伊贝沙坦对肾间质纤维的保护及治疗作用实验研究

目的观察伊贝沙坦对单侧输尿管梗阻大鼠肾间质中PDGF-BB介导的肾间质纤维化的影响。方法制作大鼠肾间质纤维化模型,随机分为假手术组、模型组和伊贝沙坦治疗组,术后第7、14天分别处死各组中6只大鼠,检测大鼠肾功能、尿β2-微球蛋白,用免疫组化方法测定肾组织中α-平滑肌肌动蛋白(α-SMA)和血小板衍生生长因子(PDGF-BB)蛋白表达,动态观察肾脏病理变化。结果与输尿管梗阻核型(UUO)组比较,治疗组肾间质纤维化明显减轻(P<0.05),肾间质中α-SMA、PDGF-BB表达明显减少(P<0.05)。结论伊贝沙坦可通过下调肾间质PDGF-BB表达减轻肾组织间质纤维化。     

血小板衍生生长因子与CD68在单侧输尿管结扎大鼠肾间质中的表达及意义

目的研究血小板衍生生长因子（PDGF-BB）及CD68（巨噬细胞标志物）阳性细胞在单侧输尿管结扎（UUO）大鼠肾间质中的表达。方法制作大鼠肾间质纤维化模型,于术后第7、14天用免疫组化方法检测肾间质中PDGF-BB、α-平滑肌肌动蛋白（α-SMA）的表达及CD68阳性细胞（巨噬细胞）浸润,并进行相关性分析。结果与假手术组比较,模型组肾间质中PDGF-BB、和α-SMA的表达水平明显升高（P〈0.05）,CD68阳性细胞浸润明显增多（P〈0.05）,模型组CD68阳性细胞在第14天时较第7天时减少。α-SMA与PDGF-BBT CD68的表达呈正相关（r1=0.92,P〈0.01,r2=0.98,P〈0.01）。结论PDGF-BB和巨噬细胞参与肾间质纤维化早期改变。     

血小板衍生生长因子及其受体在梗阻性肾病大鼠肾间质中的表达

目的 研究血小板衍生生长因子(PDGF-BB)及受体(PDGFR-β)在单侧输尿管结扎(UUO)大鼠肾间质中的表达、分布和意义.方法 用免疫组化方法检测UUO大鼠肾间质中PDGF-BB、PDGFR-β和α-平滑肌肌动蛋白(α-SMA)的表达、分布状况和相关性分析.结果 与假手术组相比,模型组肾间质中PDGF-BB、PDGFR-β和α-SMA的表达水平明显提高(P＜0.05).α-SMA与PDGF-BB、PDGFR-β的表达呈正相关(r1=0.92,P＜0.01,r2=0.98,P＜0.01).结论 PDGF-BB、PDGFR-β参与肾间质纤维化发生、发展,抑制PDGF及其受体的产生是防治肾间质纤维化的新途径.     

黄芪当归合剂对肾间质纤维化的多靶点抑制作用

目的动态观察复方中药黄芪当归合剂(A&A)对肾间质纤维化大鼠的抗纤维化治疗作用,了解A&A在疾病过程中的起效时间及不同阶段的主要作用环节。方法♂W istar大鼠随机分为对照(Control)、假手术(Sham)、单侧输尿管结扎(UUO)和UUO+A&A治疗组(UAA)。在给药(A&A 12 g.kg-1.d-1)后d 0、3、7、10处死动物,观察肾功能变化,对肾间质细胞浸润、肾小管萎缩和肾间质纤维化情况进行半定量评分。采用免疫组化、ELISA和W estern B lot分别分析肾组织中α-平滑肌肌动蛋白(-αSMA),转化生长因子β1(TGF-β1),纤粘连蛋白(FN)表达。结果与假手术组相比,各时间点的UUO大鼠肾功能受损、病理损害进行性加重,肾小管间质中TGF-β1、α-SMA、FN表达均增高(P<0.01)。A&A治疗后d 3,可观察到其明显减轻UUO大鼠的肾间质炎性细胞浸润、肾小管萎缩及肾间质纤维化(P<0.05),同时可使肾组织中的TGF-β1表达减少(P<0.05),但此时并不影响α-SMA和FN表达。治疗后d 7,A&A的上述治疗作用持续存在,同时进一步使UUO大鼠肾间质的α-SMA表达减少(P<0.05)、FN表达下降(P<0.05)。治疗10 d后,UAA组肾小管萎缩和肾间质纤维化程度、TGF-β1、α-SMA、FN的表达仍较UUO组分别减轻18.95%,28.7%,39.6%,26%。结论在UUO模型中,A&A的肾保护作用从病变早期开始,主要表现为减少炎症细胞反应和TGF-β1表达分泌,随后可减少肾脏固有细胞转化、分化及细胞外基质成分沉积,其作用的高峰时间是在造模后d 3~7,并持续存在至d10,特征是通过减轻肾间质纤维化而改善肾功能。     

肾安胶囊对5/6肾大部切除大鼠肾脏的保护作用

目的探讨肾安胶囊对肾大部切除大鼠肾脏功能的影响。方法将大鼠随机分为假手术组和手术组,术后3天再将手术组大鼠随机分为肾安胶囊组和Nx组。8周后测尿蛋白和血肌酐;取肾组织做病理检查;用免疫组织化学检查肌成纤维细胞标志抗原(α-平滑肌肌动蛋白α-SMA)。结果Nx组大鼠肾脏出现肾小球硬化和肾间质纤维化;和Sham组相比α-SMA表达明显升高;肾安胶囊组肾脏纤维化明显减轻(P<0.05),α-SMA表达比Nx组明显下降。结论肾安胶囊可抑制肌成纤维细胞增殖和浸润,减轻肾脏纤维化从而保护肾功能。     

活性维生素D3抑制UUO模型大鼠肾间质纤维化作用的研究及其对TGF-β1,α-SMA表达的影响

目的:探讨活性维生素D3对单侧输尿管梗阻模型(UUO)大鼠肾小管间质纤维化的抑制作用,研究该病理过程中转化生长因子β1(TGF-β1),α-平滑肌肌动蛋白(α-SMA)的表达变化。方法:36只雄性Wistar大鼠,随机分为三组,活性维生素D3组、假手术组、模型组,每组12只。造模后活性维生素D3组给予罗盖全(骨化三醇)灌胃(3ng/100g),假手术组及UUO模型组给予等量生理盐水灌胃。三组大鼠于给药后第14天,28d分批(n=6)处死,留取造模侧肾脏。用HE染色及Masson染色法观察肾小管间质纤维化程度,用免疫组化法测定TGF-β1,α-SMA在各组大鼠造模侧肾脏的表达情况。结果:免疫组织化学显示:UUO模型组大鼠的肾脏组织中TGF-β1及α-SMA的表达显著高于假手术组(P<0.05),而在活性维生素D3治疗组中,TGF-β1及的α-SMA表达低于UUO模型组(P<0.05),肾脏组织形态学显示:模型组肾间质纤维化程度明显高于假手术组,活性维生素D3组肾间质纤维化程度明显小于模型组(P<0.05)。结论:活性维生素D3能有效抑制UUO大鼠肾间质中TGF-β1及α-SMA的表达,从而减轻UUO大鼠肾间质纤维化程度。     

红景天在肾间质纤维化大鼠模型中的作用及对CTGF、PDGF-B表达的影响

目的:探讨红景天提取物(rhodiola)在肾间质纤维化中的保护作用及其机制。方法:采用单侧输尿管结扎致肾间质纤维化大鼠模型。将大鼠随机分成4组:假手术组、模型组、依那普利组、红景天组。术后第10天处死各组大鼠,经Western蛋白印迹分析方法检测各组结缔组织生长因子(CTGF)、血小板源性生长因子B(PDGF-B)的蛋白表达水平。用免疫组化半定量检测各组肾间质的α-平滑肌肌动蛋白(α-SMA)的表达,Masson染色评定各组肾小管间质损害程度。结果:模型组CTGF、PDGF-B、α-SMA的表达及肾小管间质损伤指数明显高于假手术组(P<0.01),红景天及依那普利治疗组明显低于模型组(P<0.01)。组间比较,CTGF、PDGF-B差异有极显著性(P<0.01),α-SMA、肾小管间质损伤指数无显著性差异(P>0.05)。各项指标作相关分析,CTGF与PDGF-B,α-SMA的表达及肾小管间质损伤指数为正相关关系。结论:红景天可抑制肾间质纤维化,其作用可能与下调CTGF、PDGF-B的表达有关。     

三七总皂苷对腺嘌呤致肾间质纤维化的影响

目的研究三七总皂苷(PNS)对腺嘌呤致大鼠肾间质纤维化的防治作用。方法Wistar雄性大鼠50只,随机分为正常对照组(n=6)、模型组(n=22)和干预组(n=22)。2%腺嘌呤250mg·kg~(-1)·d~(-1)混悬液灌胃,连续21d,制作大鼠肾问质纤维化模型;干预组7d后同时腹腔注射三七总皂苷50mg·kg~(-1)·d~(-1)。磺柳酸比浊法测定量24h尿蛋白,酶联免疫吸附实验检测血清中血小板源性生长因子(PDGF)-BB,HE、Masson染色观察肾脏病理变化,免疫组织化学法观察肾小管间质α-平滑肌肌动蛋白(α-SMA)表达并行半定量分析。结果与对照组比较,在实验d7、14、21、28,模型组和干预组大鼠血清PDGF-BB、24h尿蛋白定量、肾间质纤维化程度及α-SMA均升高(P<0.05或P<0.01),干预组低于模型组(P<0.05或P<0.01)。结论在腺嘌呤致大鼠肾间质纤维化早期阶段给予三七总皂苷可降低其血清PDGF-BB水平、肾间质α-SMA表达,阻滞肾间质纤维化进展。     

CGRP对UUO小鼠肾组织间质纤维化与炎性因子表达的干预作用

目的 探讨CGRP对小鼠单侧输尿管梗阻(UUO)模型肾间质纤维化的影响.方法 选择雄性小鼠36只,建立小鼠UUO模型,前2 d治疗组和模型组分别每天给予CGRP 1 R.0 nmol·kg-1·d-1和同等量0.9%氯化钠溶液静脉注射.观察模型第14天肾小管损害百分比、肾间质纤维化程度、肾间质α-SMA平滑肌肌动蛋白、肾间质巨噬细胞数,单核细胞趋化蛋白-1(MCP-1)以及F4/80 mRNA的表达.结果 CGRP显著减轻肾小管损害和肾间质纤维化(P<0.05);治疗组肾间质巨噬细胞数F4/80和肾间质α-SMA表达显著低于模型组(P<0.05);CGRP显著抑制MCP-1以及F4/80 mRNA的表达(P<0.05).结论 CGRP减少小鼠UUO模型肾间质巨噬细胞浸润、降低MCP-1以及F4/80 mRNA表达、抑制α-SMA的表达,从而减轻肾间质纤维化.     

来氟米特对肾大部切除大鼠肾皮质α-平滑肌肌动蛋白的影响

目的探讨来氟米特对肾大部切除大鼠肾脏功能的影响。方法将大鼠随机分为假手术组和手术组,术后3d再将手术组大鼠随机分为来氟米特组和模型组。8周后测尿蛋白和血肌酐;取肾组织做病理检查;用免疫组织化学检查肌成纤维细胞标志抗原———α-平滑肌肌动蛋白(α-SMA)。结果模型组大鼠肾脏出现肾小球硬化和肾间质纤维化;和假手术组相比α-SMA表达明显升高;来氟米特组肾脏纤维化明显减轻,α-SMA表达比模型组明显下降。结论来氟米特可抑制肌成纤维细胞增生和浸润,减轻肾脏纤维化从而保护肾功能。     

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.     

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.     

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.