美满霉素对阿尔茨海默病大鼠认知损伤及海马 BDNF、 Bcl-2 和 Bax 表达的影响

目的 观察美满霉素对阿尔茨海默病（AD）大鼠认知功能和海马脑源性神经营养因子（BDNF）、凋亡相关因子 Bcl-2 和 Bax 表达的影响, 探讨美满霉素对 AD 大鼠脑保护作用的机制。 方法 侧脑室注射 Aβ25-35 建立 AD 大鼠模型。 30 只健康雄性 SD 大鼠随机分成对照组、模型组和治疗组, 每组 10 只。 对照组和模型组腹腔内注射生理盐水 1 mL/（kg· d）, 治疗组腹腔注射美满霉素 50 mg/（kg· d）, 均持续 14 d。 Morris 水迷宫检测行为学变化, 蛋白免疫印迹（Western blotting）法和酶联免疫吸附试验（ELISA）法检测海马 BDNF、Bcl-2 和 Bax 蛋白的表达, 原位末端标记（TUNEL）法检测海马神经元凋亡率。 结果 美满霉素可以明显提高 AD 大鼠学习记忆能力, 上调 AD 大鼠海马 BDNF、Bcl-2 表达, 下调 Bax 表达, 减少海马神经元凋亡。 结论 美满美素可以通过促进神经元的生长、抑制神经元的凋亡发挥脑保护作用。     

硫酸茯苓多糖对抑郁症大鼠海马AMPA受体表达的影响

目的探讨硫酸茯苓多糖(SP)是否通过调节α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体的表达而生产抗抑郁作用。方法实验设假手术组、抑郁症模型组、SP(25,50和100 mg·kg~(-1))剂量组及SP(100 mg·kg~(-1))+GYKI 52466(3 mg·kg~(-1))剂量组,每组12只。大鼠卵巢摘除加慢性不可预知性温和应激(CUMS)法诱导抑郁症动物模型。连续灌胃和腹腔注射给药21 d。糖水消耗、强迫游泳及敞箱试验观察大鼠行为变化。尼氏染色观察大鼠海马神经元病理变化。免疫组化法观察海马脑源性神经营养因子(BDNF)的表达。Western蛋白印迹法检测海马AMPA受体GluR1、磷酸化GluR1及c AMP反应元件结合蛋白(CREB)、p-CREB和BDNF蛋白表达。结果与模型组相比,SP各给药组大鼠强迫游泳不动时间明显降低而自发活动明显增加,同时糖水摄入量及糖水偏爱百分比明显升高(P<0.05,P<0.01),海马神经元损伤显著减轻,GluR1和GluR1磷酸化水平及p-CREB和BDNF表达均明显升高。但与SP(100 mg·kg~(-1))剂量组相比,SP+GYKI 52466组大鼠强迫游泳不动时间又明显增加而自发活动又明显减少,糖水摄入量及糖水偏爱百分比又明显降低,海马GluR1和p-GluR1及p-CREB和BDNF表达又明显降低(P<0.01)。结论 SP具有抗抑郁作用,其机制可能通过增强海马GluR1受体功能,进而上调海马p-CREB和BDNF蛋白表达有关。     

丙泊酚麻醉对新生期大鼠海马BDNF及Trk-B的影响

目的观察新生期大鼠丙泊酚麻醉对空间学习记忆能力发育的影响。方法 7日龄SD大鼠45只,随机平均分为3组:空白对照组,不做任何处理;丙泊酚麻醉组,腹腔注射丙泊酚25 mg.kg-1,每20 min给予首次剂量的1/2,麻醉维持2 h;中长链脂肪乳注射组,腹腔注射脂肪乳25 mg.kg-1,每隔20 min给予首次剂量的1/2。待麻醉后4 h,每组随机处死6只,免疫组化法分析海马的caspase-3表达,蛋白免疫印迹法测定BDNF和Trk-B的水平。其余同窝饲养至40 d时行Morris水迷宫测定。结果与对照组比较,丙泊酚麻醉组海马BDNF及Trk-B表达明显减少、caspase-3表达明显增加(P<0.05),脂肪乳组与对照组差异无显著性。Morris水迷宫实验中,丙泊酚组大鼠寻找隐藏平台潜伏期较对照组明显延长(P<0.05),在记忆保留实验中丙泊酚组大鼠穿越平台次数明显少于对照组(P<0.05),脂肪乳组与对照组均无明显差异。结论新生期大鼠丙泊酚麻醉可影响空间认知功能发育,该效应可能与新生期海马内神经元凋亡增加,BDNF途径抑制有关。     

电针对阿尔茨海默病大鼠学习记忆及海马基质金属蛋白酶-2,9的影响

目的 探讨电针对阿尔茨海默病大鼠学习记忆及海马基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)的影响.方法 采用链脲霉素(STZ)侧脑室注射建立大鼠阿尔茨海默病模型,将大鼠分为4组,电针组针刺双肾俞、双太溪、双足三里、百会、大椎等穴位治疗.西药组给予安理申混悬液每天0.5 g/kg,正常组和模型组用等量0.9%氯化钠溶液治疗6周.用Morris水迷宫检测大鼠的学习记忆能力,同时免疫组织化学法测定MMP-2、MMP-9表达.结果 与正常组相比,模型组大鼠学习能力降低(P < 0.05).给予电针刺激后,大鼠学习记忆明显改善,与模型组相比差异具有统计学意义(P < 0.05).模型组海马组织MMP-2、MMP-9表达明显增高(22.73±2.61,23.80±2.52),电针组、西药组海马组织MMP-2、MMP-9表达均低于模型组(P < 0.05).结论 电针能改善阿尔茨海默病大鼠学习记忆力功能,可能与抑制细胞凋亡有关.     

注射用鼠神经生长因子对大鼠缺血脑组织神经生长因子及脑源性神经营养因子表达的影响

目的研究注射用鼠神经生长因子对大鼠缺血脑组织中脑源性神经营养因子(BDNF)和神经生长因子(NGF)表达的影响。方法健康雄性Wistar大鼠,采用改良的Zea-Longa法建立大脑中动脉局灶缺血模型(MCAO)。实验分为模型对照组和注射用鼠神经生长因子组,每组各20只大鼠。注射用鼠神经生长因子组予以腹腔内注射鼠神经生长因子,每天1次,每次9 k U;模型对照组每天注射等量0.9%氯化钠注射液。72 h后行神经功能评分,然后处死,分别取脑梗死周围、海马区和梗死区脑组织作免疫组织化学染色检测BDNF、NGF蛋白表达变化,原位杂交检测BDNF m RNA、NGF m RNA的表达。结果治疗3 d后注射用鼠神经生长因子组神经功能缺损评分低于模型对照组(P<0.05)。注射用鼠神经生长因子治疗组BDNF和NGF蛋白、BDNF m RNA和NGF m RNA在梗死区周围和海马区表达均高于模型对照组,差异有统计学意义(P<0.05),在梗死区差异均无统计学意义(P>0.05)。结论注射用鼠神经生长因子可上调大鼠梗死区周围和海马区缺血脑组织中BDNF和NGF的表达,并可能通过此机制起到保护作用。     

七氟醚麻醉对子代大鼠海马神经元自噬的影响

目的探讨七氟醚麻醉对子代大鼠海马神经元自噬的影响。方法孕5~7 d的SD雌鼠18只,采用随机数字表法将其分为3组(n=6):对照组(C组)、七氟醚组(S组)、七氟醚+自噬抑制剂wortmannin组(SW组)。C组母鼠仅置于麻醉箱内,通入相同流量氧气暴露4 h,S组和SW组母鼠给予2.8%七氟醚吸入麻醉4 h。SW组子代大鼠于出生后1~3 d腹腔注射0.5 mg/kg自噬抑制剂wortmannin,C组和S组于相同时点腹腔注射等量的生理盐水对照。于出生后22 d,对各组子代大鼠进行水迷宫实验,测定其学习记忆功能,出生后28 d,完成水迷宫测试后处死子代大鼠,并取其海马组织。采用TUNEL染色测定子代大鼠海马组织内神经元的凋亡率,采用Western blot法检测子代大鼠海马组织内微管相关蛋白l轻链3(LC3)Ⅱ/LC3Ⅰ比值作为自噬活性的指标。结果 S组、C组子代大鼠逃避潜伏期较C组,穿越平台次数比较差异无统计学意义(P>0.05),SW组子代大鼠逃避潜伏期较C组延长,穿越平台次数较C组减少(P<0.05)。与C组比较,S组和SW组子代大鼠海马神经元的凋亡率和LC3Ⅱ/LC3Ⅰ比值均升高(P<0.05);与S组比较,SW组子代大鼠海马神经元的LC3Ⅱ/LC3Ⅰ比值降低(P<0.05)。结论大鼠孕期七氟醚麻醉对子代学习记忆功能无影响,其机制与激活生理性自噬对抗神经元凋亡有关。     

异丙酚对大鼠脑内nNOS表达及NO含量的影响

目的通过观察异丙酚对大鼠海马以及基底前脑内神经型一氧化氮合酶(nNOS)的表达和一氧化氮(NO)释放量的影响,探讨异丙酚的中枢麻醉机制。方法选择正常雄性Wistar大鼠30只随机分为三组,分别腹腔注射异丙酚50mg/kg(P50组)、100mg/kg(P100组)及生理盐水10mL/kg(NS组)。10min后,各组大鼠经主动脉灌注生理盐水,随机选取5只取新鲜鼠脑,分离海马及基底前脑,分光光度法测其NO释放量;每组另5只大鼠继续灌注固定液后取脑,免疫组化检测海马及基底前脑内nNOS的表达。结果与对照组NS组相比,海马及基底前脑内nNOS表达量、NO释放量都显著减少(P<0.05)。结论异丙酚可能通过抑制海马及基底前脑内nNOS表达,使NO的释放减少而发挥麻醉作用。     

CREB磷酸化介导半枝莲黄酮改善BI-D1870所致大鼠记忆障碍

目的：探讨半枝莲黄酮(SBFs)通过促进CREB磷酸化改善BI-D1870所致大鼠记忆障碍。方法：腹腔注射BI-D1870建立大鼠记忆障碍模型,Morris水迷宫筛选记忆障碍成功模型,模型成功大鼠随机分为模型组、SBFs组和CREB磷酸化激活剂Rolipram组,SBFs组大鼠灌胃140 mg·kg^–1 SBFs,Rolipram组大鼠腹腔注射0.5 mg·kg^–1 Rolipram。Morris水迷宫检测大鼠学习记忆能力;苏木精-伊红(HE)染色观察海马皮层神经元形态;免疫组化(IHC)检测海马回与神经再生相关蛋白DCX表达;免疫荧光(IF)检测海马和皮层p-CREB-Ser142蛋白表达;实时荧光定量(qPCR)检测海马和皮层BDNF、TrkB和CREB的mRNA表达;Western blot检测海马和皮层BDNF、TrkB和p-CREB-Ser133蛋白表达。结果：腹腔注射BI-D1870可引起大鼠学习记忆障碍,神经病理学改变,海马回DCX蛋白表达降低,海马和皮层p-CREB-Ser142蛋白表达增加,BDNF、TrkB和CREB的mR...     

伊潘立酮联合阿立哌唑对慢性铝暴露痴呆模型大鼠海马神经元及BDNF和NGF表达的影响

目的旨在探讨伊潘立酮联合阿立哌唑对慢性铝暴露痴呆模型大鼠海马神经元及BDNF和NGF表达的影响。方法选择6周龄清洁级SD雄性大鼠建立慢性铝暴露痴呆模型,分为模型组、伊潘立酮组、阿立哌唑组和联合组,每组各20只,并同时设健康大鼠20只为对照组。伊潘立酮组大鼠接受0.05 g/ml伊潘立酮0.25 g/kg灌胃,阿立哌唑组大鼠接受0.05 g/ml阿立哌唑0.25 g/kg灌胃,联合组大鼠接受0.05 g/ml伊潘立酮0.25 g/kg和0.05 g/ml阿立哌唑0.25 g/kg灌胃,对照组大鼠和模型组大鼠均接受同等剂量生理盐水灌胃。各组大鼠均进行学习记忆测试和旷场实验测试。Annexin V/FITC法测定大鼠海马组织神经元凋亡率、RT-PCR法检测大鼠海马组织BDNF及NGF mRNA表达、Western blot法检测大鼠海马组织BDNF及NGF蛋白表达。结果对照组和联合组大鼠电击次数显著低于模型组大鼠(P0.05),而水平穿越格数和竖立次数显著高于模型组大鼠(P0.05)。联合组大鼠电击次数低于伊潘立酮组和阿立哌唑组大鼠,水平穿越格数和竖立次数高于伊潘立酮组和阿立哌唑组大鼠,但组间差异无统计学意义(P0.05)。对照组、联合组、伊潘立酮组和阿立哌唑组大鼠海马组织神经凋亡率均显著低于模型组大鼠(P0.05)。对照组、联合组、伊潘立酮组和阿立哌唑组大鼠海马组织BDNF及NGF mRNA、蛋白表达均显著高于模型组大鼠(P0.01)。结论伊潘立酮联合阿立哌唑能显著抑制慢性铝暴露痴呆模型大鼠海马神经元凋亡,上调BDNF及NGF表达。     

TNF-α在丙泊酚诱发的神经元凋亡及认知功能障碍中的作用

目的探讨TNF-α在丙泊酚诱发的海马神经元凋亡及远期认知功能障碍中的作用。方法 7 d龄(P7)SD大鼠随机分为3组:Control组,无任何处理;P(single)组,腹腔注射丙泊酚50 mg·kg~(-1);P(repeated)组,腹腔注射丙泊酚50mg·kg~(-1),每天1次,共7次。丙泊酚两组于麻醉结束后2 h(对照组则分别对应上述两时间点)采用Western blot法检测海马组织TNF-α含量。另取P7大鼠随机分为5组:Control组;P(single)组;P(repeated)组;P(single)+ETN组,腹腔注射丙泊酚50 mg·kg~(-1)前30 min侧脑室注射依那西普0.4 mg·kg~(-1);P(repeated)+ETN组,腹腔注射丙泊酚50 mg·kg~(-1),每天1次,共7次,第1次和第4次丙泊酚注射前30min侧脑室注射依那西普0.4 mg·kg~(-1)。于P7、P13、P21、P35时间点,各组采用免疫组化法检测海马CA1区神经元凋亡。剩余大鼠于P36-P41行Morris水迷宫实验。结果丙泊酚单次或多次暴露均使海马组织TNF-α含量明显高于同期对照水平(P<0.05,P<0.01);P(single)组中,活化caspase-3阳性神经元于P7较对照组同时间点明显增多(P<0.05),而于其它时间点差异无显著性(P>0.05),逃避潜伏期和穿越平台次数与对照组比较均无差异(P>0.05);P(repeated)组中,P13、P21、P35时间点活化caspase-3阳性神经元均明显高于同期对照组水平(P<0.01),逃避潜伏期从d1~d5均明显高于对照组同期水平(P<0.01),且穿越平台次数明显低于对照组(P<0.01);依那西普侧脑室注射后,丙泊酚麻醉后各时点的活化caspase-3阳性神经元、逃避潜伏期及穿越平台次数与对照组比较差异均无显著性(P>0.05)。结论 TNF-α介导了丙泊酚诱发的发育期海马神经元凋亡和远期认知功能障碍,而远期认知功能障碍可能与持续性神经元凋亡有关。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

Trichinellosis in immigrants in Switzerland

We describe a case of trichinellosis diagnosed at the Division of Infectious Diseases, Hospital of Lugano, in January 2009. This case was associated with a cluster of cases and was traced to the consumption of contaminated meat after a wild boar hunt in Bosnia.