产后抑郁小鼠模型的构建及越鞠甘麦大枣汤对其抑郁样行为的影响

目的建立产后抑郁症(PPD)小鼠模型,评估母鼠产后的异常行为,观察中药越鞠甘麦大枣汤对产后抑郁母鼠的快速抗抑郁作用。方法将32只Balb/c母鼠随机分为2组,空白对照组不给予任何刺激,模型组小鼠给予慢性束缚刺激。3周后予模型组小鼠♀♂合笼交配怀孕,于分娩后3周检测母鼠行为学,包括蔗糖消耗测试(sucrose preference test,SPT)、强迫游泳测试(forced swimming test,FST)及陌生环境摄食实验(novelty suppressed feeding test,NSFT),以评价造模是否成功。造模成功后,模型组单次给予越鞠甘麦大枣汤(越甘组),空白对照组和模型组予以生理盐水,以单次给予模型组西药氯胺酮(氯胺酮组)设为阳性参照组。结果产后3周,与空白对照组比较,在蔗糖消耗测试中,模型组母鼠的糖水消耗百分比明显降低(P<0.01);在强迫游泳测试中,模型组母鼠绝望状态不动时间明显延长(P<0.01);在陌生环境摄食实验中,模型组母鼠摄食潜伏时间明显增加而单位时间摄食量明显减少(P<0.01,P<0.01)。而单次给予中药越甘24 h之后,PPD模型母鼠的糖水消耗百分比明显增加(P<0.01),在强迫游泳测试中的绝望不动时间明显降低(P<0.01),陌生环境摄食测试中的潜伏时间明显缩短,单位时间摄食量明显增加(P<0.01,P<0.01),表现出与西药氯胺酮相似的结果。结论慢性孕前应激可诱导Balb/c母鼠表现出产后抑郁样症状;中药越甘对产后抑郁小鼠模型具有与西药氯胺酮相似的快速抗抑郁作用。     

越鞠甘麦大枣汤对抑郁子代小鼠海马Akt及m-TOR分子表达的影响

目的观察越鞠甘麦大枣汤对子代抑郁症的快速疗效,并分析其对Akt及m TOR分子的影响。方法成功建立产后抑郁子代模型后,随机分组如下:正常子代组(CTL-F1,8只),抑郁子代生理盐水组(Veh,8只)、越鞠甘麦大枣汤组(YG,8只)。Veh组给予生理盐水,YG组给予越鞠甘麦大枣汤(8.3 g·kg~(-1))。单次给药24 h后测量强迫游泳实验(FST)。取脑,Western blot法检测小鼠海马Akt及m-TOR的磷酸化及总体水平。结果 YG组的游泳不动时间比Veh组明显缩短(P<0.01),并且小鼠海马p-Akt和p-m TOR表达明显上调(P<0.05)。结论越鞠甘麦大枣汤可能通过上调Akt和m TOR表达,快速缓解子代抑郁样行为。     

雷公藤内酯醇对应激小鼠抑郁样行为及脑源性神经营养因子的调节作用

目的研究雷公藤内酯醇对抑郁模型小鼠抑郁样行为的改善及脑源性神经营养因子(BDNF)的影响。方法成年雄性ICR小鼠,每天选取8种刺激中的1种进行刺激应激,每日1次,共计14 d,制备慢性不可预知应激抑郁模型。每天应激前10 min,ip给予雷公藤内酯醇20,40,80和160μg·kg-1,第15天起,给予药物处理10 min后,每天检测一个行为学指标。采用自发活动实验测定运动总路程,强迫游泳实验和悬尾实验测定累计不动时间。采用Western蛋白印迹法检测小鼠海马和前额叶中磷酸化c AMP反应元件结合蛋白(p-CREB)和BDNF的蛋白表达。结果雷公藤内酯醇不影响小鼠的自主活动能力。慢性应激组表现出明显的抑郁样行为,给予雷公藤内酯醇80和160μg·kg-1后在强迫游泳实验的不动时间分别由慢性应激组的(160±18)s下降为(102±14)s(P<0.05)和(83±14)s(P<0.01),而丙米嗪(10 mg·kg-1)和氟西汀组(10 mg·kg-1)分别为(77±11)s(P<0.01)和(96±9)s(P<0.01)。给予雷公藤内酯醇40,80和160μg·kg-1后在悬尾实验中的不动时间分别由慢性应激组的(128±8)s下降为(93±9)s(P<0.05),(85±8)s(P<0.01)和(77±11)s(P<0.01),丙米嗪和氟西汀组分别为(64±9)s(P<0.01)和(72±6)s(P<0.01)。雷公藤内酯醇80和160μg·kg-1组海马内p-CREB蛋白表达增加(P<0.05),前额叶内p-CREB蛋白表达亦显著性增加(P<0.05)。雷公藤内酯醇80和160μg·kg-1组海马和前额叶中BDNF的表达显著性增加(P<0.05)。结论雷公藤内酯醇对慢性不可预知应激引起的小鼠抑郁样行为有一定程度的改善作用,且这种改善作用可能与其参与调节p-CREB-BDNF通路有关。     

三氟淫羊藿素对心肌缺血再灌注损伤大鼠Akt/mTOR信号通路及自噬相关蛋白水平的影响

目的研究三氟淫羊藿素(ICTF)对大鼠心肌缺血再灌注损伤(MI/RI)的治疗作用,并探讨其是否通过丝氨酸/苏氨酸蛋白激酶/哺乳动物雷帕霉素靶蛋白(Akt/mTOR)信号通路调节自噬起作用。方法采用结扎左冠状动脉前降支45 min,再灌注60 min的方法制作MI/RI模型,雄性SD大鼠分为假手术组、MI/RI模型组、ICTF 0.5,1.0和2.0 mg·kg~(-1)组。标准肢体Ⅱ导联心电图监测T波和ST段变化,TTC染色测定心肌梗死面积,Western印迹法检测心肌组织中微管相关蛋白2/1轻链3(LC3Ⅱ/LC3Ⅰ)比值、Akt和mTOR蛋白磷酸化水平,免疫荧光检测心肌组织LC3蛋白表达水平。结果监测心电图发现,与假手术组相比,MI/RI模型组大鼠再灌注60 min时的T波(P<0.05)和ST段(P<0.01)明显升高;与模型组相比,ICTF 1.0 mg·kg~(-1)组的T波(P<0.05)和ST段(P<0.01)显著降低。TTC染色结果表明,模型组出现明显的心肌梗死灶(P<0.01),ICTF 1.0 mg·kg~(-1)组心肌梗死面积百分数显著降低(P<0.01)。Western印迹结果显示,与假手术组比较,模型组LC3Ⅱ/LC3Ⅰ比值显著升高(P<0.01),Beclin~(-1)蛋白磷酸化水平升高(P<0.05),Akt(P<0.01)和m TOR(P<0.05)蛋白磷酸化水平降低;与模型组相比,ICTF 1.0 mg·kg~(-1)组LC3Ⅱ/LC3Ⅰ比值明显降低(P<0.01),Beclin~(-1)(P<0.01)蛋白磷酸化水平降低,Akt和m TOR蛋白磷酸化水平升高(P<0.01)。免疫荧光结果表明,与假手术组相比,模型组大鼠心肌组织中LC3蛋白表达增多(P<0.01);与模型组比较,ICTF 1.0 mg·kg~(-1)组表达减少(P<0.01)。结论 ICTF对大鼠MI/RI具有保护作用,其机制可能与调控Akt/m TOR信号通路抑制细胞过度自噬有关。     

淫羊藿苷对产前应激子代大鼠抗抑郁作用研究

目的观察淫羊藿苷对产前应激子代抑郁样行为的干预作用,并探究其作用机制。方法本文采用慢性束缚应激对孕后期母鼠建立抑郁模型,将♂SD子代大鼠随机分为5组:空白对照组(CON)、产前应激组(PS)、生理盐水组(NS)、淫羊藿苷高剂量组(80 mg·kg~(-1))、淫羊藿苷低剂量组(40 mg·kg~(-1))。采用强迫游泳实验和旷场实验对各组大鼠的抑郁样行为进行评价,并通过Western blot测定各组大鼠前额叶皮层区m Glu R1、m Glu R5、EAAT2蛋白的表达。结果与CON组相比,PS组子代大鼠挣扎时间和穿越中央格次数减少(P<0.01,P<0.05),表现出明显的抑郁样行为。PS组较CON组前额叶皮层区mGluR1、mGluR5蛋白表达明显升高(P<0.01,P<0.05),而EAAT2蛋白表达明显降低(P<0.05)。与NS组相比,淫羊藿苷高、低剂量组均能明显增加子代大鼠的挣扎时间(P<0.05,P<0.01)和穿越中央格次数(P<0.05)。淫羊藿苷干预后(80、40 mg·kg~(-1))均能明显改善前额叶皮层区mGluR1、mGluR5及EAAT2蛋白的表达。结论淫羊藿苷对产前应激子代大鼠具有明显的抗抑郁作用,其作用机制可能与脑内Ⅰ族mGluRs的调制作用相关。     

乳岩宁联合依西美坦调控荷瘤裸鼠能量代谢的机制

目的探究乳岩宁联合依西美坦通过PI3k-AKT通路对乳腺癌荷瘤裸鼠能量代谢的调控机制。方法建立乳腺癌(MDA-MB-435)荷瘤裸鼠模型后,分为空白对照组、乳岩宁组、依西美坦组、联合组(依西美坦+乳岩宁组),每天灌胃给药1次,持续21 d。取材,称取各组瘤体质量。采用Western blotting法测定各组PI3k-AKT蛋白的表达。采用实时荧光定量RT-PCR法检测各组m TOR、HIF-1、LDH-A、GLUT1基因表达。结果瘤质量:与对照组比较,乳岩宁组、依西美坦组和联合组的瘤体质量明显降低(P<0.05),其中联合组较中药组和西药组的瘤重有明显降低(P<0.05)。PI3k、AKT的蛋白表达:与对照组相比,中药组、西药组和联合组的PI3k、AKT的蛋白表达均明显下降(P<0.05)。联合组的PI3k和AKT的蛋白表达较中药组和西药组有明显下降(P<0.05)。m TOR、LDH、HIF和GLUT1基因表达:与对照组相比,中药组、西药组和联合组的m TOR、LDH、HIF和GLUT1基因表达均明显降低(P<0.05)。联合组的各项基因表达较中药组和西药组有明显下降(P<0.05)。结论乳岩宁联合依西美坦可能通过阻碍PI3k-AKT通路的激活,抑制下游基因m TOR、HIF-1、LDH-A和GLUT1的表达,以控制肿瘤细胞内糖酵解、蛋白质的合成以及新血管的形成等能量代谢活动,从而达到抑制肿瘤细胞生长的目的。     

迷迭香酸基于AKT/mTOR通路促进抑郁模型大鼠海马神经元再生

目的 探讨迷迭香酸是否基于AKT/mTOR通路促进大鼠海马神经元再生。方法 构建SD大鼠抑郁模型,腹腔注射迷迭香酸或盐酸帕罗西汀28天。应激结束观察各组大鼠行为学指标改变。检测大鼠海马神经生长因子(NGF)、神经元特异性烯醇化酶(NSE)的表达差异及BrdU、NSE的比例。建立皮质酮诱导原代海马神经元细胞损伤模型,通过MK-2206阻断AKT通路,检测给予迷迭香酸后神经元存活率,测定各组AKT、p-AKT、mTOR、p-mTOR蛋白表达。结果 迷迭香酸组大鼠的行为学指标得到显著改善。NGF、NSE的表达显著升高,BrdU与NSE的比例增加。迷迭香酸能够明显改善皮质酮损伤导致的神经元再生,阻断AKT/mTOR后,可拮抗其对神经元的保护作用。结论 迷迭香酸可通过AKT/mTOR通路促进神经元再生,保护海马神经元,发挥抗抑郁作用。     

枸杞籽油对CUS致小鼠应激行为的保护作用

目的探讨枸杞籽油(LBSO)对慢性应激所致小鼠抑郁样行为和认知功能障碍的保护作用及其机制。方法采用甲酯化反应联合GC-MS法测定LBSO的脂肪酸组成和比例。采用孤养结合建立小鼠慢性不可预见性应激模型,实验动物随机分为正常组、模型组、氟西汀组(20 mg·kg~(-1))、LBSO(2. 5、5. 0、10. 0 m L·kg-1)组。旷场、悬尾、强迫游泳和Morris水迷宫实验评价小鼠抑郁样行为和学习记忆能力;ELISA法检测血清皮质酮水平; Western blot法检测海马中BDNF、CREB和p-CREB的蛋白表达。结果本次实验从LBSO样品中分离得到(Z)-9-十八碳烯酸(66. 40%)、反式-9,12-十八碳二烯酸(21. 56%)和十七碳酸(5. 82%)等5个组分。LBSO干预能够提高CUS小鼠旷场实验运动得分,缩短悬尾实验及强迫游泳实验的不动时间,缩短水迷宫实验中的逃逸潜伏期、平均潜伏距离,增加其穿越平台有效次数;降低血清皮质酮水平;增加海马内BDNF蛋白表达及p-CREB/CREB的比值。结论 LBSO可改善慢性应激小鼠的抑郁样行为和认知功能障碍,其机制可能与降低血清皮质酮水平,上调海马内CREB-BDNF信号通路关键蛋白表达有关。     

文拉法辛对慢性应激抑郁模型大鼠脑内ERK1、ERK2磷酸化水平及Egr-1表达的影响

目的:探讨文拉法辛对慢性应激抑郁模型大鼠不同脑区细胞外信号调节激酶(ERK1/ERK2)含量和活性及即刻早期反应因子(Egr-1)的影响。方法:以旷场试验得分评价大鼠行为,将成年SD大鼠随机分为正常对照组、抑郁模型组、抑郁模型+文拉法辛混悬液低、中、高剂量(15、30、60 mg.kg-1.d-1)灌胃组。用慢性轻度不可预见性应激加分养方法建立抑郁大鼠模型,采用蛋白印迹法检测大鼠前额叶、海马、下丘脑、纹状体磷酸化p-ERK1、p-ERK2和非磷酸化ERK1、ERK2蛋白及Egr-1的表达水平。结果:不同剂量文拉法辛组旷场行为得分较给药前有明显升高(P<0.01),而模型组给药前后无改善(P>0.05);除前额叶部位不同剂量文拉法辛组p-ERK1、p-ERK2表达水平较模型组高外,其余各部位各组p-ERK1、p-ERK2、ERK1、ERK2、Egr-1表达水平比较均无显著性差异(P>0.05)。结论:文拉法辛可能通过上调前额叶部位ERK1、ERK2磷酸化水平,明显改善慢性应激抑郁模型大鼠的抑郁样行为。     

远志YZ-50对慢性应激抑郁模型大鼠海马Bax、Bcl-2表达的影响

目的：探讨远志YZ-50对慢性应激抑郁模型大鼠海马神经元凋亡因子Bax、Bcl-2表达的影响。方法：采用健康雄性Ⅱ级Wistar大鼠慢性应激抑郁模型，通过免疫组化染色法检测模型大鼠海马区凋亡调控因子Bax、Bcl-2蛋白的表达，从分子水平探讨在给予远志YZ．50后凋亡调控因子Bax、Bcl-2在慢性应激抑郁大鼠海马组织的变化规律。本实验共设五组，分别是空白对照组、慢性应激模型组、地昔帕明组（20mg·kg^-1）、YZ-50低剂量组（2．8g生药·kg^-1）、YZ-50高剂量（5．6g生药·kg^-1）组。结果：在给予大鼠慢性应激刺激21d，造成应激抑郁模型后，通过检测表明模型组动物Bax蛋白的表达明显增加，Bcl-2蛋白表达很弱；YZ-50低、高剂量组动物Bcl-2蛋白的表达与模型组动物相比明显增加；Bax蛋白的表达与模型组动物相比均显著降低（P〈0．05）。结论：YZ-50能够提高慢性应激抑郁模型大鼠海马区CA3区Bcl-2蛋白的表达、抑制Bax蛋白的表达，调控Bcl-2／Bax比例而抑制神经细胞的凋亡，减少外界刺激对脑部神经元的损害，从而改善其抑郁状态。     

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.