OCE对体外培养的视网膜组织的抗氧化作用

目的：观察不同成份的培养液对成年犬视网膜组织进行体外培养的前后相关生化指标的改变，旨在为视网膜移植提供抗氧化能力的保护性方法。方法：将健康成年家犬74眼随机分成六组，Ⅰ（新鲜视网膜组）Ⅱ（常规培养液组）Ⅲ（高浓度OCE组）Ⅳ（低浓度OCE组）Ⅴ（高浓度全脑组织提取液组Ⅵ（低浓度全脑组织提取液组）培养一周。六组视网膜取材后进行SOD、MDA、G－Px,GST,T－AOC生化指标的测定。结果：Ⅱ组与Ⅰ组SOD、G－Px、GST、T－AOC明显下降；DMA值增多非常显著；Ⅲ、Ⅳ组与Ⅱ组比较，T－AOC、SOD、G－Px、GST值显著增高，Ⅳ组明显高于其它各培养组；Ⅲ、Ⅳ组与Ⅱ组比较，MDA值降低明显，Ⅳ组降低最明显。结论：成年犬OCE中含有清除自由基、增强抗氧化能力的特殊物质。并且低浓度OCE对体外培养的视网膜组织抗氧化作用显著。     

米非司酮对体外培养子宫肌瘤细胞作用的形态学观察

目的观察不同浓度米非司酮作用下体外培养的子宫肌瘤细胞的形态学变化,寻找治疗子宫肌瘤的最佳米非司酮浓度。方法光镜及电镜下观察不同浓度米非司酮（10^-8～10^-4mol/L）下体外培养的子宫肌瘤细胞的其形态学变化。结果 10^-8～10^-7mol/L米非司酮对细胞生长无影响,10^-6mol/L以上浓度肌瘤细胞萎缩,10^-4mol/L抑制作用最强。结论米非司酮破坏肌瘤细胞结构,干扰核膜、胞膜、细胞器、及染色体,并与米非司酮浓度成正相关。     

益气明目口服液对光化学损伤大鼠视网膜保护作用的研究

目的:研究益气明目口服液对光化学损伤大鼠视网膜的保护作用。方法:48只SD大鼠随机分为阴性对照组、模型组及益气明目口服液高、低剂量组。除阴性对照组外,各组大鼠接受(1 900±106.9)Lux绿色荧光灯24h持续光照射,制备大鼠视网膜光化学损伤模型。益气明目口服液高、低剂量组分别于光照前7d灌胃给药30、15mL.kg-1.d-1;阴性对照组及模型组予以等量生理盐水灌胃。光照后6h、6d、14d检测视网膜组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量,并进行视网膜透视电子显微镜和光学显微镜的组织学观察。结果:光化学损伤后6d及14d,益气明目口服液高剂量组视网膜中SOD的活性显著高于模型组(P<0.05),益气明目口服液高、低剂量组视网膜中MDA的含量均显著低于模型组(P<0.05);通过视网膜透视电子显微镜和光学显微镜的组织学观察,益气明目口服液高、低剂量组的病理组织学损害较模型组显著减轻。结论:益气明目口服液对视网膜光化学损伤有显著的保护作用。     

盐酸金刚烷胺对兔急性高眼压视网膜神经节细胞损伤的保护作用

随着对青光眼发病机制的深入研究,谷氨酸对视网膜神经细胞(RGC)的毒性作用已逐步被认识。O n ley[1]证实了谷氨酸对新生小鼠RGC毒性作用并称之为“兴奋性毒性”。体外实验也证实了谷氨酸对RGC的毒性作用[2]。在青光眼发病过程中,高眼压和缺血会使眼内谷氨酸的浓度增加,并且和眼     

生姜提取物对大鼠视网膜缺血-再灌注损伤的保护作用研究

目的:研究生姜提取物对大鼠视网膜缺血-再灌注损伤的保护作用。方法:实验分为3组,即正常对照、模型及生姜提取物(400mg·kg-1)组。采用前房灌注生理盐水60min复制视网膜缺血-再灌注模型。复制模型前5天开始,生姜提取物组灌胃生姜提取物,每天分2次给药,模型组大鼠灌胃蒸馏水,复制模型后仍继续原给药方案。于复制模型后12、24、48、72h测定大鼠视网膜中超氧化物歧化酶(SOD)活性和丙二醛(MDA)、一氧化氮(NO)含量的变化;电镜观察视网膜超微结构。结果:与模型组比较,生姜提取物组大鼠视网膜SOD活性显著增强,MDA、NO含量显著减少(P<0.05);视网膜超微结构损伤有所减轻。结论:生姜提取物可调控SOD活性及MDA、NO含量,改善超微结构而保护视网膜。     

一氧化氮合酶抑制剂对体外培养神经细胞损伤作用的研究

一氧化氮(NO)作为一种新型信使,在正常生理条件下发挥有利作用,但是内源性或外源性NO的产生和过量释放则会直接导致神经毒性。研究表明,NO参与了缺血性脑血管疾病发生发展的诸多环节,但其详细机制并不明确。本实验在体外培养大鼠原代神经细胞的基础上,制成NO毒性模型和神经细胞缺血损伤模型,观察选择性诱导型一氧化氮合酶(NOS)抑制剂氨基胍(AG)和非选择性NOS抑制剂L-硝基精氨酸(L-NA)对体外培养神经细胞损伤作用,为治疗脑缺血疾病提供依据。     

黄芪水提液对大鼠体外神经细胞缺氧损伤的保护作用

<正>黄芪是中医常用益气药之一,具有免疫调节、抗菌、利尿、降压、强心等作用。近年来的研究还发现黄芪可延长常压缺氧小鼠存活时间,延长氰化钾(KCN)中毒小鼠(细胞内缺氧)及亚硝酸钠(NaNO2)中毒小鼠(细胞外液缺氧)的存活时间,同时,还可延长断头小鼠张口动作(脑缺氧)的持续时间[1]。体外实验已证实黄芪对体外培养的心肌细胞、血管内皮细胞具有明显的保护作用。     

脐血贴壁细胞体外培养的动态观察

用脐血MNC体外培养方法，观察脐血造血干细胞在细胞因子作用下形成的贴壁细胞，及其在形态学、组织化学和对造血于细胞生长增殖等方面的动态变化。结果显示：培养两周后的脐血贴壁生长细胞形态以上圆形、椭圆形内皮样细胞和巨噬细胞为主，膜分化抗原主要表达髓系抗原。脐血贴壁生长细胞形成后对造血干细胞的生长增殖具有良好支持作用。研究为实行造血基质细胞体外扩增应用于临床重建骨髓造血微环境提供了实验依据。     

茶多酚对脑缺血再灌注损伤的保护作用及在体外抗脂质过氧化和清除自由基作用

目的 :研究茶多酚对脑缺血再灌注损伤的保护作用及抗脂质过氧化和清除自由基的作用。方法 :结扎大鼠双侧颈总动脉及迷走神经后 ,再灌注 4 5min ,建立脑缺血再灌注模型。由自由基诱导的脑线粒体脂质过氧化 ,用硫代巴比妥酸法测定。由Xan XO诱导的O 2 and由Fe2 + H2 O2 诱导的·OH ,用紫外分光光度仪测定。结果 :在脑缺血再灌注损伤时 ,茶多酚能改善SOD、GSH Px、CAT活性 (P <0 0 1) ,同时降低脑水肿和MDA含量 (P <0 0 1和P <0 0 5 )。茶多酚具有明显的清除·OH和O 2 的作用 (IC50 分别为 2 2mmol·L-1和 1 9mmol·L-1) ,茶多酚明显抑制由·OH诱导脑线粒体脂质过氧化 ,并呈浓度依赖性。结论 :结果提示茶多酚能保护脑缺血再灌注损伤 ,与其清除自由基和抗脂质过氧化作用有关     

Three-dimensionally engineered biomimetic tissue models for in vitro drug evaluation: delivery,efficacy and toxicity

Introduction: Three-dimensionally (3D) engineered biomimetic tissue models are sought after due to their high fidelity in mimicking various native tissues of the human body, this quality of which gives them an important role at the forefront of drug discovery and development. A multitude of studies have consistently indicated that gene expression profiles, cellular phenotypes, differentiation capabilities and functionalities are all affected by tissue architecture. Thus, the drug evaluation process will stand to gain immense benefits from the fairly accurate predictions of cellular responses displayed by 3D-engineered tissue models when exposed to the drugs of interest in vitro. Stemming from this fact, many studies have set out to capitalize on developing tissue models that are tailored to specific aspects of drug evaluation including the tests of novel drug delivery systems, drug efficacy and toxicity.

Areas covered: The areas covered include fabrication methods and usage of 3D in vitro tumor models in cancer research, focusing on the evaluation of delivery and efficacy of various anticancer drugs or other therapeutic agents. Also covered are the use of 3D in vitro inflammatory tissue models in anti-inflammation research, centering on osteoarthritis (OA) and rheumatoid arthritis (RA) and the use of 3D in vitro tissue models designed for drug toxicity evaluation specifically with liver-mimetic tissues.

Expert opinion: Currently available 3D tissue models in various fields of research have already displayed their capabilities in predicting cellular responses to various therapeutic agents and delivery methods with better accuracy than their 2D counterparts, albeit being in need of much refinement before they can be successfully applied for reliable drug evaluation. Given further development and improvement, it is highly probable that the 3D-engineered tissue models may perform as living platforms for dynamic drug evaluation in vitro.     

低剂量黄芪提取液联合左氧氟沙星体外抑菌作用及机制研究

本研究采用低剂量黄芪提取液联合左氧氟沙星进行体外抑菌实验,结合黄芪提取液中微量元素含量及细胞膜通透性实验,探讨了低剂量黄芪提取液联合左氧氟沙星在体外对6种临床常见微生物的抑菌作用及其机制。结果表明黄芪提取液联合左氧氟沙星对大肠埃希菌、金黄色葡萄球菌、肺炎克雷伯菌、变形菌和粪肠球菌均表现出协同抑菌作用,而该剂量黄芪提取液本身不具有抑菌力。检测黄芪提取物的Ca、Zn、Mg、Cu和Fe元素浓度分别为590、224.72、300、0.06和0.4μmol/L。细胞膜通透实验结果表明,黄芪提取物处理的菌液K+浓度随时间延长而增加,Na+和Cl-浓度随时间延长而降低。因而推断在该联合抑菌实验中,低剂量黄芪提取物可能通过增强细胞膜通透性以提高左氧氟沙星在细胞内的有效浓度,进而更有效地抑制DNA解旋酶活性达到协同抑菌的目的。     

冬虫夏草对离体人肾小球系膜增殖的影响

目的 :观察冬虫夏草对离体人肾小球系膜增殖的疗效 ,并探讨其机制。方法 :利用体外培养的人肾小球系膜细胞 ,采用检测 3H胸腺嘧啶掺入量的方法 ,研究北冬虫夏草和天然冬虫夏草对低密度脂蛋白 (L DL )引起系膜细胞增殖的影响。结果 :2组冬虫夏草组的 3H掺入率均明显低于 L DL对照组 (P<0 .0 1) ,而 2组冬虫夏草之间同一浓度相比无显著性差异 (P>0 .0 5 )。结论 :冬虫夏草明显抑制 L DL引起的系膜细胞增殖 ,北冬虫夏草与天然冬虫夏草作用相似。其机制可能是由于冬虫夏草间接抑制了系膜细胞增殖过程中 DNA的合成。     

镁与胰岛素对大鼠缺血脑组织的保护作用

目的 探讨镁与胰岛素对缺血脑组织的保护作用。方法 大鼠全脑缺血前后腹腔注射硫酸镁和胰岛素，与对照组比较脑组织含水量、神经细胞计数和形态学观察、超氧化物歧化酶（SOD）和神经原特异性烯醇化酶（NSE）。结果 观察组海马区正常神经元计数和SOD含量显著高于对照组（P分别＜0．01和0．05），脑组织含水量和NSE含量显著低于对照（P分别＜0．01和0．05）。结论 硫酸镁和胰岛素对缺血脑组织有肯定的保护作用。     

Effects of valproic acid,some of its metabolites and analogues on prenatal development of rats in vitro and comparison with effects in vivo

Using a whole-embryo culture system valproic acid (VPA) and some of its metabolites (2-en-VPA, 4-en-VPA, 4,4-dien-VPA) and analogues (ethyl-propyl-acetic acid, propyl-butyl-acetic acid, di-butyl-acetic acid, 2-methyl-2-ethyl-hexanoic acid, 1-methyl-1-cyclohexanoic acid) were tested for their potential to induce abnormal development. With regard to embryonic growth, development and abnormality rate, the tested compounds showed a wide range of teratogenic potency in vitro. In order to verify some of the in vitro results, in vivo experiments were performed. Pregnant rats were treated subcutaneously on day 10 of gestation with 2×330 mg VPA/kg, or 2×400 mg 2-en-VPA/kg, respectively. Evaluation of the embryos was performed on day 11.5 of gestation, corresponding to the in vitro experiments. VPA showed a high potential to induce abnormal development in vivo as well as in vitro, whereas 2-en-VPA was inactive under our experimental conditions. Problems connected with the evaluation of the predictive value of an in vitro test system for the detection of embryotoxic effects, such as validation and significance of pharmacokinetic data, are discussed.Some of the data presented in this paper are part of the doctoral thesis of Frank Zappel to be presented to the Fachbereich Veterinärmedizin, Freie Universität Berlin.     

吡罗昔康普朗尼克磷脂有机凝胶的体外评价和局部组织分布

目的制备普朗尼克磷脂有机凝胶,以吡罗昔康为模型药物,评价其体外性能并考察其局部组织分布特征。方法应用椎-板模型流变仪测定普朗尼克磷脂有机凝胶的流变学参数,Franz扩散池法测试其体外释放和经皮通透性,考察大鼠局部给药的局部组织分布。结果普朗尼克磷脂有机凝胶具有典型的凝胶的流变学特征;48 h体外累积释放率和经皮通透率分别为(81.56±3.09)%和(3.80±1.59)%;皮肤和肌肉的药时曲线具有双峰特征,给药侧的组织药物浓度高于血药浓度和对侧相同组织的浓度,原因在于药物的直接通透和系统再分布。结论普朗尼克磷脂有机凝胶适用于皮肤局部给药的载体。     

榼藤子水溶性提取物的体外抗肿瘤作用

目的观察榼藤子水溶性提取物对3种不同肿瘤细胞株的抑制作用。方法采用噻唑蓝染色法(MTT法),肿瘤细胞株选用人类慢性髓性白血病细胞株(K562)、人类淋巴瘤细胞株(U937)和人早幼粒白血病细胞株(HL60),检测A值,计算抑瘤率。结果榼藤子水溶性提取物对K562、U937、HL60有较强的抑制作用,且呈一定的浓度依赖性,半数生长抑制剂量(IC50)均<20μg·ml-1。结论榼藤子水溶性提取物体外实验具有显著的抗肿瘤活性。     

In vitro accumulation of tetrodotoxin in pufferfish liver tissue slices.

The liver tissue slices of pufferfish accumulate in vitro tetrodotoxin (TTX), when incubated with minimum essential medium containing TTX. In the case of Takifugu rubripes liver slices incubated at a concentration of 25 microg TTT/ml, TTX of 3.9 microg/g was first detected at 2h and increased to 15 microg/g at 48h. The TTX content accumulated was not decreased, even when the slices were further incubated without TTX for additional 48h. Another species of pufferfish T. paradalis also showed similar trend in TTX accumulation, except they accumulated higher concentration of TTX (36.4 microg/g at 48h) than T. rubripes. On the contrary, in the cases of the liver slices from parrot-bass Oplegnathus fasciatus, green ling Hexagrammos otakii and filefish Thamnaconus modestus incubated at a concentration of 25 microgTTX/ml, TTX of 3-4 microg/g was detected even at 0.5h. However, no significant change in TTX contents was recognized during the incubation for 48h. Further incubation of the filefish liver slices without TTX for additional 48h did not decrease the TTX content. It is unlikely that the liver slices of filefish as well as pufferfish rapidly excrete TTX. These results suggest that the difference in the accumulation of TTX between pufferfish and filefish livers is ascribable to the difference not in the TTX excreting ability but in the ability to take up TTX.     

人参皂甙对培养小鼠皮层神经细胞在糖氧剥离损伤中的保护作用

目的 研究人参皂甙（Gs）的三种单体成分GSRb1、GSRb3、GSRg1对离体培养的小鼠皮层神经细胞在糖氧剥离损伤中的保护作用。方法 原代培养的小鼠胎鼠大脑皮层神经细胞分为正常对照组、糖氧剥离模型组、GSRb1、GSRb3、GSRg1干预组。利用糖氧剥离（OGD）建立皮层神经细胞缺血／再灌注损伤模型,分别用60μmol／LGSRb1、GSRb3、GSRg1干预后,再测定各组细胞培养上清液一氧化氮（NO2^-／NO3^-）、乳酸脱氢酶（LDH）、四唑盐（MTT）比色试验测定神经细胞活力,用流式细胞仪检测细胞凋亡率。结果 GSRb1、GSRb3、GSRg1干预组与模型组比较,NO分泌量、LDH漏出量、细胞活力、细胞凋亡率均明显减少（P〈0．01）。结论 GSRb1、GSRb3、GSRg1对培养小鼠皮层神经细胞在糖氧剥离损伤中具有保护作用,其可能机制之一是通过拮抗兴奋性氨基酸毒性,减少NO分泌实现的。