artery in rats.Ilexonin A(20,40 or 80 mg/kg)was administered immediately after ischemia/reperfusion.The astrocyte marker glial fibrillary acidic protein,microglia marker Iba-1,neural stem cell marker nestin and inflammation markers were detected by immunohistochemistry and western blot assay.Expression levels of tumor necrosis factor-αand interleukin 1βwere determined by enzyme linked immunosorbent assay in the hippocampal CA1 tissue.Astrocytes were activated immediately in progressively increasing numbers from 1,3,to 7 days post-ischemia/reperfusion.The number of activated astrocytes further increased in the hippocampal CA1 region after treatment with ilexonin A.Microglial cells remained quiescent after ischemia/reperfusion,but became activated after treatment with ilexonin A.Ilexonin A enhanced nestin expression and reduced the expression of tumor necrosis factor-αand interleukin 1βin the hippocampus post-ischemia/reperfusion.The results of the present study suggest that ilexonin A has a neuroprotective effect in the hippocampus after ischemia/reperfusion,probably through regulating astrocytes and microglia activation,promoting neuronal stem cell proliferation and reducing the levels of pro-inflammatory factors.This study was approved by the Animal Ethics Committee of the Fujian Medical University Union Hospital,China.

Atorvastatin increases dynamin 1 expression in hippocampal CA1 region in a rat model of vascular dementia

The current study examined a rat model of vascular dementia. The model rats exhibited obvious morphological and ultrastructural changes in neurons in the brain, and significantly reduced dynamin 1 expression in hippocampal CA1 region along with decreased learning and memory performance. Following atorvastatin treatment, the morphology and ultrastructure of cells in the model rat brain were significantly improved, dynamin 1 expression in hippocampal CA1 region was significantly enhanced, and learning and memory ability was significantly improved. The results demonstrated that impaired learning and memory abilities in vascular dementia model rats were closely correlated with decreased dynamin 1 expression. These findings indicate that atorvastatin can protect model rats against cognitive impairment by increasing dynamin 1 expression.

Somatosensory stimulation suppresses the excitability of pyramidal cells in the hippocampal CA1 region in rats

The hippocampal region of the brain is important for encoding environment inputs and memory formation. However, the underlying mechanisms are unclear. To investigate the behavior of indi-vidual neurons in response to somatosensory inputs in the hippocampal CA1 region, we recorded and analyzed changes in local ifeld potentials and the ifring rates of individual pyramidal cells and interneurons during tail clamping in urethane-anesthetized rats. We also explored the mechanisms underlying the neuronal responses. Somatosensory stimulation, in the form of tail clamping, chan-ged local ifeld potentials into theta rhythm-dominated waveforms, decreased the spike ifring of py-ramidal cells, and increased interneuron ifring. In addition, somatosensory stimulation attenuated orthodromic-evoked population spikes. These results suggest that somatosensory stimulation sup-presses the excitability of pyramidal cells in the hippocampal CA1 region. Increased inhibition by local interneurons might underlie this effect. These ifndings provide insight into the mechanisms of signal processing in the hippocampus and suggest that sensory stimulation might have thera-peutic potential for brain disorders associated with neuronal hyperexcitability.

Effects of repetitive transcranial magnetic stimulation on cognitive function and cholinergic activity in the rat hippocampus after vascular dementia

Repetitive transcranial magnetic stimulation （rTMS） is a non-invasive treatment that can enhance the recovery of neurological function after stroke. Whether it can similarly promote the recovery of cognitive function after vascular dementia remains unknown, In this study, a rat model for vascular dementia was established by the two-vessel occlusion method. Two days after injury, 30 pulses of rTMS were ad- ministered to each cerebral hemisphere at a frequency of 0.5 Hz and a magnetic field intensity of 1,33 T. The Morris water maze test was used to evaluate learning and memory function. The Karnovsky-Roots method was performed to determine the density of cholinergic neurons in the hippocampal CA1 region. Immunohistochemical staining was used to determine the number of brain-derived neurotroph- ic factor （BDNF）-immunoreactive cells in the hippocampal CA1 region, rTMS treatment for 30 days significantly improved learning and memory function, increased acetylcholinesterase and choline acetyltransferase activity, increased the density of cholinergic neurons, and increased the number of BDNF-immunoreactive cells. These results indicate that rTMS can ameliorate learning and memory deficiencies in rats with vascular dementia, The mechanism through which this occurs might be related to the promotion of BDNF expression and subsequent restoration of cholinergic system activity in hippocampal CA 1 region.

Effects of electroacupuncture on microcirculatory blood flow and glucose transporter function in the hippocampus

Nerve cell metabolism in post brain ischemia depends on increased microcirculation perfusion and transport function of microvascular endothelial cells. In the present study, a rat model of middle cerebral artery occlusion was established to investigate the influence of electroacupuncture （EA） on hippocampal CA1 cerebral blood flow and glucose transporter 1 （GLUT1） expression in the microvascular endothelial cells. Following EA at Neiguan （PC 6）, the cerebral blood flow in the ischemic hippocampal CA1 region was significantly elevated, the number and microvascular integrated absorbance of the GLUTl-positive cells were significantly increased, nerve cell damage was ameliorated, and GLUT1 protein expression in the ischemic hippocampus was significantly increased. Results demonstrate that EA increased the cerebral blood flow of the hippocampal CA1 region and improved the glucose transport function, thereby attenuating neuronal injuries.

Anomalous expression of chloride transporters in the sclerosed hippocampus of mesial temporal lobe epilepsy patients

The Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 regulate the levels of intracellular chloride in hippocampal cells. Impaired chloride transport by these proteins is thought to be involved in the pathophysiological mechanisms of mesial temporal lobe epilepsy. Imbalance in the relative expression of these two proteins can lead to a collapse of CI- homeostasis, resulting in a loss of gamma-aminobutyric acid-ergic inhibition and even epileptiform discharges. In this study, we investigated the expression of Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 in the sclerosed hippocampus of patients with mesial temporal lobe epilepsy, using western blot analysis and immunohistochemistry. Compared with the histologically normal hippocampus, the sclerosed hippocampus showed increased Na＋-K＋-Cl- cotransporter 1 expression and decreased K＋-CI- cotransporter 2 expression, especially in CA2 and the dentate gyrus. The change was more prominent for the Na＋-K＋-CI- cotransporter 1 than for the K＋-CI- cotransporter 2. These experimental findings indicate that the balance between intracellular and extracellular chloride may be disturbed in hippocampal sclerosis, contributing to the hyperexcitability underlying epileptic seizures. Changes in Na＋-K＋-CI-cotransporter 1 expression seems to be the main contributor. Our study may shed new light on possible therapies for patients with mesial temporal lobe epilepsy with hippocampal sclerosis.

Therapeutic effect of bone marrow mesenchymal stem cells on cold stress induced changes in the hippocampus of rats

The present study aims to evaluate the effect of bone marrow mesenchymal stem cells on cold stress induced neuronal changes in hippocampal CA1 region of Wistar rats. Bone marrow mes- enchymal stem cells were isolated from a 6-week-old Wistar rat. Bone marrow from adult femora and tibia was collected and mesenchymal stem cells were cultured in minimal essential medium containing 10% heat-inactivated fetal bovine serum and were sub-cultured. Passage 3 cells were analyzed by flow cytometry for positive expression of CD44 and CD90 and negative expression of CD45. Once CD44 and CD90 positive expression was achieved, the cells were cultured again to 90% confluence for later experiments. Twenty-four rats aged 8 weeks old were randomly and evenly divided into normal control, cold water swim stress （cold stress）, cold stress ＋ PBS （intra- venous infusion）, and cold stress ＋ bone marrow mesenchymal stem cells （1 x 106; intravenous infusion） groups. The total period of study was 60 days which included 1 month stress period followed by 1 month treatment. Behavioral functional test was performed during the entire study period. After treatment, rats were sacrificed for histological studies. Treatment with bone marrow mesenchymal stem cells significantly increased the number of neuronal cells in hippocampal CA 1 region. Adult bone marrow mesenchymal stem cells injected by intravenous administration show potential therapeutic effects in cognitive decline associated with stress-related lesions.

Delayed hippocampal neuronal death in young gerbil following transient global cerebral ischemia is related to higher and longer-term expression of p63 in the ischemic hippocampus

The tumor suppressor p63 is one of p53 family members and plays a vital role as a regulator of neuronal apoptosis in the development of the nervous system. However, the role of p63 in mature neuronal death has not been addressed yet. In this study, we first compared ischemia-induced effects on p63 expression in the hippocampal regions （CA1-3） between the young and adult gerbils subjected to 5 minutes of transient global cerebral ischemia. Neuronal death in the hippocampal CA1 region of young gerbils was significantly slow compared with that in the adult gerbils after transient global cerebral ischemia, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons in the sham-operated young group was significantly low compared with that in the sham-operated adult group, p63 immunoreactivity was apparently changed in ischemic hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. In the ischemia-operated adult groups, p63 immunoreactivity in the hippocampal CA1 pyramidal neurons was significantly decreased at 4 days post-ischemia; however, p63 immunoreactivity in the ischemia-operated young group was significantly higher than that in the ischemia-operated adult group. At 7 days post-ischemia, p63 immunoreactivity was decreased in the hippocampal CA1 pyramidal neurons in both ischemia-operated young and adult groups. Change patterns of p63 level in the hippocampal CA1 region of adult and young gerbils after ischemic damage were similar to those observed in the immunohistochemical results. These findings indicate that higher and longer-term expression of p63 in the hippocampal CA1 region of the young gerbils after ischemia/reperfusion may be related to more delayed neuronal death compared to that in the adults.

电针预处理对血管性痴呆大鼠神经细胞谷氨酸-NMDA受体信号转导通路的影响

目的:观察电针预处理对血管性痴呆(vascular dementia,VD)大鼠防治作用的可能机制。方法:将SD大鼠随机分为正常组、假手术组、模型组、电针预处理组。采用改良线栓法制备VD模型。电针预处理"百会""肾俞""后三里"穴,连续波,频率为100次/min,强度1 mA,每日1次,连续进行10 d。用比色法测定各组大鼠海马谷氨酸(Glu)含量的变化,原位杂交法测定海马N-甲基-D-天冬氨酸受体1(NMDAR 1)mRNA表达的变化。结果:模型组海马Glu含量、NMDAR 1 mRNA表达明显高于假手术组(P<0.01),电针预处理组海马Glu含量、NMDAR 1 mRNA表达明显低于模型组(P<0.05)。结论:电针预处理可降低VD鼠海马组织Glu含量,下调海马NMDAR 1的表达,因而有可能通过减轻Glu-NMDAR信号路径诱导的细胞凋亡发生,保护脑细胞。

尿毒症脑病的临床及影像学特征

目的：观察尿毒症脑病（ＵＥ）的临床表现特点，探讨痫性抽搐（ＥＴ）与颅脑影像学异常结果的关系。方法：①对４１例尿毒症脑病患者进行颅脑ＣＴ或磁共振（ＭＲ）扫描并与同期４１例其他原因做颅脑ＣＴ的患者作比较。②对ＣＲＦ模型大鼠进行脑组织病理检查。结果：①尿毒症脑病患者均有多种不同程度的神经系统异常表现。②异常脑部影像学表现为：４１例中３５例次显示脑萎缩，１７例次有低密度病灶，１０例次为髓质病变；对照组分别是９例次、１例次和１例次，２组有显著性差异。③ＣＲＦ大鼠脑组织光镜检查发现，大脑皮质中可见散在的神经细胞坏死灶，小血管管壁增厚，血管减少，颞叶海马ＣＡ１区锥体细胞变性、坏死明显。结论：所有尿毒症患者均有不同程度的中枢神经系统损害的临床表现；动物实验提供的病理学结果及影像学检查显示的脑萎缩和低密度病灶是产生上述高级精神活动异常的重要原因。

电针对氯胺酮成瘾大鼠海马CA1区酪氨酸羟化酶、c-fos表达的影响

目的:探讨电针对氯胺酮滥用成瘾戒毒治疗的物质基础。方法:将40只SD大鼠随机分为正常对照组、生理盐水(10 mL/kg)对照组、氯胺酮(100 mg/kg)组、氯胺酮(100 mg/kg)加电针组。按上述设定剂量,经腹腔注射给药,每天1次,连续7 d。氯胺酮加电针组于给药1周开始低频(2 Hz)电针交替刺激双侧"足三里"与"三阴交"穴位,每次30 min,每天1次,连续1周。采用免疫组织化学染色方法显示大鼠海马CA1区酪氨酸羟化酶(TH)、c-fos的表达。结果:与正常对照组和生理盐水对照组比,氯胺酮组大鼠海马CA1区TH、c-fos免疫反应阳性神经元的数目和阳性产物的表达水平明显增强(P<0.05,P<0.01)。与氯胺酮组比,氯胺酮加电针组TH、c-fos免疫反应阳性神经元的数目和阳性产物的表达显著减弱(P<0.01)。结论:电针"足三里""三阴交"可明显抑制氯胺酮成瘾引起的海马CA1区TH、c-fos表达的增强,提示电针可能通过抑制海马多巴胺神经元的活动改善氯胺酮成瘾。

间歇性低压低氧预处理对脑缺血大鼠海马p-p38 MAPK表达的影响

目的:本文旨在观察间歇性低压低氧(IH)预处理诱导脑缺血耐受过程中,大鼠海马CA1区磷酸化p38MAPK(p-p38 MAPK)的表达以及表达p-p38 MAPK的星形胶质细胞数量。方法:将30只健康雄性Wistar大鼠随机分为6组(n=5):假手术(sham)0 min组、IH+sham 0 min组、sham 7 d组、IH+sham 7 d组、损伤性缺血(Is)7 d组、IH+Is 7 d组。通过硫堇染色对各组大鼠海马CA1区锥体神经元进行神经病理学评价;免疫组织化学染色观察pp38 MAPK的表达;免疫荧光双标法观察表达p-p38 MAPK的星形胶质细胞数量。结果:IH预处理可以诱导脑缺血耐受,同时引起大鼠海马CA1区p-p38 MAPK的表达明显增加,且上调星形胶质细胞中p-p38 MAPK的表达。结论:低压低氧预处理促大鼠海马CA1区锥体神经元和星形胶质细胞中p-p38MAPK上调可能是IH预处理保护脑的一个途经。

中国株丙型肝炎病毒(HCV)感染猕猴后5'NTR—C区基因组的cDNA序列分析

从用于接种猕猴的受ＨＣＶ感染的２份献血员的血清，第一代猕猴感染ＨＣＶ１１个月的１＃食蟹猴血清和第二代猕猴受第一代猕猴ＨＣＶ血清感染ＨＣＶ３个月后的１４＃恒河猴血清，提取ＫＮＡ，用自行设计的ＨＣＶ５＇非编码区和核心区（５＇ＮＴＲ—Ｃ区）引物进行逆转录ＰＣＲ，将扩增的７７９ｂｐｃＤＮＡ片段克隆到ｐＵＣ１９质粒上；每一株挑３个克隆用双脱氧链终止法测定其序列并矫正偏差。４株ＨＣＶ的５＇ＮＴＲ—Ｃ区序列，原代人Ａ株（ＣＸ１）ｃＤＮＡ全长７７９ｂｐ，原代人Ｂ株（ＣＸ２）ｃＤＮＡ全长７７８ｂｐ，第一代猕猴株（ＣＸ３）ｃＤＮＡ全长７７６ｂｐ，第二代猕猴株（ＣＸ４）ｃＤＮＡ全长均为７７７ｂｐ，ＣＸ１株和ＣＸ４株均在５＇ＮＴＲｎｔ—２１６有—Ｃ的插入，ＣＸ３和ＣＸ４Ｃ区ｎｔ３８５—３８７处的３个硷基缺失；ＣＸ１株与ＣＸ２、ＣＸ３、ＣＸ４比较，同源性分别为９８．０７％。９６．１５％、９５．２５％：ＣＸ２与ＣＸ３、ＣＸ４的同源性分别为９６．２８％、９５．７６％；ＣＸ３与ＣＸ４的同源性为９７．５６％。由克隆的ＨＣＶＣ区ｃＤＮＡ推导的氨基酸序列，从ＨＣＶ的启始密码起，ＣＸ１株、ＣＸ２株序列全长１６８个氨基酸。

Acupuncture improves dendritic structure and spatial learning and memory ability of Alzheimer's disease mice

Acupuncture can improve the cognitive state of Alzheimer＇s disease, but its mechanism is not clear. Dendritic atrophy and synaptic loss in Alzheimer＇s disease brain are positively correlated with cognitive damage. Therefore, we speculated that the effect of acupuncture on improving cognitive function may be associated with reduced dendritic damage in the brain. Acupuncture at Qihai（CV6）, Zhongwan（CV12）, Danzhong（CV17）, bilateral Zusanli（ST36）, and bilateral Xuehai（SP10） acupoints was performed once a day（1-day rest after 6-day treatment） for 14 consecutive days. Senescence-accelerated mouse prone 8（SAMP8） mice without acupuncture and senescence-accelerated mouse resistant 1（SAMR1） mice were used as normal controls. After 14 days of treatment, spatial learning and memory ability of mice was assessed in each group using the Morris water maze. Dendritic changes of pyramidal cells in the hippocampal CA1 region were analyzed by quantitative Golgi staining. Our results showed that acupuncture shortened escape latency and lengthened retention time of the former platform quadrant in SAMP8 mice. Further, SAMP8 mice exhibited a significant increase in the number of apical and basal dendritic branches and total length of apical and basal dendrites after acupuncture. These results suggest that acupuncture improves spatial learning and memory ability of middle-aged SAMP8 mice by ameliorating dendritic structure.Acupuncture can improve the cognitive state of Alzheimer＇s disease, but its mechanism is not clear. Dendritic atrophy and synaptic loss in Alzheimer＇s disease brain are positively correlated with cognitive damage. Therefore, we speculated that the effect of acupuncture on im- proving cognitive function may be associated with reduced dendritic damage in the brain. Acupuncture at Qihai （CV6）, Zhongwan （CV 12）, Danzhong （CV17）, bilateral Zusanli （ST36）, and bilateral Xuehai （SP10） acupoints was performed once a day （1-day rest after 6-day treat- ment） for 14 consecutive days. Senescence-accelerated mouse prone 8 （SAMP8） mice without acupuncture and senescence-accelerated mouse resistant 1 （SAMR1） mice were used as normal controls. After 14 days of treatment, spatial learning and memory ability of mice was assessed in each group using the Morris water maze. Dendritic changes of pyramidal cells in the hippocampal CA1 region were analyzed by quantitative Golgi staining. Our results showed that acupuncture shortened escape latency and lengthened retention time of the former platform quadrant in SAMP8 mice. Further, SAMP8 mice exhibited a significant increase in the number of apical and basal dendritic branches and total length of apical and basal dendrites after acupuncture. These results suggest that acupuncture improves spatial learning and memory ability of middle-aged SAMP8 mice by ameliorating dendritic structure.

Changes of paired-pulse evoked responses during the development of epileptic activity in the hippocampus

Dysfunction of inhibitory synaptic transmission can destroy the balance between excitatory and inhibitory synaptic inputs in neurons,thereby inducing epileptic activity.The aim of the paper is to investigate the effects of successive excitatory inputs on the epileptic activity induced in the absence of inhibitions.Paired-pulse orthodromic and antidromic stimulations were used to test the changes in the evoked responses in the hippocampus.Picrotoxin(PTX),γ-aminobutyric acid(GABA) type A(GABA A) receptor antagonist,was added to block the inhibitory synaptic transmission and to establish the epileptic model.Extracellular evoked population spike(PS) was recorded in the CA1 region of the hippocampus.The results showed that the application of PTX induced a biphasic change in the paired-pulse ratio of PS amplitude.A short latency increase of the second PS(PS2) was later followed by a reappearance of PS2 depression.This type of depression was observed in both orthodromic and antidromic paired-pulse responses,whereas the GABAergic PS2 depression [called paired-pulse depression(PPD)] during baseline recordings only appeared in orthodromic-evoked responses.In addition,the depression duration at approximately 100 ms was consistent with a relative silent period observed within spontaneous burst discharges induced by prolonged application of PTX.In conclusion,the neurons may ignore the excitatory inputs and intrinsically generate bursts during epileptic activity.The depolarization block could be the mechanisms underlying the PPD in the absence of GABA A inhibitions.The distinct neuronal responses to stimulations during different epileptic stages may implicate the different antiepileptic effects of electrical stimulation.

磷脂酰肌醇-3-激酶/蛋白激酶B信号转导通路与针刺保护癫痫继发海马神经元损伤的关系

目的:观察针刺对戊四唑(PTZ)诱发癫痫大鼠海马神经元损伤的保护作用及该保护作用与磷脂酰肌醇-3-激酶(PI 3K)/蛋白激酶B(Akt)信号转导通路的关系。方法:SD大鼠120只随机分为5组:正常组、PTZ组、LY 294002组、针刺+LY 294002组、针刺组,每组24只。针刺组、针刺+LY 294002组针刺"百会""大椎"30min,共治疗5次。正常组、PTZ组、针刺组先侧脑室注射二甲基亚砜5μL,LY 294002组、针刺+LY 294002组侧脑室注射LY 294002(5μL),30min后正常组腹腔注射生理盐水2mL,其余各组腹腔注射PTZ(50mg/kg),于注射后4h与24h取脑组织。分别用HE染色法于光镜下观察海马结构改变,用电镜观察海马超微结构。结果:癫痫发作后4h光镜、电镜均可见大鼠海马神经元损伤,24h后损伤进一步加重。LY 294002组大鼠海马神经元损伤明显加重。针刺+LY 294002组大鼠海马神经元损伤明显,与LY294002组比较未见明显好转。针刺组大鼠海马神经元损伤明显减轻。结论:针刺具有明显保护癫痫继发脑神经元损伤作用,其保护作用与细胞内PI 3K/Akt信号转导通路有关。

针刺对戊四唑诱发惊厥大鼠海马葡萄糖调节蛋白78和C/EBP同源蛋白表达的影响

目的:观察针刺对惊厥大鼠海马神经元内葡萄糖调节蛋白78(Grp 78)和C/EBP同源蛋白(CHOP)蛋白表达的影响,探讨针刺抗惊厥的可能机制。方法:SD大鼠随机分为正常对照组(n=6)、模型组(n=18)、针刺组(n=18)。腹腔注射戊四唑(50mg/kg)复制惊厥大鼠模型。针刺组立即针刺"百会""大椎"30min。各组分别于造模后2、12、48h采用免疫组化法观察各组大鼠海马CA 1区Grp 78和CHOP蛋白表达情况。结果:模型组大鼠在惊厥发作2、12h海马CA 1区Grp 78蛋白表达与正常组比较明显增强(P<0.01);针刺组大鼠在惊厥发作12h和48h与模型组比较Grp 78蛋白表达显著增加(P<0.01,P<0.05)。在惊厥发作各个时段与正常组比较,模型组大鼠海马CA 1区CHOP蛋白表达明显上调(P<0.01);针刺组海马CA 1区CHOP蛋白阳性表达在惊厥发作各个时段与模型组比较明显减少(P<0.05,P<0.01)。结论:针刺能明显调节惊厥大鼠海马神经元Grp 78蛋白和CHOP蛋白的表达,从而起到保护惊厥性脑损伤作用。

电针对缺氧缺血性脑病幼鼠学习记忆能力的影响

目的:观察电针对缺氧缺血性脑病幼鼠学习记忆能力的影响,探讨电针对其神经元的作用机制。方法:将7d龄SD大鼠随机分为假手术组(n=12)、模型组(n=11)和电针组(n=12)。采用左颈总动脉结扎联合缺氧的方法建立缺氧缺血性脑病模型。电针组予电针"百会""大椎",隔日1次,每次20min,连续28d。观察治疗后幼鼠转棒跌落潜伏期、高架迷宫进入开放臂时间、水迷宫逃避潜伏期和逃避距离,高尔基染色法观察幼鼠神经元树突棘的密度。结果:转棒跌落潜伏期和高架迷宫进入开放臂时间,3组之间差异无统计学意义(P>0.05)。水迷宫实验,模型组较假手术组逃避潜伏期延长(P<0.05),逃避距离增加(P<0.05),平台象限停留时间减少(P<0.05),原平台穿越次数减少(P<0.05);电针组较模型组逃避潜伏期缩短(P<0.05),逃避距离减少(P<0.05),平台象限停留时间增加(P<0.05),原平台穿越次数增加(P<0.05)。模型组较假手术组树突棘密度降低(P<0.05),电针组较模型组树突棘密度升高(P<0.05)。结论:电针可以改善幼鼠缺氧缺血造成的学习记忆能力损伤,其作用可能与调节神经元树突棘密度有关。