经鼻给予TGFβ1对氯化锂-匹罗卡品诱导的SE大鼠海马神经元凋亡调控因子Bcl-2、Bax蛋白表达的影响

目的探讨经鼻给予TGFβ1(Transforming growth factor-beta1,TGFβ1)对氯化锂-匹罗卡品诱导的癫痫持续状态(status epilepticus,SE)大鼠海马神经元凋亡调控因子Bcl-2、Bax蛋白表达的影响。方法健康雄性SD大鼠60只,随机分为TGF组、Pilo组和正常对照组(Control)。建立氯化锂-匹罗卡品癫痫持续状态模型。采用免疫组化方法检测凋亡相关基因Bcl-2、Bax的蛋白表达。结果 (1)SE后24h、48h、72h,TGF组大鼠海马Bax阳性细胞均较Pilo组显著减少(P<0.05);72h最为明显(P<0.01)。HE染色是对各组大鼠海马神经元的形态结构变化的大体观察。(2)SE后24h、48h、72h,TGF组大鼠海马Bcl-2阳性细胞均较Pilo组显著增加(P<0.05);24h最为明显(P<0.01)。结论经鼻(IN)给予TGFβ1可以显著抑制癫痫持续状态诱导的大鼠海马神经元Bax蛋白的表达,上调Bcl-2蛋白表达,从而发挥神经保护作用。     

Effects of acupoint versus non-acupoint electroacupuncture on cerebral cortical neuronal Bcl-2, Bax and caspase-3 expression in a rat model of focal cerebral ischemia

BACKGROUND： Several studies have demonstrated that electroacupuncture by acupoint selection can inhibit cerebral cortical neuronal apoptosis following cerebral ischemia/reperfusion. OBJECTIVE： To validate the effects of electroacupuncture by acupoint selection on the expression level of cortical neuronal anti-apoptotic Bcl-2 protein and the apoptotic executive protein, caspase-3, in rat models of focal cerebral ischemia/reperfusion. DESIGN, TIME AND SETTING： This randomized grouping, neural cell and molecular biology animal experiment was performed at the Laboratory of Pharmacology of Traditional Chinese Medicine and the Laboratory Animal Center of Henan Institute of Traditional Chinese Medicine between November 2006 and May 2007. MATERIALS： Atotal of 40 healthy male adult Sprague-Dawley rats were randomly and evenly divided into four groups： sham-operated, model, electroacupuncture and non-aeupoint control. G6895 electro-acupuncture instruments were purchased from Shanghai Huayi Instrument Factory, China. Caspase-3, Bcl-2 and Bax kits were provided by Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Middle cerebral artery occlusion was induced in the model, electroacupuncture and non-acupoint groups. In the electroacupuncture group, the acupoints Jianyu （LII5）, Waiguan （S J5）, Biguan （ST31）, and Zusanli （ST36） were given electroacupuncture. In the non-acupoint control group, at each time point （immediately after ischemia and after reperfusion, or 2 hours after reperfusion）, electroacupuncture was performed at the midpoints of Tianquan （PC2）-Quze （PC 3） line, Quze （PC 3）-Ximen （PC4） line, Zuwuli （LR10）-Yinbao （LR9） line, and Xiguan （LR7）-Zhongdu （LR6） line. Electroacupuncture parameters were set with a continuous wave with a frequency of 10 Hz, wave width 0.6 ms, voltage 1.5-3.0 V, and a duration of 10 minutes. The sham-operated and model groups received only animal fixation without electroacupuncture procedure. MAIN OUTCOME MEASURES： Five ra     

Effects of acupuncture at Sibai （ST 2） acupoint on neuronal discharge in rat nucleus tractus solitarius

BACKGROUND： Electrophysiology can prove the integration of afferent information from the stomach meridian of Foot-Yangming in the nucleus tractus solitarius （NTS） and objectively describe the specific association between meridian vessels and Zang Fu organs. OBJECTIVE： To investigate the effects of afferent information from acupuncture at Sibai （ST 2） acupoint on neuronal discharge in rat NTS. DESIGN, TIME AND SETFING： A randomized, controlled, animal experiment was performed at the Key Laboratory of Meridian-Vessels and Zang Fu Organs, Traditional Chinese Medicine University of Hunan, State Administration of Traditional Chinese Medicine, and Key Laboratory of Acupuncture, Moxibustion, and the Biological Information of Hunan Higher Education Institutes, between December 2005 and October 2008. MATERIALS： A total of 52 Sprague Dawley rats, of either gender, aged 4 months, were included in this study. Acupuncture needles of 0.32 mm （diameter） x 40 mm （length） were used. METHODS： An extracellular recording protocol was applied. The Sibai （ST 2） acupoint in the stomach meridian of Foot-Yangming was used as an acupuncture point （acupoint）. Simultaneously, Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （S118） acupoint in the small intestine meridian of Hand-Taiyang, and a non-acupoint lateral to Sibai （ST 2） acupoint, were selected as controls. The Sibai （ST 2） acupoint was stimulated for 30 seconds by hand acupuncture through twirling and rotating, to determine the neurons responding to body surface stimulation in the NTS. MAIN OUTCOME MEASURES： Frequency of responding NTS neurons after acupuncture at four acupoints including Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44） and Quanliao （SI 18） and one non-acupoint. RESULTS： The frequency of responding NTS neurons was significantly higher after acupuncture at Sibaithan at control sites including the Dicang （ST 4）, Neiting （ST 44） and Quanliao （S118） acupoints and at the non-acupoint （P 〈 0.01）. The frequency of responding NTS neurons at Dicang （ST 4） and Quanliao （SI 18） was significantly higher than at Quanliao （SI 18） and the non-acupoint （P 〈 0.05）. The rate of frequency change of responding NTS neurons for the Sibai （ST 2）, Dicang （ST 4）, Neiting （ST 44）, and Quanliao （S118） acupoints as well as the non-acupoint was （35.08±4.80） %, （28.25± 5.46） %, （27.57± 4.87） %, （20.02 ±4.23） %, and （18.55 ±2.49） % respectively. Simultaneously, significant differences existed between Sibai （ST 2） and the other acupoints （P 〈 0.05 or P〈 0.01）. CONCLUSION： Compared with the Dicang （ST 4） and Neiting （ST 44） acupoints in the stomach meridian of Foot-Yangming, Quanliao （SI 18） acupoint in the small intestine meridian of Hand-Taiyang, and the non-acupoint lateral to Sibai （ST 2） acupoint, the Sibai （ST 2） acupoint in the stomach meridian of Foot- Yangming is more closely related to the NTS. In the stomach meridian of Foot- Yangming, afferent information is different in distant and near Shu acupoints, indicating that each Shu acupoint has its own specificity.     

Responses of neuronal nitric oxide synthase expression in the brainstem to electroacupuncture Zusanli (ST 36) in rats

Recent studies have reported that l-arginine-derived nitric oxide (NO) in the gracile nucleus modifies the hypotensive responses to electroacupuncture (EA) stimulation of Zusanli (ST 36). The purpose of this study was to examine the influence of EA stimulation of ST 36 on neuronal NO synthase (nNOS) expression in the brainstem nuclei in rats. EA stimulation of ST 36 and a non-acupoint was performed using 3 Hz of stimulation for 10 s every 2 min for a period of 120 min in rats anesthetized with ketamine. Rats in the sham-treated group received surgery and EA needles were placed into the acupoints without performing the stimulation. After 2-h stimulation and sham treatment, animals were perfused with 4% paraformaldehyde. Sections of rat medulla were examined by immunolabeling with a polyclonal antibody directed against nNOS. The brainstem nuclei were also visualized by NADPH-diaphorase histochemistry, a marker of nNOS activity. nNOS expression and NADPH-diaphorase reactivity were quantified by using a microscope with reticule grid to count the number of positive cells over a nucleus. Unilateral EA stimulation of ST 36 in rats caused increases in nNOS immunostained cells in the rostral region of the ipsilateral gracile nucleus, but was not altered in the contralateral gracile nucleus compared with sham-treated rats (P < 0.05, n = 6-7). NADPH-diaphorase-positive cells were also increased in the ipsilateral gracile nucleus of rats with EA stimulation. nNOS immunostaining and NADPH-diaphorase-positive neurons were significantly increased in both ipsilateral and contralateral sides of the medial nucleus tractus solitarius (mNTS) in rats receiving EA ST 36 compared with sham-treated animals (P < 0.05). nNOS immunostaining and NADPH-diaphorase reactivity was neither altered in the gracile nucleus and mNTS of non-acupoint stimulated rats nor other brainstem nuclei in rats with EA ST 36. These results show that nNOS immunoreactivity and NADPH-diaphorase reactivity are consistently increased in the gracile nucleus and the mNTS by EA ST 36. We conclude that EA ST 36 induces nNOS expression in the gracile nucleus and mNTS, and enhanced nNOS-NO in the nuclei may modify central cardiovascular regulation, which contribute to hypotensive effects of acupuncture.     

Effects of bFGF and TGFbeta on the expression of amyloid precursor and B-cell lymphoma protooncogene proteins in the rat retina

The pattern of immunoreactivity for amyloid precursor (APP) and B-cell lymphoma protooncogene (Bcl-2) proteins in the rat retina was studied after intravitreal injection of basic fibroblast growth factor (bFGF) or transforming growth factor-beta (TGFbeta). In normal control retinas, intense immunostaining of APP and Bcl-2 was observed primarily in the endfeet and proximal part of radial processes of Müller glial cells. A dose-dependent reduction in immunostaining of APP and Bcl-2 in Müller cells was observed after injection of bFGF and TGFbeta. These results provide the first evidence that APP and Bcl-2 can be down-regulated by cytokines in vivo.     

经鼻给予TGFβ1对癫痫持续状态大鼠海马神经元的保护作用

目的探讨经鼻腔给予TGFβ1(transforming growth factor beta1,TGFβ1)对氯化锂-匹罗卡品所致癫痫持续状态(status epilepticus,SE)大鼠海马神经元的保护作用及其潜在的机制。方法健康雄性SD大鼠60只,随机分为转化生长因子(TGF)组、匹罗卡品(Pilo)组和正常对照组(control)。建立氯化锂-匹罗卡品癫痫持续状态模型。应用TUNEL染色、Fluoro-Jade B(FJB)荧光染色法分别观察各组大鼠海马神经元的原位凋亡及变性死亡情况。采用免疫组化方法检测凋亡相关基因caspase-3的蛋白表达。结果 SE后24h、48h、72h,TGF组大鼠海马FJB、TUNEL、caspase-3阳性细胞均较Pilo组显著减少(P<0.05);72h最为明显(P<0.01)。结论经鼻(IN)给予TGFβ1可以显著抑制或减轻癫痫持续状态大鼠海马神经元的变性与凋亡,从而发挥神经保护作用。其潜在的神经保护机制可能涉及下调caspase-3蛋白表达。     

Roles of glutamate transporter and receptors, poly (ADPribose) polymerase, and transforming growth factor-beta1 in pontosubicular neuron necrosis

The expression of neuron-type glutamate transporters (EAAC-1), AMPA glutamate receptor subunits (GluR1 and GluR2/3), polyadenosine (5'diphosphate-ribose) polymerase (PARP), and transforming growth factor-beta1 was investigated in 20 cases of neonatal pontosubicular neuron necrosis and 12 gestational-age matched controls. Developmental immunoreactivities of EAAC-1, GluR1, and GluR2/3 appeared in the neurons of the pontine nuclei at 29 to 30 weeks' gestation in controls, and then gradually increased with age. However, these activities were decreased in the pontine nucleus of patients with pontosubicular neuron necrosis. Decreases in these immunoreactivities might indicate early degeneration of neurons. Although PARP and transforming growth factor-beta1 immunoreactivity was insignificant or very weak in the pontine nuclei at any age in controls, PARP was markedly expressed in karyorrhectic neurons of the pontine nucleus in patients with pontosubicular neuron necrosis. Transforming growth factor-beta1 immunoreactivity was observed in nonkaryorrhectic neurons of the pontine nuclei. PARP could contribute to the pathogenesis of pontosubicular neuron necrosis more than EAAC-1 or GluR1 or GluR2/3. Transforming growth factor-beta1 could play a role in the protection and repair of damaged neurons.     

Expression of transforming growth factor-beta 1 in periventricular leukomalacia.

The expression of transforming growth factor-beta 1, which has neurotrophic effects, was investigated in 25 neonates with periventricular leukomalacia using immunohistochemistry. In controls, transforming growth factor-beta 1 immunoreactivity was not detected in the cerebral white matter or cortex. Of the 25 cases of periventricular leukomalacia, transforming growth factor-beta 1 immunoreactivity was found in 16, and was distributed mainly in the cytoplasm of astrocytes, being prominent around necrotic foci in the white matter. The immunoreactivity was negative or weak at the acute stage of periventricular leukomalacia, and increased at the subacute stage and then decreased or was absent at the chronic stage. Astrocytes that were moderately or markedly positive for transforming growth factor-beta 1 were not found before 27 weeks' gestation, but were observed after 32 weeks' gestation in the white matter of the brains of neonates with periventricular leukomalacia. Transforming growth factor-beta 1 expression tended to be more obvious in focal periventricular leukomalacia than in widespread or diffuse periventricular leukomalacia. Our results suggest that transforming growth factor-beta 1 could be involved in the delayed glial response rather than the initial glial activation, and could play a role in the inhibition and repair of injury in periventricular leukomalacia. Exogenous transforming growth factor-beta 1 could have therapeutic potential for periventricular leukomalacia.     

Transforming growth factor-beta(1) restores antiplatelet function of endothelial cells exposed to anoxia-reoxygenation injury

Transforming growth factor-beta(1) released from platelet alpha-granules may preserve endothelial functions in injured vessels. However, direct evidence is lacking regarding how this cytokine modifies the antithrombotic function of injured endothelial cells. We performed an in vitro study to investigate the effects of transforming growth factor-beta(1) on platelet functions in the presence of cultured endothelial cells exposed to anoxia-reoxygenation injury. Cultured bovine aorta endothelial cells were placed in an anoxic chamber (0.5% O(2), 5% CO(2)) for 60 minutes followed by a 90-minute reoxygenation. Collagen (2 microg/mL)-induced platelet aggregation (10(8) platelets/mL), as determined by impedance aggregometry, was potently inhibited in the presence of control endothelial cells (17.4+/-3.3 Omega) at a concentration of 5x10(4) cells/mL, as compared to their absence (68. 2+/-2.2 Omega). Inhibition of platelet aggregation was attenuated in endothelial cells exposed to anoxia-reoxygenation (54.6+/-2.5 Omega). However, preincubation of endothelial cells with transforming growth factor-beta(1) (1.0 ng/mL) for 16 hours partially recovered the inhibitory capability of platelet aggregation by injured endothelial cells (40.6+/-3.8 Omega). Cell viability, confirmed by a trypan blue dye exclusion test, was similar (93-96%), including control, 1.0 ng/mL transforming growth factor-beta(1)- and/or anoxia-reoxygenation-pretreated cells. The capability of platelet inhibition was restored when the endothelial cells were preincubated for 4 hours or more. Restoration of antiplatelet capacity in endothelial cells by transforming growth factor-beta(1) was suggested to be due to several mechanisms, including an increase in nitric oxide synthase activity, up-regulation of prostacyclin release, and restoration of adenosine triphosphate diphosphohydrolase activity, which was attenuated by anoxia-reoxygenation pretreatment. In summary, transforming growth factor-beta(1) released from activated platelets may play a compensatory role in the preservation of endothelial functions to inhibit platelet activation.     

手针与电针对于缺血性脑卒中偏瘫患者的治疗效果

目的 比较手针与电针对于缺血性脑卒中偏瘫患者的治疗效果。方法 选取本院针灸科收治的200例缺血性脑卒中偏瘫患者作为研究对象,按照治疗方法分为手针组与电针组,各100例; 在药物治疗的基础上手针组患者给予手针治疗,电针组患者给予电针治疗; 比较手针组与电针组在治疗中各项观察指标的变化,评估其疗效和安全性。结果 治疗结束后手针组与电针组NIHSS评分均下降,Fugl-Meyer运动评分均升高,Barthel指数均升高,但电针组更加显著(P<0.05); 电针组疗效优于手针组(P<0.05),均未发生不良事件。结论 手针和电针对于缺血性脑卒中偏瘫患者均具有一定康复效应,但电针疗效更加显著。     

香港远志提取物对局灶性脑缺血大鼠脑保护作用与作用机制

目的探讨香港远志提取物对局灶性脑缺血大鼠脑保护作用与机制。方法线栓法制备SD大鼠大脑中动脉闭塞(MCAO)模型,随机分4组:假手术组、模型组及治疗A、B组。治疗组按设定方式给药,假手术组、模型组给1%吐温溶液。观察术后24h神经功能缺损评分(NBDS)、脑梗死体积(IV)、血清神经元烯醇化酶(NSE)及神经元凋亡、Bcl-2、Bax表达。结果与假手术组比较,模型组和治疗组NBDS、IV、NSE、神经元凋亡及Bcl-2、Bax显著增高、Bcl-2/Bax显著降低(P<0.05)。与模型组比较,治疗组NBDS、IV、NSE及神经元凋亡、Bax显著降低、Bcl-2、Bcl-2/Bax显著增高(P<0.05),治疗组间数据亦有显著差异(P<0.05)。结论香港远志提取物预处理,对局灶性脑缺血大鼠脑神经有保护作用,机制可能是促进Bcl-2、抑制Bax表达,上调Bcl-2/Bax比值,阻止神经元凋亡。     

Fibrogenic cytokines and extent of fibrosis in muscle of dogs with X-linked golden retriever muscular dystrophy

Fibrosis in Duchenne muscular dystrophy patients' muscle seems to be mediated by fibrogenic cytokines, particularly transforming growth factor-beta1. Golden retriever muscular dystrophy is a model of Duchenne dystrophy characterised by severe myopathy and muscle fibrosis. We evaluated mRNA levels and protein distribution of transforming growth factor-beta1, connective tissue growth factor and collagens in muscle of golden retriever muscular dystrophy dogs at different ages. Fibrosis occurs early in golden retriever muscular dystrophy dogs and transforming growth factor-beta1 levels tend to be high up to 60 days of age (P=0.019 at 30 days), suggesting that transforming growth factor-beta1 is involved in the early stages of fibrosis. We also found greater expression of connective tissue growth factor in golden retriever muscular dystrophy than control muscle (P=0.0065 at 30 days), suggesting involvement of this molecule in fibrosis progression. Our findings sustain the hypothesis that cytokines are actively involved in fibrosis in golden retriever muscular dystrophy, as it seems to be in humans, and confirm the utility of this model for investigating new therapeutic approaches for Duchenne dystrophy.     

Altered protein markers related to neural plasticity and motor function following electro-acupuncture treatment in rat model of ischemic-hypoxic brain injury

BACKGROUND: Previous studies have demonstrated that acupuncture treatment could ameliorate impaired motor function, and these positive effects might be due to neural plasticity. OBJECTIVE: Myelin basic protein (MBP), microtubule-associated protein 2 (MAP2), growth-associated protein-43 (GAP-43), and synaptophysin (SYN) were selected as markers of neural remodeling, and expression of these markers was evaluated with regard to altered motor function following brain injury and acupuncture treatment. DESIGN, TIME AND SETTING: A completely randomized experiment was performed at the Central Laboratory of Peking University First Hospital from November 2006 to May 2007. MATERIALS: Twenty-four Sprague Dawley rat pups, aged 7 days, were selected for the present experiment. The left common carotid artery was ligated to establish a rat model of ischemic-hypoxic brain injury.METHODS: All animals were randomly divided into three groups: sham operation, model, and electro-acupuncture treatment, with 8 rats in each group. Rats in the model and electro-acupuncture treatment group underwent establishment of ischemic-hypoxic brain injury. Upon model established, rats underwent hypobaric oxygen intervention for 24 hours. Only the left common carotid artery was exposed in rats of the sham operation group, without model establishment or oxygen intervention. The rats in the electro-acupuncture treatment group were treated with electro-acupuncture. One acupuncture needle electrode was inserted into the subcutaneous layer at the Baihui and Dazhui acupoint. The stimulation condition of the electro-acupuncture simulator was set to an amplitude-modulated wave of 0-100% and alternative frequency of 100 cycles/second, as well as frequency-modulated wave of 2-100 Hz and an alternative frequency of 3 cycles/second. Maximal current through the two dectrodes was limited to 3-5 mA. The stimulation lasted for 30 minutes per day for 2 weeks. Rats in the sham operation and model groups were not treated with electro-acupuncture, but only fixed to the table for the same time period. MAIN OUTCOME MEASURES: After 2 weeks stimulation, expression of MBP, MAP2, GAP-43, and SYN were detected in the brain by immunohistochemistry. Motor function was evaluated in the three groups. RESULTS: In the sham operation group, MBP was abundant in the myelinated nerve fibers. In the electro-acupuncture treatment group, however, the corpus callosum exhibited more MBP staining than the model group. MAP2 expression was increased in the model group, and increased further in the electro-acupuncture treatment group compared with the sham operation group. GAP-43 expression in the cerebral cortex was less in model group than in sham operation, but present in abundance in the electro-acupuncture treatment group. SYN expression in the cerebral cortex was less in the model and electro-acupuncture treatment group compared with the sham operation group. There was no significant difference in SYN expression and distribution between the model and electro-acupuncture treatment groups. Motor function of rats in the electro-aeapuncture treatment group was significantly better than the model group (P < 0.05), although function remained lower than the sham operation group (P < 0.05). CONCLUSION: Two weeks of electro-acupuncture treatment improved motor function in rats, and protein markers related to neural plasticity also changed, which may be a mechanism for improved motor function in rats with ischemic-hypoxic brain injury.     

Expression of Bax and Bcl-2 proteins during hypoxia in cerebral cortical neuronal nuclei of newborn piglets: effect of administration of magnesium sulfate

This study tests the hypothesis that administration of magnesium sulfate, an antagonist of the NMDA receptor ion-channel, will prevent the hypoxia-induced alteration in the expression and the ratio of Bax and Bcl-2 proteins in cerebral cortical neuronal nuclear membranes. Anesthetized, ventilated and instrumented newborn piglets were divided into three groups: normoxic controls (Nx), untreated hypoxic (Hx), and magnesium sulfate-treated hypoxic (Mg-Hx) groups. Cerebral hypoxia was induced by lowering the FiO2 (0.05-0.07) for 1 h and the cerebral cortex was harvested immediately for isolation of neuronal nuclei and hypoxia was confirmed biochemically by a decrease in the tissue levels of ATP and phosphocreatine (PCr). Brain tissue PCr (micromol/g brain) was 2.74+/-0.77 (Nx), 0.38+/-0.09 (Hx, P<0.05 vs. Nx) and 0.69+/-0.60 (Mg-Hx, P<0.05 vs. Nx). The density of immunoblotted proteins was expressed as absorbance (Axmm(2)). The expression of Bax protein (Axmm(2)) was 222+/-31 (Nx), 279+/-32 (Hx), and 148+/-44 (Mg-Hx, P<0.05 vs. Hx). Bcl-2 protein expression was 77+/-1.0 (Nx), 37+/-5.0 (Hx) and 46+/-15 (Mg-Hx, P<0.05 vs. Nx). The ratio of Bax to Bcl-2 proteins increased more than twofold during hypoxia as compared to normoxia (7:1 Hx vs. 3:1 Nx). However, in the magnesium sulfate-treated group the Bax:Bcl-2 ratio was similar to normoxic controls. The data demonstrate that magnesium sulfate treatment prevents both the hypoxia-induced increase in Bax protein expression and the alteration of Bax:Bcl-2 protein ratios. We suggest that magnesium sulfate treatment before and during hypoxia may decrease hypoxia-induced programmed cell death by maintaining the normal ratio of Bax to Bcl-2 proteins.     

3-硝基丙酸预处理对大鼠局灶性脑缺血Bcl-2和Bax mRNA表达的影响

目的探讨3-硝基丙酸(3-NPA)预处理对大鼠局灶性脑缺血半暗带Bc l-2和Bax mRNA表达的影响。方法将大鼠腹腔注射3-NPA 20 mg/kg,3 d后制作局灶性脑缺血再灌注模型;采用逆转录聚合酶链反应,观察3-NPA预处理对脑缺血再灌注1 h、6 h、12 h、24 h及48 h额顶部皮质Bc l-2和Bax mRNA表达的影响,并与假手术组和缺血再灌注组比较。结果与假手术组比较,缺血再灌注组和3-NPA预处理组各时间点Bc l-2和Bax mRNA表达极显著增强(均P<0.01);与缺血再灌注组比较,3-NPA预处理组各时间点Bc l-2mRNA表达显著增强(均P<0.05),再灌注12~48 h Bax mRNA的表达显著降低(均P<0.05)。结论增强Bc l-2的表达、抑制Bax的表达,可能是3-NPA预处理抑制细胞凋亡、诱导脑缺血耐受的机制之一。     

针刺及艾灸足三里穴缓解大鼠运动疲劳作用的比较

背景：已有诸多实验证实针灸足三里穴能够有效地缓解运动疲劳。 目的：观察毫针针刺及艾灸足三里穴(ST36)对运动疲劳大鼠的运动耐力、骨骼肌微循环及抗氧化酶活性的影响,探讨针刺及艾灸两种不同疗法缓解运动疲劳作用的差异。 设计、时间及地点：随机动物实验,于2008-06/07在华南师范大学光子中医学实验室完成。 材料：SPF级雄性成年SD大鼠24只,体质量220~260 g。 方法：SD大鼠24只,适应性游泳后被随机分为正常对照组、模型组、艾灸组及针刺组,每组6只。采用无负重游泳方式建立大鼠运动疲劳模型,艾灸组及针刺组在游泳运动的同时,分别采用毫针及艾灸刺激足三里穴,1次/d,共10 d。末次力竭运动结束后检测指标。 主要观察指标：大鼠骨骼肌微循环及线粒体内超氧化物歧化酶、谷胱甘肽过氧化酶的活性。 结果：实验第9天,艾灸组大鼠的运动耐力显著高于同时间点模型组的运动耐力(P < 0.05);实验第11天,艾灸组和针刺组的大鼠的运动耐力均显著高于模型组的大鼠同时间点的运动耐力(P < 0.05),艾灸组和针刺组相比差异无显著性意义 (P > 0.05)。艾灸组双侧胫骨前肌的血流灌注量均显著高于模型组(P < 0.05);针刺组腹直肌的血流灌注量显著高于模型组(P < 0.05)。艾灸组双侧胫骨前肌线粒体内的超氧化物歧化酶、谷胱甘肽过氧化物酶活性显著高于模型组(P < 0.05); 针刺组双侧胫骨前肌线粒体内的超氧化物歧化酶、谷胱甘肽过氧化物酶活性显著低于艾灸组(P < 0.05)。 结论：艾灸足三里穴能够有效地提高运动疲劳大鼠骨骼肌线粒体抗氧化酶活性、增加骨骼肌血流灌注,缓解外周骨骼肌的运动疲劳,提高运动耐力,其效应优于针刺足三里穴。     

Cytokines involved in CNS manifestations caused by Mycoplasma pneumoniae

Mycoplasma pneumoniae sometimes causes central nervous system manifestations, which may involve the host immune response, as the organism does not directly damage neural cells, or release toxins. Therefore we measured the levels of interleukin-6, interleukin-8, interleukin-18, interferon-gamma, tumor necrosis factor-alpha, and transforming growth factor-beta1 in serum and cerebrospinal fluid samples from patients who manifested central nervous system manifestations during acute M. pneumoniae infection. The subjects were nine patients with early-onset encephalitis (central nervous system disease onset within 7 days from the onset of fever), four with late-onset encephalitis (onset at 8 days or later), three with encephalitis but without fever, and three with aseptic meningitis. Intrathecal elevations of interleukin-6 and interleukin-8 in all four types of central nervous system manifestations, and of interleukin-18 in late-onset encephalitis were observed. None of the cerebrospinal fluid samples contained detectable levels of interferon-gamma, tumor necrosis factor-alpha, or transforming growth factor-beta1. In conclusion, interleukin-6, interleukin-8, and interleukin-18 might be involved in the inflammatory process leading to the central nervous system manifestations caused by M. pneumoniae.     

Atypical inflammation in the central nervous system in prion disease

The inflammatory response in prion diseases is dominated by microglial activation. Contrary to their profile in vitro none of the pro-inflammatory cytokines interleukin-1beta, interleukin-6, or tumour necrosis factor-alpha are significantly upregulated in the ME7 model of prion disease. However, two major inflammatory mediators are elevated: transforming growth factor-beta1 and prostaglandin E2. This cytokine profile is the same as that reported for macrophages during phagocytosis of apoptotic cells and indeed transforming growth factor-beta1 and prostaglandin E2 are responsible for the downregulated phenotype of these macrophages. Transforming growth factor-beta1 may also have roles in extracellular matrix deposition and in amyloidogenesis and may play a direct role in disease pathogenesis. There is also now evidence to suggest that a peripheral infection, and its consequent systemic cytokine expression, may drive central nervous system cytokine expression and perhaps exacerbate disease.     

胃扩张和针刺大鼠四白传入信息对孤束核神经元放电整合的影响

目的观察胃扩张和针刺大鼠凹白传入信息对孤束核(NTS)神经元放电整合的影响。方法采用细胞外记录方法,先找到胃扩张相关神经元;以足阳明胃经凹白穴为刺激点作为研究对象,并以同名经的地仓和内庭、手太阳小肠经的颧髎、非经非穴四白旁开点为对照,在刺激点上用手针施以捻转手法刺激30s,在NTS寻找对来自体表刺激点有反应的胃扩张相关神经元。结果52只大鼠共记录到104个NTS神经元放电,104个NTS神经元中与胃扩张相关的有47个。47个胃扩张相关神经元中兴奋性神经元32个,抑制性神经元15个。47个NTS胃扩张相关神经元中对来自体表刺激有反应的神经元出现比例,在针刺凹白、地仓、内庭、颧髎、凹白旁开点后分别为80.85%、61.70%、57.45%、38.30%、36.17%。统计表明,在胃扩张相关神经元中,针刺凹白后NTS有反直神经元出现的几率,极显著高于针刺地仓、内庭、颧髎、四白旁开点后有反应神经元出现的几率(P<0.01);针剌地仓和内庭后NTS有反直神经元出现的几率,亦均极显著高于刺激颧髎和凹白旁开点的有反应神经元出现几率(P<0.01)。结论　与非经非穴点以及手太阳小肠经穴比较,NTS中胃扩张相关的神经元与足阳明冒经关系更密切;足阳明经中远近腧穴的信息传入在NTS亦存在着一定的差异,这提示腧穴和经脉均有一定的特异性。     

电针干预自发性高血压大鼠主动脉丝裂原活化蛋白激酶磷酸酶1及磷酸化细胞外信号调节激酶1/2蛋白的表达

背景：近年来大量临床研究表明针刺风池、太冲、曲池等穴位能有效降低血压,可用于高血压,但对其治疗的分子机制尚未阐明。 目的：观察针刺大鼠风池、太冲、曲池等穴位对丝裂原活化蛋白激酶信号转导调控系统的影响,从而探讨针刺治疗高血压的分子机制。 方法：选取8月龄自发性高血压雄性Wistar大鼠14只,随机分为针刺组和模型组,每组7只;另选取同月龄正常血压雄性Wistar-Kyoto大鼠7只作为对照组。对针刺组大鼠采用电针针刺双侧风池、曲池和三阴交穴,毫针刺太溪和太冲穴。3周后采用RT-PCR方法检测各组大鼠主动脉组织丝裂原活化蛋白激酶磷酸酶1 mRAN的表达,Western blot方法检测丝裂原活化蛋白激酶磷酸酶1、磷酸化细胞外信号调节激酶1/2蛋白表达。 结果与结论：与对照组比较,模型组主动脉组织磷酸化细胞外信号调节激酶1/2蛋白表达水平升高,丝裂原活化蛋白激酶磷酸酶1 mRNA及其蛋白表达水平降低(P < 0.01);与模型组比较,针刺组主动脉组织磷酸化细胞外信号调节激酶1/2蛋白表达水平降低,丝裂原活化蛋白激酶磷酸酶1 mRNA及其蛋白表达水平升高(P < 0.05)。提示针刺治疗自发性高血压大鼠可能是通过调控丝裂原活化蛋白激酶信号转导途径,增强磷酸化细胞外信号调节激酶1/2蛋白表达,降低丝裂原活化蛋白激酶磷酸酶1蛋白表达,从而改善血管重塑,降低血压。