Objective To study the effects of cyanide compounds intoxication and hemorrhagic shock on pulmonary gas exchange and phospholipids contents of the rats.Methods SD rats were randomized equally into sham operation group (SO),hemorrhagic shock group (HS),potassium cyanide intoxication group (KCN)and hemorrhagic shock combined potassium cyanide intoxication group(HS+KCN).Blood samples used for blood-gas analysis were taken at the time point of 15min,30min,60min and 120min after cessation of exposure,Evan’s blue (EB)in lung tissues was examined by spectrophotometric method and the content of phospholipids was assayed with a biochemistry method.Results Compared with KCN group,PAaDO2 and RI were obviously increased in (HS+KCN)group.Combined hemorrhagic shock and KCN intoxication produced a significant increasing effect on EB content and decreasing effect on phospholipids content.Conclusions Hemorrhagic shock and cyanide intoxication may induce a combined inj ury effect on rats lung.

Objective To investigate effects of hemorrhagic shock complicated with cyanide poisoning on heart function of rat.Methods SD rats were randomly and equally divided into sham operation group (SO), hemorrhagic shock group (HS ), potassium cyanide poisoning group (KCN ) and hemorrhagic shock complicated with potassium cyanide poisoning group (HS+KCN).The activities of SOD,GSH-Px and the content of MDA were assayed in rat myocardium mitochondria.The left ventricular systolic pressure (mLVSP),+dp/dtmax and heart rate (HR)were recorded by physiological function signal processing system. The arterial blood was taken for myocardium zymogram detection (AST,LDH,CK,and CK-MB ). Results Compared with KCN group,SOD,GSH-Px activities and MDA content obviously decreased,the mLVSP,+ dp/dtmax and HR changed obviously in the rats of HS + KCH group, meanwhile, the myocardium zymogram of rats showed significant changes accordingly.Conclusions Under the condition of hemorrhagic shock,complicated cyanide poisoning would impair the cardiac function more severely,further study on effective measures for prevention and treatment is urgently required.

OBJECTIVE To investigate effects of cyanide compounds on cerebral oxygen and energy metabolism in rats' brain under hemorrhagic shock.METHODS SD rats were randomizedly divided equally into sham operation group (SO),hemorrhagic shock group (HS),potassium cyanide intoxication group (KCN) and hemorrhagic shock combined potassium cyanide intoxication group (HS+KCN).Blood samples were taken from internal jugular bulb to determine SjvO2.The contents of ATP,ADP and AMP in rats' cortex were assayed by high performance liquid chromatography (HPLC).The activities of Na+-K+-ATPase and Ca2+-ATPase were measured by usingwith ATPase detect kit.RESULTS Compared with KCN group,SjvO2obviously decreased in (HS+KCN) group,ATP contents significantly decreased,but ADP and AMP content increased.The activities of Na+-K+-ATPase and Ca2+-ATPase in rats' cortex were obviously inhibited.CONCLUSION Hemorrhagic shock could markedly disturb cerebral oxygen and energy metabolism in rats' brain intoxicated by KCN.

Objective To explore the role of glutamate transporters in the neurotoxicity induced by rotenone.Methods Astrocytes isolated from newborn rats,were cocultured with PC12 cells,then were divided into 6 groups:control group,rotenone treated group,DHK pretreated group(Ⅰ and Ⅱ),and PDC pretreated group(Ⅰ and Ⅱ).Extracellular glutamate concentrations were detected by high performance liquid chromatography(HPLC),and the uptake ability of glutamate was determined with isotope labeling method.Results It was showed that the glutamate uptake ability in astrocytes pretreated with PDC was significantly decreased compared with rotenone treated group,while those in astrocytes pretreated with DHK failed to show any significant change.Conclusion GLAST rather than GLT-1 may play a crucial role in excitotoxicity induced by rotenone.

Objective To explore the effect of rotenone on the levels of excitatory amino acids(EAAs) and the expression of NMDA receptor(NMDAR) in the striatum of rat.Methods Chronic rotenone intoxicated model was established through subcutaneously implanting a mini-osmotic pump with rotenone in the back of rat.The levels of glutamate(Glu) and aspartic acid(Asp) in striatum of rat were detected by high performance liquid chromatography(HPLC),and the expression of NMDAR subunits' mRNA,such as NR1,NR2A and NR2B,were measured by RT-PCR.Results Compared with those in control group,the levels of Glu and Asp in striatum of rotenone treated groups were increased significantly(P 0.01),and mRNA expressions of NR1 and NR2A in all rotenone treated groups were obviously up-regulated,and the expression of NR2B mRNA in high dose(4mg/kg) rotenone treated group was also enhanced significantly.Conclusion It was suggested that rotenone could induce the increase of EAAs in the striatum and as a consequence,the expression of NMDAR mRNA was also up-regulated,which might be the possible mechanism of rotenone neurotoxicity.

Objective To evaluate the effects of anti-arrhythmic peptide 10(AAP-10)and activator of gap junction on metabolism of glutamate in rotenone exposed astrocytes.Methods MTT assay was used to evaluate the toxicity of rotenone on astrocytes,the level of extracellular glutamate was determined by HPLC,the uptake of glutamate was detected with labeled isotope,and the expression of GLAST mRNA was observed by RT-PCR.Results Compared with 1.0 μmol/L rotenone intoxication group,the viabilities of cell culture were significantly increased and the levels of extracellular glutamate were obviously decreased.The function of glutamate uptake and the expression of GLAST mRNA were enhanced in AAP-10 group(250 nmol/L).Conclusions Enhancement of gap junction could significantly ameliorate the disorder of glutamate metabolism in astrocytes induced by rotenone,which might be related to the change of GLAST.

OBJECTIVE To observe some of bio-chemical indice of rats striatum poisoned by rotenone. METHODS Apply the method of back-implantation mini-effusion pump to observe influence of different concentration rotenone on the rat striatum. Utilize Fluoro-Jade B combined Fluorescent dying to observe the change conditions of the neural cells of the poisoned rat striatum. HPLC is used to measure contents of ATP, ADP, and AMP in striatum. Apply bio-chemical lab to analyze the activities of Na+ -K+ -ATPase and Ca2+ -ATPase. RESULTS There appeared a large quantity of positive degenerative neurons in the poisoned rats striatum, but the solution control group did not show similar change. Compared with the solution control group, the ATP contents in the rat striatum of the 2.0 mg/kg and 4.0 mg/kg rotenone groups were significantly decreased (P < 0.01), while the contents of ADP and AMP were relatively raised. With the increase of the poison contents, the activities of Na+ -K -ATPase and Ca2+ -ATPase were inhibited to some degrees. The difference has statistical significance. CONCLUSION Rotenone could cause the decrease of the ATP content and inhibit activities of Na+ -K+ -ATPase and Ca2+ -ATPase.

目的观察激活的星形胶质细胞条件培养液(ACM)在鱼藤酮所致PC12细胞氧化损伤过程中的作用。方法收集激活的ACM,加入到鱼藤酮染毒PC12细胞中,观察其对染毒神经元胞内丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性的影响,并检测细胞活性氧(ROS)水平和线粒体呼吸链复合物Ⅰ的活性变化。结果ACM可明显降低染毒PC12细胞MDA和ROS水平;与完全培养基处理比较,ACM能有效增加染毒神经元SOD和GSH-Px的合成和释放,保护和提高抗氧化酶活性。结论ACM能有效抑制鱼藤酮诱导的PC12细胞氧化损伤。

目的观察星形胶质细胞条件培养液(ACM)对鱼藤酮染毒嗜铬细胞瘤细胞(PC12)的神经保护作用。方法将新生SD大鼠中脑星形胶质细胞分离、纯化并培养至第三代后,收集星形胶质细胞条件培养液加入到鱼藤酮染毒PC12细胞中,观察其对染毒神经元形态、活力及其合成与释放一氧化氮(NO)、乳酸脱氢酶(LDH)和胞膜ATP酶(ATPase)的影响。结果20%ACM可明显改善染毒神经元的形态和活力;与对照组比较,20%ACM还有效减少了染毒神经元NO、LDH的合成和释放,保护和提高了ATPase活性。结论ACM明显促进鱼藤酮染毒PC12细胞的活力,其机制可能与减少NO、LDH合成释放及保护细胞膜ATPase的活性有关。

[Objective] To investigate the intracellular calcium concentration alteration induced by rotenone and the mechanism of ion flux in astrocytes.[Methods] Rat mesencephal astrocytes were cultured in vitro.Intracellular calcium concentrations were measured by laser scanning confocal microscope and Fluo-4 fluorescence dye technique.[Results] Rotenone significantly increased intracellular calcium concentration in a dose-dependent manner.Intracellular calcium store released blocker(TMB-8),Ca2+-free bath,and calcium channel blocker(MK-801 and nimodipine),which inhibited the elevation of intracellular calcium concentration induced by rotenone,but the inhibiting time and level were also significantly different.[Conclusion] Intracellular Ca2+ elevation induced by rotenone is due to both the release of intracellular calcium store and extracellular calcium entry,but the release of intracellular calcium stores occurs in the latter phase.Rotenone-induced extracellular calcium entry is mainly through voltage-operated Ca2+ channel.

Objective To study the effects of rotenone on intercellular gap junction function of astrocytes.Methods Astrocytes isolated from newborn rats were cocultured with rotenone and a common cell strain-PC 12 cells;the intercellular gap junction function of rotenone-exposed astrocytes was observed by scrape-loading and dye transfer technique(SLDT),and the expression of CX43 mRNA and protein were detected by RT-PCR and Western Blot.Results The results showed that the the intercellular gap junction function of astrocytes got obviously inhibited by rotenone with a dose-dependent manner;and the expressions of CX43 mRNA and protein were also decreased significantly in 0.5 and 1.0 μmol/L rotenone exposed group(P0.01).Conclusions The results suggested that rotenone could reduce the express of CX43 and inhibite the intercellular gap junction function of astrocytes,which might be one of the important mechanisms of the neurotoxicity of rotenone.

Objective To study the effects of rotenone on glutamate uptaking in primary cultured astrocytes.Methods The extracellular glutamate was detected by high performance liquid chromatography(HPLC),the uptaking ability of glutamate was detected with isotope lebeling method and the mRNA and protein expression of glutamate transporter were detected using RTPCR and Western blot,respectively.Results It was showed that the extracellular rotenone level was obviously risen,shown the uptaking function of astrocyte to glutamate was decreased,Meanwhile,the expression of GLAST(glutamate/aspartate transporter)mRNA and protein also decreased significantly in 1.0 and 2.0 μmol/L rotenone exposed groups,but the expression of glutamate transporter-1(GLT-1)enhanced obviously.Conclusions It was suggested that rotenone might reduce the uptaking ability of astrocyte to glutamate,induce the extracellular level of Glu risen,the down-regulation of GLAST may be the main reason of elevated extracellular Glu level induced by rotenone,while the increased expression of GLT-1 might represent a self-protective mechanism by nerve cells for limiting the neurotoxicity by Glu.

[Objective] To study the effects of rotenone on glutamate-glutamine circulation in rats striatum.[Methods] 24 Sprague-Dawley rats were divided into four groups randomly:the control group,solvent control group,2.0 mg/kg and 4.0 mg/kg rotenone treated groups.The concentration of glutamate and glutamine in rats striatum were assayed by high performance liquid chromatography(HPLC).The expression of glutamine synthetase(GS)mRNA was measured by RT-PCR,and the activities of GS and phosphate activated glutaminase(PAG)were determined by using GS and PAG detect kit.[Results] Compared with control group,the release of glutamate in rats striatum increased(P 0.01)and the concentration of glutamine decreased(P 0.01),the activity of GS declined,but PAG activity elevated obviously in 4.0 mg/kg rotenone treated group.[Conclusion] Glutamate-glutamine circulation was damaged in the brain of rotenone treated rats,which may be responsible for neuron degeneration induced by rotenone.

轴突、少突胶质细胞及其形成的髓鞘是构成中枢神经系统白质的重要成分。从功能解剖角度来看,脑白质主要起连接作用,将皮层与灰质连接起来,构成完整的功能体系。白质的完整是保证轴突传导功能的基础,脑白质变性损伤是神经功能障碍的重要原因之一。本文从兴奋性毒作用的角度,综述了脑白质变性损伤机制的研究进展。

蛋白酶激活受体-2是一种细胞膜表面受体,属于G蛋白偶联受体家族,广泛分布于全身多种组织器官,激活后参与许多疾病的发生和发展过程,尤其在中枢神经系统中所起的作用受到人们越来越多的关注。

半乳凝素-1(galectin-1)是半乳凝素家族成员之一,具有高度保守性,参与细胞生长、黏附、迁移以及肿瘤发生发展等过程。近年来研究还发现半乳凝素-1可能具有神经保护作用。

Objective To explore the toxic effect of rotenone on striatum and expression of connexin 43(CX43)in rats. Methods SD rats were administrated with rotenone via Alzet osmotic pump for 28 days;morphological changes of neurons and astrocytes in striatum were observed by light microscope with HE staining and immunohistochemistry method;while the expression of CX43 mRNA was detected by RT-PCR and the amount of CX43 protein was measured by Western blot. Results The results showed that the autonomic activities were decreased in rotenone exposed rats,quite similar to the manifestation of Parkinson's syndrome.The morphological changes of neurons and astrocytes in striatum were quite obvious,and the CX43 mRNA expression and CX43 protein amount were also increased significantly in rats treated with 2.0 mg/kg rotenone(P 0.05),however,which were obviously decreased in rats treated with 4.0 mg/kg rotenone(P 0.05). Conclusions It is suggested that rotenone may impair neurons in striatum of mesencephalon in rats and induce Parkinsonism-like symptoms,which might be attributed to the abnormal expression of CX43.

Objective To study the toxic effects of rotenone on the activity of CaMKⅡ in primary cultured astrocytes.Methods MTT assay and morphological observation were used to evaluate the toxicity of rotenone on astrocytes,intracellular calcium concentrations were measured by confocal laser scanning microscopy and Fluo-4 fluorescence dyeing techniques,the expression of CaMKⅡα and CaMKⅡ β mRNA were detected by RT-PCR,and the activity of phosphory-CaMKⅡ protein was measured by Western blot.Results The viability of cultural cells was significantly decreased after treated with 1.0 and 2.0 μmol/L rotenone,rotenone was showed to increase intracellular calcium concentration in a dose-dependent manner,the expression of CaMKⅡα mRNA decreased significantly in 1.0 and 2.0 μmol/L rotenone intoxication groups(P<0.05),but the expression of phosphory-CaMKⅡ enhanced obviously(P<0.01).Conclusions The elevation of astrocytes[Ca2+]i and the inhibition of CaMKⅡ activity induced by rotenone may be the main reason of its neurotoxicity on CNS.

MAPK/ERK信号通路是目前研究最为活跃的信号转导通路之一,很多研究表明该通路具有调控细胞增殖分化、细胞恶性转化以及学习记忆等多种功能.本文简要综述了MAPK/ERK信号转导通路与学习记忆功能的研究进展.

长链脂肪醇(Policosanol)是指碳链长度为26～32个碳原子的醇类物质.本实验所用的长链脂肪醇经气相色谱分析含二十八烷醇16.91%、三十烷醇56.14%、三十二烷醇11.56%、其它物质为15.39%[1].左旋肉碱(又称肉毒碱,L-carnitine,以下简称肉碱),化学名L-3-羟-4-三甲基铵丁酸,是存在于人体内的一种类维生素,其主要生理功能是将长链脂肪酸转运入线粒体内氧化,为人体肌肉细胞及心肌细胞提供能量.是人体内不可缺少的营养物质[2].本实验将2种物质合用于小鼠的抗疲劳实验,探索其合用的抗疲劳效果.