还原型谷胱甘肽片改善酒精性肝损伤的作用及机制研究*

目的：探讨还原性谷胱甘肽片对酒精性肝损伤小鼠模型的保护作用及机制。方法：60只雄性ICR小鼠均分6组：空白组、模型组、水飞蓟宾组、还原型谷胱甘肽片（200、400、800 mg·kg-1）3个剂量组。采用50%的乙醇溶液连续灌胃小鼠6周的方法,构建小鼠酒精性肝损伤动物模型,采用还原性谷胱甘肽片灌胃治疗模型小鼠,检测小鼠体质量、肝脏指数、血清生化指标（ALT、AST、ALP、TC、TG、LDL-c、HDL-c）、肝脏炎症（TNF-α、HMGB-1、IL-6）基因表达水平及肝脏脂滴空泡数,研究还原性谷胱甘肽片的保肝作用。结果：与空白组相比,模型组肝脏指数、血清生化指标（ALT、AST、ALP、TC、TG、LDL-c）、肝脏炎症（TNF-α、HMGB-1、IL-6）基因表达和因子水平均显著升高（P<0.01、P<0.05）,血清中HDL-c水平显著下降（P<0.01、P<0.05）,脂滴空泡数显著增加（P<0.01）;还原型谷胱甘肽片（200、400、800 mg·kg-1）治疗后,模型动物肝脏指数血清（ALT、AST、ALP、TC、TG、LDL-c）、肝脏（TNF-α、HMGB-1、IL-6）基因表达和因子水平均显著降低（P<0.01、P<0.05）,血清中HDL-c水平显著升高（P<0.01、P<0.05）,肝脏细胞病理状态改善,脂滴空泡数显著减少（P<0.01、P <0.05）。结论：还原性谷胱甘肽片能够对酒精性肝损伤小鼠模型具有保护作用,其机制可能与减轻肝脏炎症和减少脂滴沉积有关。     

清肝抑脂汤对慢性肝损伤保护作用的研究

目的：探索清肝抑脂汤（QYT）抗慢性肝损伤的保护作用的影响。方法将小鼠随机分为空白对照组,模型组,阳性药物组,清肝抑脂汤高剂量组,清肝抑脂汤低剂量组。首先建立慢性肝损伤模型,除空白对照组外,其余各组小鼠分别腹腔注射10%CCl4溶液,0.1ml/10g,每周2次,共6周。在建模的同时,各组小鼠分别给予相应的药物治疗。末次给药后次日摘眼球取血,分离血清,检测小鼠血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）酶的活性;同时取肝脏,常规病理切片,观察肝组织病理学变化。结果肝组织病理切片显示清肝抑脂汤能改善肝损伤程度;清肝抑脂汤能显著降低小鼠血清ALT（P<0.05,P<0.01）和AST（P<0.05,P<0.01）酶活性。结论清肝抑脂汤能显著降低受试小鼠血清ALT,AST及ALP酶活性,清肝抑脂汤对CCl4所致小鼠慢性肝损伤具有明显的保护作用。     

五味子护肝片对小鼠急性酒精性肝损伤的保护作用

目的观察五味子护肝片对急性酒精性肝损伤小鼠肝脏的保护作用。方法通过急性酒精性肝损伤小鼠模型,观察五味子护肝片连续给小鼠灌胃30d后小鼠血清谷草转氨酶（AST）、谷丙转氨酶（ALT）、甘油三酯（TG）、极低密度脂蛋白（VLDL）和肝组织中超氧化物歧化酶（SOD）、丙二醛（MDA）水平的变化。结果五味予护肝片中、高剂量组均可降低血清中ALT、AST、TG、VLDL活性或含量,升高肝脏组织s0D活性、降低MDA含量,与肝损伤模型组比较差异有显著性（P〈0．05或0．01）;肝组织病理学改变较肝损伤模型组显著减轻（P〈0．05）。结论五味子护肝片对急性酒精性肝损伤小鼠肝脏有保护作用。     

复方苁蓉益智胶囊联合奥拉西坦治疗轻中度血管性痴呆的临床研究

目的 探讨还原型谷胱甘肽片对酒精性脂肪肝（AFLD）斑马鱼模型的保肝功效。方法 选取黑色素等位基因突变型（albino）品系斑马鱼,分别水溶给予还原型谷胱甘肽片500、1 000、2 000、4 000、8 000 μmol/L,同时设置对照组和模型组,除对照组外,利用2%无水乙醇诱导斑马鱼32 h建立AFLD模型,观察记录斑马鱼的死亡情况及表型,确定还原型谷胱甘肽片的最大耐受浓度（MTC）。黑色素等位基因突变型（albino）品系斑马鱼分别给予还原型谷胱甘肽片500、1 000、2 000 μmol/L和阳性对照药RU-21 100 μg/mL,同时设置对照组和模型组,除对照组外建立AFLD模型,采用油红O染色观察斑马鱼肝脏脂肪染色强度（S）,计算还原型谷胱甘肽片的保肝功效;H&E染色观察斑马鱼肝脏病理学改变;实时荧光定量PCR（qRT-PCR）检测白细胞介素（IL）-6、高迁移率族蛋白1b（HMGB-1b）和肿瘤坏死因子（TNF）-α mRNA表达。结果 还原型谷胱甘肽片对AFLD模型斑马鱼的MTC为2 000 μmol/L。与对照组比较,模型组S显著增加（P<0.001）;与模型组比较,还原型谷胱甘肽片1 000、2 000 μmol/L组和RU-21组S显著降低（P<0.01、0.001）;还原型谷胱甘肽片500、1 000、2 000 μmol/L和RU-21 100 μg/mL组保肝功效分别为14%、32%、71%和48%。还原型谷胱甘肽片2 000 μmol/L组肝细胞结构正常,脂肪空泡较模型组减少。与对照组比较,模型组IL-6、HMGB-1b和TNF-α mRNA表达显著增加（P<0.05、0.01）;与模型组比较,还原型谷胱甘肽片500、1 000、2 000 μmol/L组IL-6、HMGB-1b和TNF-α mRNA表达显著降低（P<0.01、0.001）。结论 还原型谷胱甘肽片对AFLD具有明显的保肝作用,其机制与下调IL-6、HMGB-1b和TNF-α表达有关。     

木通皂苷D对CCl4致小鼠急性肝损伤的保护作用

目的研究木通皂苷D对CCl4致小鼠急性肝损伤的保护作用。方法采用CCl4急性肝损伤模型,小鼠ig给予不同剂量的木通皂苷D(1、0.5、0.25 g·kg-1),并以水飞蓟素(0.2 g·kg-1)为阳性对照药,采用比色法检测小鼠血清的天冬氨酸转氨酶(AST)、丙氨酸氨基转移酶(ALT)及肝脏中还原性谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)的含量。结果各剂量木通皂苷D能显著降低CCl4所致小鼠血清中AST、ALT的水平(P<0.01),同时升高肝脏组织中GSH、SOD的水平(P<0.05),降低肝脏组织中MDA的含量(P<0.05)。结论木通皂苷D对CCl4所致小鼠急性肝损伤具有显著的保护作用,其机制可能与抗氧化作用有关。     

人参皂苷对酒精性肝损伤的保护作用研究

目的 研究人参皂苷提取物(GE)对酒精性肝损伤的保护作用与作用机制.方法 将ICR雄性小鼠分为正常对照组、阳性对照组(联苯双酯)、模型组及人参皂苷提取物高、中、低剂量组,建立慢性酒精性肝损伤模型,用人参皂苷提取物进行干预,ig给药30 d后,称取肝质量,计算肝脏系数,测定小鼠血清丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)活力、三酰甘油(TG)、肝脏内丙二醛(MDA)和还原型谷胱甘肽(GSH)的含量,HE染色观察小鼠肝脏病理变化,综合评价人参皂苷提取物对小鼠酒精性肝损伤的保护作用.结果 人参皂苷提取物各剂量组能明显降低酒精性肝损伤小鼠的肝脏系数(P <0.05)、血清TG的含量(P <0.01),可降低其血清中ALT、AST活力(P <0.05或P <0.01),但与正常组ALT、AST活力仍有差异(P <0.05);可在一定程度上降低肝组织中MDA含量(P <0.01),增高肝组织中GSH含量(P <0.01),减少肝组织病理损伤.结论 人参皂苷提取物对酒精性肝损伤有一定保护作用,其机制可能与抑制肝内脂肪堆积,抗氧化作用有关.     

枸杞多糖对小鼠酒精性肝损伤的保护作用及机制研究

目的 研究枸杞多糖对小鼠酒精性肝损伤的保护作用及机制。方法 将小鼠随机分为对照组、模型组和枸杞多糖低、中、高剂量（75、150、300 mg/kg）组,第1~9天于每日13：00时分别ig给药,模型组和对照组给予等量双蒸水。第10~16天给药4 h后,枸杞多糖和模型组小鼠均ig 50%酒精20 mL/kg进行造模,对照组小鼠给予等量双蒸水。观察小鼠一般状态,末次ig酒精16 h后处死小鼠,检测肝脏指数;全自动生化分析仪检测血清中丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、三酰甘油（TG）、总胆固醇（TC）水平;试剂盒法测定肝组织丙二醛（MDA）、还原型谷胱甘肽（GSH）、谷胱甘肽过氧化物酶（GSH-Px）、总超氧化物歧化酶（SOD）及炎性因子肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）的含量;HE染色观察肝组织病理变化。结果 与模型组比较,枸杞多糖各剂量组小鼠醒酒时间短,毛色有所改善,较活跃;各剂量组肝脏指数呈下降趋势,但不具有统计学意义;各剂量组血清ALT、AST、TC、TG均呈下降趋势,其中高、中剂量组ALT显著降低（P<0.05）,3个剂量组TG浓度均差异显著（P<0.01）;各剂量组小鼠肝脏MDA含量显著降低（P<0.05、0.01）,GSH、SOD水平显著升高（P<0.05、0.01）,GSH-Px水平升高但未表现出显著性差异;高、中剂量组小鼠肝脏TNF-α和IL-1β水平显著降低（P<0.05、0.01）。HE染色显示,与模型组比较,枸杞多糖各组肝组织破坏程度较轻。结论 枸杞多糖对于乙醇诱导的酒精性肝损伤具有一定的保护作用,作用机制可能与通过清除体内多余自由基、增强体内抗氧化能力以及减轻炎症反应相关。     

藏药蕨麻对实验性酒精肝损伤小鼠的保护作用研究

目的：探讨藏药蕨麻对实验性酒精肝损伤小鼠的保护作用。方法：50只小鼠随机分为5组：空白组,模型组,小、中、大剂量蕨麻组,50%酒精灌胃复制实验性酒精肝损伤模型,蕨麻每日2次（50、100、200g/kg）,酒精每日1次（10mL/kg）,连续4周。小鼠最后1次给药4h后,称重,眶静脉丛取血,离心取血清,全自动生化仪检测肝功指标[AST、ALT、碱性磷酸酶（ALP）];摘取肝脏称重,计算肝脏指数;剪取适量肝脏制成匀浆,检测肝脏组织中TG、TC、丙二醛（MDA）、脂质过氧化物（LPO）含量以及超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、过氧化氢酶（CAT）活性;剩余肝脏10%甲醛固定后进行病理学检测。结果：小鼠酒精肝损伤后肝脏指数、AST、ALT、ALP明显升高（P〈0.05）;肝脏组织中TG、TC、MDA、LPO含量明显高于空白组（P〈0.05）;SOD、GSH-Px、CAT活性明显降低（P〈0.05）;病理学检测发现肝脏组织出现炎性浸润和脂肪空泡。蕨麻可明显改善这些现象,表现为降低肝脏指数和肝功指标（P〈0.05）,减少肝脏组织中TG、TC、MDA、LPO含量（P〈0.05）,增强SOD、GSH-Px和CAT活性（P〈0.05）,减少肝脏组织炎性浸润和脂肪空泡的产生。其中大剂量蕨麻的作用更加明显。结论：蕨麻可能通过减少自由基的产生,增强对自由基及其代谢产物的清除能力,从而抑制脂质过氧化,起到对酒精性肝损伤的保护作用。     

还原型谷胱甘肽与多烯磷脂酰胆碱对环磷酰胺诱导小鼠肝损伤的修复作用

目的：分析比较还原型谷胱甘肽与多烯磷脂酰胆碱对环磷酰胺诱导小鼠肝损伤的防护作用。方法采用简单随机抽样法将40只昆明小鼠分为肝损伤模型组、还原型谷胱甘肽组、多烯磷脂酰胆碱组和对照组,每组10只。前3组小鼠实验第1-4天均腹腔注射环磷酰胺（100 mg· kg-1·d-1）诱导肝损伤,第5-14天分别腹腔注射0.9%氯化钠注射液0.2 ml、还原型谷胱甘肽180 mg · kg-1· d-1、多烯磷脂酰胆碱90 mg·kg-1·d-1;对照组同期腹腔注射等体积0.9%氯化钠注射液。实验第1天给药前和第15天测定小鼠体重;实验第15天,小鼠处死前眼眶取血测定血清总胆红素和谷胱甘肽水平,处死小鼠后取肝脏称重并计算肝脏系数,取肝组织测定丙氨酸转氨酶（ ALT）、天冬氨酸转氨酶（AST）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性和丙二醛（MDA）水平,并进行肝组织形态学观察。结果实验第15天,应用环磷酰胺的3组小鼠体重均明显低于对照组（P 〈0.01或 P 〈0.05）,但还原型谷胱甘肽组体重高于肝损伤模型组（P 〈0.05）;肝损伤模型组小鼠肝脏系数（5.74%±0.11%）高于对照组（4.68%±0.37%）和还原型谷胱甘肽组（4.81%±0.19%）（均 P 〈0.01）,多烯磷脂酰胆碱组小鼠肝脏系数（5.25%±0.35%）]也高于对照组（P 〈0.05）。肝损伤模型组、还原型谷胱甘肽组、多烯磷脂酰胆碱组血清总胆红素水平[（129.8±1.9）、（110.9±1.3）、（125.7±2.6）μmol/ L]均高于对照组（100.8±3.0）μmol/ L（均 P 〈0.01）,但还原型谷胱甘肽组低于肝损伤模型组（P 〈0.01）。肝损伤模型组和多烯磷脂酰胆碱组血清谷胱甘肽水平[（50.5±1.9）、（55.9±2.4）g/ L]均低于对照组和还原型谷胱甘肽组[（73.8±4.3）、（71.3±3.7）g/ L]（均 P 〈0.01）。肝损伤模型组、还原型谷胱甘肽组、多烯磷脂酰胆碱组肝组织 AST、ALT、SOD 和 CAT ?     

川穹嗪注射液对小鼠急性酒精性肝损伤的保护作用观察

目的：观察川穹嗪注射液对小鼠酒精性肝损伤的保护作用。方法利用50％的酒精建立急性肝损伤小鼠模型,分别以川穹嗪注射液按50、100和200 mg／kg小鼠腹腔注射给药,检测其对血清中天门冬氨酸氨基转移酶（ AST）、丙氨酸氨基转移酶（ ALT）;肝组织丙二醛（ MDA）、还原型谷胱甘肽（ GSH）含量的影响,并用苏木素－伊红（ HE）染色法观察肝组织病理学变化。结果急性酒精性肝损伤小鼠动物模型构建成功,川穹嗪注射液能够可明显降低酒精致肝损伤小鼠血清ALT和AST水平（P＜0．05）,降低小鼠肝脏组织丙二醛（MDA）含量的升高（P＜0．05）,同时使还原型谷胱甘肽（GSH）含量升高（P＜0．05）。减轻肝脏水肿、坏死等病理组织损伤。结论川穹嗪注射液对小鼠酒精性肝损伤具有保护作用,其作用机制可能与抑制氧化活性有关。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.