牛磺酸拮抗溶血磷脂酸诱导的大鼠血管平滑肌细胞增殖作用

目的　观察牛磺酸 (Tau)对溶血磷脂酸 (LPA)诱导的大鼠血管平滑肌细胞 (VSMC)增殖作用的影响。方法　在培养的大鼠血管平滑肌细胞上 ,测定3 H 胸腺嘧啶核苷(3 H TdR)参入和丝裂素活化蛋白激酶 (MAPK)活性 ;硫代巴比妥酸盐 (TBA)法测脂质过氧化终产物丙二醛 (MDA)含量。结果　LPA (1× 10 -9～ 1× 10 -7mol·L-1)呈浓度依赖性增加VSMC 3 H TdR参入 ,5、10和 2 0mmol·L-1Tau及MAPK抑制剂PD0 980 59预孵育后分别使LPA (1× 10 -7mol·L-1)诱导的3 H TdR参入较单独LPA组减少 15 %、34%、4 8%和 5 4 % (P <0 0 5或P <0 0 1)。LPA增加VSMCMAPK活性 ,5、10和 2 0mmol·L-1Tau预孵育降低LPA(1× 10 -7mol·L-1)刺激的MAPK活性 ,较单纯LPA孵育低14 %、35 %和 4 4 % (P <0 0 5或P <0 0 1)。随着加入LPA的浓度增加 ,细胞MDA含量增加。与 1× 10 -7mol·L-1LPA组比较 ,5、10和 2 0mmol·L-1Tau预处理的细胞较LPA单独孵育组细胞MDA含量分别低 18%、2 7%、4 5 % (P<0 0 5或P <0 0 1) ,LPA诱导细胞MDA生成的作用不受PD0 980 59预孵育的影响 (P >0 0 5 )。结论　LPA呈浓度依赖性促进VSMC增殖和脂质过氧化 ,前者与其细胞内信号传导MAPK途径有关 ,Tau可有效拮抗LPA诱导的VSMC增殖、MAPK激活和脂质过氧化损伤 ,Tau     

甘草次酸对血管平滑肌细胞c-fos mRNA表达及细胞增殖的影响

目的　研究甘草次酸 (GA)对大鼠血管平滑肌细胞(VSMC)原癌基因c fosmRNA表达及细胞增殖的影响 ,探讨GA与血管紧张素Ⅱ (AⅡ )受体结合的可能机制。方法　原代培养大鼠血管平滑肌细胞 ,[3H]TdR参入DNA、MTT分析法、Northernblot。结果　①低浓度GA( 1× 10 -9mol·L-1)及高浓度GA( 1× 10 -5mol·L-1)均能导致c fosmRNA迅速表达 ;1× 10 -5mol·L-1Sar AⅡ能够阻断AⅡ、GA诱导的VSMCc fos表达 ;低浓度和高浓度GA都能促进AⅡ ( 1× 10 -5mol·L-1)诱导的VSMCc fos表达 ;②低浓度GA对细胞的增殖有促进作用 ;随着浓度的升高 ,促增殖作用减弱 ;高浓度GA抑制细胞的增殖。Sar AⅡ单独作用于VSMC ,不影响细胞增殖 ;Sar AⅡ能够阻断AⅡ对细胞增殖的促进作用 ;Sar AⅡ既抑制低浓度GA促细胞增殖作用 ,也消除高浓度GA抑制细胞增殖作用 ;低浓度GA与AⅡ联合作用 ,对细胞增殖的促进作用更加显著 ;高浓度GA与AⅡ联合作用于VSMC ,则可减轻高浓度GA对细胞增殖的抑制作用。结论　GA能激活c fos转录因子 ,从而产生对VSMC的增殖作用 ,因此 ,GA具有AT1受体的激动剂2 0 0 2 0 4 0 3收稿 ,2 0 0 2 0 7 2 3修回作者简介 :张凤云 ,女 ,33岁 ,硕士 ;贺师鹏 ,男 ,67岁 ,教授。研究方向 :酶和受体的拮抗剂样作用     

儿茶素和漆黄素对柠檬酸铁诱导大鼠原代肝细胞损伤的保护作用

目的　分别从中药苞蔷薇根和叶中提取获得 (+ ) 儿茶素 (CAT )和漆黄素 (FIS)两种黄酮 ,研究其对柠檬酸铁致肝细胞铁超载的药理作用。方法　原代大鼠肝细胞加入 50μmol·L- 1柠檬酸铁诱导肝细胞铁超载。致损伤前加入CAT及FIS或损伤后 2 4h ,加入CAT及FIS ,继续培养 2 4h后 ,测定培养液中的超氧化物歧化酶 (SOD )、乳酸脱氢酶(LDH )、谷草转氨酶 (AST)、谷丙转氨酶 (ALT)活性和白蛋白 (ALB)含量。培养 48h后 ,用MTT法测肝细胞的存活率。结果　在致损伤前加入CAT或FIS ,仅见CAT 5× 10 - 5mol·L- 1和FIS 5× 10 - 5 mol·L- 1对大鼠原代肝细胞的恢复作用与模型对照组相比差异有显著性 (P <0 0 5或P <0 0 1)。在致损伤后加入CAT或FIS ,可见CAT 5× 10 - 5 、5× 10 - 6 、5× 10 - 7mol·L- 1组 ,FIS 5× 10 - 5 、5× 10 - 6 、5×10 - 7mol·L- 1组均可不同程度的提高培养液中的SOD活性 ,降低LDH水平 ,抑制ALT、AST的释放 ,提高ALB的含量 ,提高培养液中肝细胞的存活率 ,与模型对照组比 ,差异有显著性 (P <0 0 1,P <0 0 5)。FIS 5× 10 - 5 mol·L- 1组与正常对照组相比 ,对于提高SOD活性、ALB含量 ,差异无显著性 (P >0 0 5)。结论　CAT、FIS对柠檬酸铁所致大鼠原代肝细胞的损伤具有保护作用     

比较鳗鱼降钙素与17β-雌二醇对体外培养成骨细胞增殖与分化的影响

目的比较鳗鱼降钙素(CT)与17β-雌二醇对大鼠体外培养成骨细胞增殖和分化的影响。方法用酶消化法分离培养新生SD大鼠成骨细胞(OB),取第2继代,在培养液中分别加入不同浓度的CT和17β-雌二醇(E2);用MTT法观察OB的增殖能力(A值)及分化功能(ALP)。结果CT组较对照组从1×10-12mol·L-1起A值增加,且呈剂量依赖关系,当浓度≥1×10-9mol·L-1时,差异才有显著意义(P<0.05);E2组较对照组在1×10-9～1×10-7mol·L-1,A值均增加(P<0.001),以1×10-8mol·L-1作用最明显。CT组较对照组从1×10-12mol·L-1起ALP活性均增加,也呈剂量依赖关系,当浓度≥1×10-10mol·L-1差异有显著性意义(P<0.05);E2组较对照组在1×10-9～1×10-7mol·L-1,ALP活性均增加(P<0.05),以1×10-8mol·L-1作用最强。结论鳗鱼降钙素与17-β雌二醇均可促进大鼠体外培养OB增殖和分化。     

甘草酸对血清和组织胺诱发的大鼠气道平滑肌细胞增生的影响

目的　观察甘草酸对血清和组织胺诱发的大鼠气道平滑肌细胞 (ASMC)增生的影响。方法　MTT法、细胞计数法和流式细胞术检测体外培养的ASMC的光密度、细胞数和细胞周期。结果　①在含 10 0g·L-1FCS的培养液中加入 6×10 -5mol·L-1的甘草酸可使A570 的增速增加 ,而加入 384×10 -5mol·L-1和 15 36× 10 -5mol·L-1却使A570 的增速减慢 ;在含 10 -2 mol·L-1组织胺的培养液中加入甘草酸使A5 70的增速减慢 ,浓度越大作用越明显。②在含 10 0g·L-1FCS的培养液中加入 6× 10 -5mol·L-1的甘草酸可促进细胞增生 ,而加入 384× 10 -5mol·L-1和 15 36× 10 -5mol·L-1则抑制细胞增生 ;在含 10 -2 mol·L-1组织胺的培养液中加入甘草酸 ,细胞增生受到抑制 ,浓度越大作用越明显。③在 10 0g·L-1FCS或 10 -2 mol·L-1的组织胺中培养 96h ,随着甘草酸浓度的增加 ,G1期细胞数增多 ,S期和G2 +M期细胞数减少。结论　①对FCS刺激引起的ASMC增生 ,低浓度甘草酸有促进作用 ,高浓度有抑制作用。②对组织胺刺激引起的ASMC增生 ,低浓度和高浓度甘草酸都有抑制作用     

长春瑞滨诱导人肺癌Calu-3细胞凋亡及机制

目的　观察长春瑞滨 (VRB)诱导人肺癌Calu 3细胞凋亡时Bcl 2和半胱天冬酶 3的表达有无变化。方法　以不同浓度的VRB( 2 0 ,40和 60 μmol·L-1)作用于体外培养的人肺癌Calu 3细胞 2 4h后 ,TUNEL法和吖啶橙染色法观察肺癌细胞凋亡形态学特征 ;流式细胞仪检测肺癌细胞凋亡率和肺癌细胞Bcl 2蛋白表达水平 ;以半胱天冬酶 3荧光分析检测试剂盒测定肺癌细胞半胱天冬酶 3活性。结果　VRB( 2 0 ,40和60 μmol·L-1)处理细胞 2 4h ,TUNEL法及吖啶橙染色均观察到典型的凋亡细胞形态学特征。流式细胞仪检测VRB处理的肺癌细胞凋亡率分别为 ( 3 .1±0 .6) % ,( 7.8± 1 .2 ) %和( 1 9.6± 4.3 ) % ,较对照组 (凋亡率为 0 )显著增高且呈剂量依赖性 (P <0 .0 1 ) ;Bcl 2蛋白阳性表达细胞率分别为 ( 3 7.6±6.9) % ,( 2 5 .4±6.2 ) %和( 8.4±2 .5 ) % ,较对照组 ( 4 8.3±7.1 ) %显著降低且呈剂量依赖性 (P <0 .0 5 ) ;肺癌细胞半胱天冬酶 3活性分别为 ( 3 3 2± 1 6) ,( 4 1 7± 1 1 )和 ( 63 1± 2 7)μmol·L-1·h-1,较对照组 ( 1 95±1 2 ) μmol·L-1·h-1显著增高且呈剂量依赖性(P <0 .0 1 )。结论　VRB可以诱导肺癌细胞凋亡 ,抑制Bcl 2表达及增强半胱天冬酶 3活性     

降钙素基因相关肽对糖尿病大鼠血管平滑肌细胞增殖的影响(英文)

目的　研究降钙素基因相关肽 (CGRP)对体外培养的糖尿病大鼠血管平滑肌细胞 (VSMC)增殖的影响。方法　用酶标仪和流式细胞仪检测不同浓度CGRP(1 ,1 0 ,1 0 0nmol·L-1 )对糖尿病大鼠VSMC的增殖的影响。结果　CGRP呈浓度和时间依赖性的抑制糖尿病大鼠VSMC增殖 ,格列本脲 (1 μmol·L-1 )部分阻断CGRP(1 0nmol·L-1 )对培养的糖尿病组VSMC的抗增殖作用 ,使其对VSMC的抑制率由(49 .3± 1 .2 ) %减至 (35 .6± 1 .1 ) % ,吡那地尔 (1μmol·L-1 )可增强其抗增殖作用 ,使其对VSMC的抑制率由 (49.3± 1 .2 ) %增至 (58.3± 3 .9) %。CGRP(1 0nmol·L-1 )可使停留于G0 /G1 细胞增多 ,进入S期和G2 /M期的细胞数目减少 ,吡那地尔 (1 μmol·L-1 )能增强其作用。结论　CGRP对糖尿病大鼠VSMC具有显著的抗增殖作用 ,使处于G0 /G1 期细胞增多 ,S和G2 /M期细胞减少。     

甲氨喋呤对血管平滑肌细胞增殖、迁移和凋亡的影响

目的　观察甲氨喋呤 (MTX)对体外培养的血管平滑肌细胞增殖、迁移和凋亡的影响。方法　体外培养兔胸主动脉血管平滑肌细胞 ,在培养基中加入不同浓度的MTX ,采用细胞计数及检测细胞周期的方法观察MTX对细胞增殖的影响 ,采用细胞刮片的方法观察MTX对细胞迁移的作用 ,采用检测DNA片断化的方法观察细胞凋亡现象。结果　①MTX在 2 5～ 10 0nmol·L-1的浓度范围内呈剂量依赖性地抑制VSMC增殖。② 2 5nmol·L-1及 5 0nmol·L-1MTX显著增加细胞周期中S期细胞的百分含量 ,减低G2 /M期细胞的百分含量 (P <0 0 5 ) ,10 0nmol·L-1MTX显著增加了G0 /G1期细胞的百分含量 (P <0 0 1)。③加入MTX的VSMC对血清刺激的迁移受到抑制。④ 10 0nmol·L-1及 2 0 0nmol·L-1的MTX作用于VSMC后 ,流式细胞仪检测亚二倍体的细胞数高于对照组 (P <0 0 1) ;DNA凝胶电泳可以见到梯形条带 ;TUNEL染色阳性细胞的百分数高于对照组 ,具有统计学差异 (P <0 0 1)。结论　甲氨喋呤能够抑制体外培养的血管平滑肌细胞增殖和迁移 ,并促进细胞凋亡。     

甲硫氨酸脑啡肽对系统性红斑狼疮患者外周血淋巴细胞增殖反应的影响及阿片受体介导机制的研究

目的　研究甲硫氨酸脑啡肽 (Met Enk)对系统性红斑狼疮 (SLE)患者外周血淋巴细胞 (PBL)增殖反应的影响以及纳络酮 (Nal)对其影响的作用。方法　淋巴细胞增殖法。结果　Met Enk(1× 10 -8～ 1× 10 -6mol·L-1)能够促进正常人PBL增殖反应 ,Nal(1× 10 -6mol·L-1)对正常人PBL增殖反应无明显影响 ,但可拮抗Met Enk(1× 10 -6mol·L-1)对正常人PBL增殖反应的促进作用 ;Nal(1× 10 -6mol·L-1)不仅可拮抗Met Enk(1× 10 -6mol·L-1)对SLE患者PBL增殖反应的促进作用 ,而且可直接降低SLE患者PBL增殖反应。结论　内源性Met Enk通过阿片受体参与了SLE患者PBL功能的上调作用     

血管紧张素Ⅱ和氯沙坦对大鼠小动脉平滑肌细胞的基质金属蛋白酶2和9表达的影响

目的　为研究血管紧张素Ⅱ导致血管重构的机制是否与其影响平滑肌细胞的基质金属蛋白酶 2和 9的表达及活性有关。方法　应用RT PCR、Westernblot、明胶酶谱分析等方法观察不同浓度的血管紧张素Ⅱ对体外培养的大鼠肠系膜小动脉平滑肌细胞的金属蛋白酶 2和 9的表达及活性的影响。结果　高浓度的血管紧张素Ⅱ作用VSMC 48h时 ,可以诱导金属蛋白酶 2和 9的表达及活性增加。不同浓度影响程度不同 ,1× 10 - 8mol·L- 1和 1× 10 - 7mol·L- 1的血管紧张素Ⅱ影响最强。 1× 10 - 6 mol·L- 1氯沙坦可以部分阻断血管紧张素Ⅱ对金属蛋白酶 2和 9的诱导作用。结论　血管紧张素Ⅱ可以诱导小动脉VSMC的金属蛋白酶 2和 9的表达及活性增加 ;AT1受体参与介导此过程     

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.

---

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.