基于蛋白芯片技术探讨丁氏溃结灌肠液对结肠炎大鼠肠道组织差异蛋白表达的影响

目的基于蛋白芯片技术探讨丁氏溃结灌肠液对结肠炎大鼠肠道组织相关靶点的影响,阐释丁氏溃结灌肠液改善肠道炎症反应及肠纤维化的可能机制。方法大鼠随机分为阳性药组、模型组、正常组、丁氏溃结灌肠液组,5只/组。除正常组之外,其他3组均给予3.5%硫酸葡聚糖钠(dextran sulfate sodium,DSS)喂养,结肠炎造模成功后,阳性药组使用美沙拉秦灌肠,丁氏溃结灌肠液组使用丁氏溃结灌肠液灌肠,每天1次,连续4周,停药1 d后,取大鼠结肠黏膜组织,采用GSR-CAA-67抗体蛋白芯片技术检测各组大鼠肠道组织炎症反应及肠纤维化相关蛋白的变化,筛选出各组差异性表达的蛋白。结果通过蛋白芯片检测,发现共有8种有意义的差异蛋白,其中模型组和空白组对照组对比筛选出的8种蛋白包括IFN-γ、促红细胞生成素(erythropoietin,EPO)、TIMP-2、TIM-1、IL-6、TIMP-1、TNF-α、IL-22,差异均有统计学意义(P<0.05)。另一方面,丁氏溃结灌肠液与美沙拉秦均对IL-6、TIMP-1有明显的抑制作用(P<0.05),丁氏溃结灌肠液对TNF-α、IL-22有抑制作用(P<0.05),美沙拉秦组则没有相同作用(P>0.05)。结论丁氏溃结灌肠液具有多靶点的作用,可能通过改善肠道炎症反应及肠纤维化达到治疗溃疡性结肠炎的目的。     

青藤碱对TNBS诱导的慢性小鼠结肠炎的治疗作用

摘 要 目的：评价青藤碱对2,4,6 三硝基苯磺酸（TNBS）灌肠引起的Th1细胞介导的小鼠实验性结肠炎的治疗作用,并探讨其作用机制。方法: 为评价青藤碱对结肠炎的治疗作用,将Balb/c 小鼠随机分为乙醇对照组,TNBS模型组,青藤碱低（50 mg·kg^-1）、中（100 mg·kg^-1）、高（200 mg·kg^-1）剂量组,每组10只。为观察青藤碱对TNBS结肠炎的治疗作用,第1剂TNBS给药后7 d开始灌胃给予青藤碱,持续给药21 d。第28天处死动物,观察结肠黏膜的损伤程度, 测定结肠髓过氧化物酶（MPO）活性,ELISA法分别测定结肠炎症细胞因子 （TNF-α、IL-17和IL-23）蛋白水平。结果: 与TNBS模型组比较,青藤碱中、高剂量组的体质量、大体损伤评分及组织学表现均显著改善（P<0.05）,MPO酶活性显著降低（P<0.05）,结肠黏膜中TNF-α 、IL-17和IL-23的蛋白水平均较TNBS组下降（P<0.05）。结论:青藤碱对TNBS诱导的慢性小鼠结肠炎具有治疗作用,作用机制与青藤碱对Th1炎症细胞因子的抑制有关。     

复方肠泰对葡聚糖硫酸钠诱导的溃疡性结肠炎治疗作用的研究

目的 探讨复方肠泰对葡聚糖硫酸钠(DSS)诱导的小鼠慢性溃疡性结肠炎(UC)的治疗作用。方法 随机将C57BL/6小鼠分为正常对照组、DSS模型组、复方肠泰给药组。采用2% DSS溶液对小鼠进行造模,小鼠连续5 d自由饮用DSS水,接着连续5 d自由饮水,重复3个循环。复方肠泰组每天按照18 g/kg ig给药,其他组ig给予相同体积的蒸馏水。每天记录小鼠体质量、疾病活动指数(DAI)评分结果,采用HE染色观察结直肠组织病理学变化,给予组织病理学评分;ELISA检测TNF-α、IL-10表达水平,免疫组化检测COX-2蛋白表达情况。结果 与DSS模型组比较,复方肠泰组小鼠疾病状态得到明显改善,DAI和组织病理学评分显著降低,炎症因子IL-10表达升高,TNF-α、COX-2蛋白的表达降低。结论 复方肠泰对小鼠慢性UC具有显著地治疗作用,可能通过促进抗炎症因子IL-10的表达,同时抑制促炎症因子TNF-α、COX-2蛋白的表达发挥作用。     

湿生扁蕾口山酮对UC肠纤维化大鼠肠上皮细胞间质转化的影响

目的 探讨湿生扁蕾口山酮对溃疡性结肠炎（UC）肠纤维化大鼠肠上皮细胞间质转化的影响。方法 通过2,4,6-三硝基苯磺酸(TNBS)诱导大鼠溃疡性结肠炎肠纤维化模型,利用湿生扁蕾口 山酮干预溃疡性结肠炎肠纤维化模型大鼠,采用链霉菌抗生物素蛋白-过氧化物酶（S-P）法检测上皮表型标记蛋白E-钙黏蛋白(E-cadherin)和间质表型标记蛋白α-SMA的表达情况,采用实时荧光定量聚合酶链反应(qRT-PCR)法检测E-钙黏蛋白、α-SMA mRNA的表达情况。结果 与模型组相比,湿生扁蕾口山酮显著增加溃疡性结肠炎肠纤维化大鼠肠上皮细胞E-cadherin的表达,抑制α-SMA的表达。结论 湿生扁蕾口山酮可通过抑制结肠上皮细胞间质转化作用,明显改善2,4,6-三硝基苯磺酸诱导的大鼠溃疡性结肠炎肠纤维化。     

康复新液缓解乙酸诱导大鼠急性溃疡性结肠炎及机制研究

目的 研究康复新液对乙酸诱导大鼠急性溃疡性结肠炎（ulcerative colitis,UC）细胞因子的影响,并初步探讨其作用机制。方法 采用乙酸灌肠建立急性UC大鼠模型,除正常对照组及模型对照组外,其余各组分别灌肠给予结肠宁、康复新液2.50,1.25,0.625 g·kg^-1,测定疾病活动指数（disease activity index,DAI）、结肠黏膜损伤指数（colon mucosa damage index,CMDI）及病理组织学评分（histopathological score,HS）,采用酶联免疫吸附法检测大鼠血清中白介素-6（interleukin-6,IL-6）、C反应蛋白（C-reactive protein,CRP）、一氧化氮合酶（nitric oxide synthase,iNOS）以及结肠组织中表皮生长因子（epidermal growth factor,EGF）、环氧合酶-2（cyclooxygenase-2,COX-2）、髓过氧化物酶（myeloperoxidase,MPO）、iNOS的含量。结果 与模型对照组相比,结肠宁及康复新液高剂量能显著降低UC大鼠IL-6、CRP、iNOS、COX-2、MPO表达,升高EGF的表达水平（P<0.01）,改善组织病变情况。结论 康复新液对乙酸诱导的UC有治疗作用,作用机制可能与下调大鼠体内炎症因子的表达量,上调EGF的表达量有关。     

同型半胱氨酸调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的实验研究

目的探讨同型半胱氨酸(homocysteine,Hcy)是否通过调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的机制。方法 SD大鼠分为4组,A组为正常对照组(NS皮下注射+NS灌肠),B组为正常对照+Hcy注射组(Hcy皮下注射+NS灌肠),C组为TNBS模型组(NS皮下注射+TNBS灌肠),D组为TNBS模型+Hcy注射组(Hcy皮下注射+TNBS灌肠)。建立高Hcy血症的实验性结肠炎大鼠模型,实验结束时取大鼠结肠组织病理学检查,并进行结肠匀浆检测MPO活性,采用Western blot方法检测大鼠小肠组织中MEK、ERK、p-ERK、MLCK、p-MLCK的蛋白表达水平,采用RT-q PCR方法检测大鼠小肠组织中MLCK mRNA表达。结果与正常对照组及模型对照组相比,TNBS模型+Hcy皮下注射组大鼠DAI及HI评分增高,结肠匀浆MPO活性增高,小肠黏膜组织MEK、ERK、p-ERK、MLCK、p-MLCK蛋白表达水平增加,MLCK mRNA相对表达量增加。结论 Hcy增加实验性结肠炎大鼠肠黏膜通透性,可能与调控MEK-ERKMLCK信号通路有关。     

大蒜素对大鼠溃疡性结肠炎bcl 2、Bax蛋白表达的影响

目的:观察大鼠溃疡性结肠炎淋巴细胞凋亡及其调控蛋白bcl 2和Bax的表达及大蒜素对其的影响,探讨大蒜素对溃疡性结肠炎肠黏膜的保护作用及其机制.方法:SD大鼠48只随机分为对照组、三硝基苯磺酸组(TNBS组)、三硝基苯磺酸+0.9%氯化钠注射液组(TNBS＋NS组)、三硝基苯磺酸+大蒜素组(TNBS＋Alc组)共4组,每组12只.用TNBS 150 mg&#8226;kg 1灌肠建立大鼠溃疡性结肠炎模型,第2天开始,TNBS＋NS组和TNBS＋Alc组分别以0.9%氯化钠注射液4 mL&#8226;d 1、大蒜素30 mg&#8226;kg 1&#8226;d 1对其进行灌胃,qd,至3周后处死动物.利用DNA缺口末端标记技术(TUNEL法)和bcl 2、Bax蛋白免疫组化染色,分别检测溃疡性结肠炎大鼠肠组织中的淋巴细胞凋亡和淋巴细胞bcl 2和Bax的表达,并观察肠管大体形态和组织学改变.结果:与TNBS组及TNBS＋NS组相比,TNBS＋Alc组中淋巴细胞凋亡增加(P＜0.01),bcl 2表达阳性淋巴细胞减少(P＜0.01).损伤指数明显下降(P＜0.01).结论:大蒜素可以通过促进淋巴细胞凋亡而对TNBS诱导的溃疡性结肠炎大鼠肠黏膜有保护作用.     

雷公藤甲素对实验性大鼠结肠炎组织中细胞因子IFN-γ、IL-12表达的影响

目的探讨雷公藤甲素对TNBS诱导大鼠结肠炎的疗效及对炎性细胞因子IL-12和IFN-γ表达的影响。方法将18只成年SD大鼠随机分为对照组、模型组及雷公藤甲素组,观察大鼠的一般情况,比较结肠组织炎症程度。HE染色观察大鼠结肠组织形态学变化。用反转录聚合酶链反应(RT-PCR)和Western blot(WB)法检测大鼠结肠组织中IL-12、IFN-γ的表达水平。结果对照组大鼠结肠组织中IL-12和IFN-γ的表达水平最低,与对照组相比,模型组大鼠结肠组织中IL-12、IFN-γ表达显著增高(P<0.01),与模型组相比,雷公藤甲素组TL-12、IFN-γ表达明显降低(P<0.05)。雷公藤甲素治疗对大鼠结肠组织及大体变化都有一定程度的改善。结肠组织IFN-γ和IL-12的阳性细胞表达与结肠炎病情轻重呈正相关。结论雷公藤甲素能有效治疗TNBS诱导的大鼠结肠炎,其作用机制可能与雷公藤甲素下调大鼠结肠黏膜炎性细胞因子IL-12和IFN-γ的表达,抑制炎症的发生发展有关。     

环孢素A对DSS结肠炎小鼠肠黏膜通透性影响及其机制的研究

目的探讨环孢素A(CsA)对葡聚糖硫酸钠(DSS)结肠炎小鼠肠黏膜通透性的影响及作用机制。方法 C57BL/6J小鼠(6～8周龄、体质量20 g±2 g、♂),实验分为正常对照组(饮用无菌蒸馏水+腹腔注射0.9%NS)、DSS模型组(DSS溶液+腹腔注射0.9%NS)、环孢素A组(DSS溶液+腹腔注射CsA)。小鼠自由饮用5%DSS溶液1周制备结肠炎模型,环孢素A(0.025 mg.g-1)腹腔注射给药,每天1次,共7 d。每日行DAI评分,实验结束后取结肠进行HE染色评分,结肠匀浆检测MPO活性、TNF-α、IFN-γ、IL-13和IL-17水平,另取小鼠小肠黏膜进行透射电镜检查和肌球蛋白轻链激酶(MLCK)活性测定,采用Evans blue和异硫氰酸荧光素-葡聚糖(FITC-D)方法检测小肠黏膜通透性。结果与正常对照组比较,模型对照组小鼠1周后出现明显体重减轻、便血和腹泻,DAI评分和HI评分增高,同时结肠黏膜MPO活性明显增高。电镜检查小鼠回肠黏膜上皮绒毛萎缩、排列不规则,细胞间连接复合体缩短、变宽,细胞间隙扩大,小肠黏膜通透性增高。小鼠结肠匀浆中TNF-α、IFN-γ、IL-13和IL-17水平均有不同程度增高,小肠黏膜中MLCK活性明显增高。与DSS模型组比较,环孢素A组小鼠DAI评分和HI评分明显降低,结肠黏膜MPO活性减低,回肠黏膜上皮细胞绒毛排列整齐,小肠黏膜通透性降低,小肠黏膜MLCK活性和结肠匀浆中TNF-α、IFN-γ、IL-13和IL-17水平均有不同程度降低。结论环孢素A具有明显的抗DSS小鼠结肠炎作用,机制可能与通过调控MLCK表达,改善肠黏膜通透性有关。     

熊胆粉改善大鼠实验性结肠炎及其作用靶点预测

目的:探究熊胆粉对3-硝基苯磺酸(3-nitrobenzenesulfonic acid, TNBS)诱导大鼠实验性结肠炎的治疗作用，结合网络药理学和分子对接的方法，预测熊胆粉治疗炎症性肠病的药效物质、作用靶点和潜在通路。方法:将大鼠随机分正常组，模型组，阳性药组，熊胆粉高，中，低组。利用TNBS灌肠诱导大鼠实验性结肠炎，评估疾病活动指数(disease activity index, DAI)分数，测量结肠长度和病理改变，ELISA法测定结肠组织TNF-α、IL-1β、IL-6、IL-10炎症因子含量，然后通过UPLC-Q Exactive Orbitrap-MS/MS分析熊胆粉成分，利用相关数据库，预测其可能的信号通路，并对二者的亲和力进行预测。结果:与模型组相比，熊胆粉各剂量组DAI分数显著降低(P<0.01),结肠长度不同程度增加，低剂量组更为显著(P<0.05);结肠组织的炎性细胞浸润减少，黏膜上皮基本完整，腺体排列较规则。检测相关炎症因子，与模型组相比，熊胆粉低剂量组TNF-α含量显著下降(P<0.05),IL-10含量显著上升(P<0.01),熊胆...     

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.