巴戟天正丁醇提取物对乳鼠心肌细胞缺氧/复氧损伤后炎性细胞因子的影响

目的：观察巴戟天正丁醇提取物对乳鼠心肌细胞缺氧／复氧损伤后炎性细胞因子的影响。方法：采用培养的乳鼠心肌细胞建立缺氧／复氧损伤模型，实验分正常细胞培养组、缺氧／复氧损伤模型组、巴戟天正丁醇提物小、中、大剂量组。平衡法测定TNF-α、IL-1β、IL-6的含量变化。结果：巴戟天正丁醇提物可明显改善缺氧、复氧损伤；降低TNF-α、IL-1β、IL-6的含量。结论：巴戟天正丁醇提取物对缺氧／复氧损伤的心肌细胞有防护作用；其防护作用可能与降低TNF-α、IL-1β、IL-6的含量有关。     

麝香保心丸抗心肌细胞缺氧-复氧损伤活性成分筛选

目的：建立原代心肌细胞缺氧-复氧模型,并对麝香保心丸及其20种血中活性成分进行抗缺氧-复氧损伤的活性筛选。方法取新生（1～3 d）SD乳鼠的心脏,建立原代心肌细胞的缺氧-复氧损伤模型,并采用四甲基偶氮唑盐（MTT）比色法进行麝香保心丸及其入血成分的活性筛选。结果麝香保心丸在50μg/ml浓度下具有较好的保护原代心肌细胞缺氧-复氧损伤的作用;人参皂苷Rb1、人参皂苷Rb2、蟾毒灵和麝香酮具有较好的保护原代心肌细胞缺氧-复氧损伤的作用。结论麝香保心丸具有抗心肌细胞缺氧-复氧损伤的作用,人参皂苷Rb1、人参皂苷Rb2、蟾毒灵和麝香酮是其主要的有效成分。该研究为麝香保心丸深入的药效和机制研究奠定了基础。     

茜草双酯对缺氧/复氧心肌细胞的保护作用研究

目的探讨茜草双酯对缺氧复氧的乳鼠心肌细胞是否具有保护作用。方法原代培养乳鼠心肌细胞,建立缺氧复氧模型,分别测各组细胞乳酸脱氢酶(LDH)活性、丙二醛(MDA)含量及细胞的凋亡与坏死。结果与缺氧/复氧模型组比较发现,缺氧/复氧模型预先加茜草组较缺氧/复氧模型组LDH活性和MDA含量降低有统计学意义(P<0.05),细胞凋亡和坏死有所减少。结论茜草双酯对体外培养的缺氧/复氧心肌细胞具有保护和抗凋亡作用。     

蒺藜总皂苷对缺氧复氧心肌细胞TNF-α,IL-1β及ICAM-1表达的影响

目的:观察蒺藜总皂苷(gross saponins from Tribulus terrestris L,GSTT)对缺氧复氧心肌细胞炎症分子肿瘤坏死因子-α(TNF-α),白介素1β(IL-1β)及细胞间黏附分子-1(ICAM-1)表达的影响。方法:利用原代培养的SD乳鼠心肌细胞建立心肌缺氧复氧模型,分为正常组、模型组和GSTT高、低浓度(50,10mg·L~(-1))组。用放免法检测TNF-α和IL-1β,细胞ELISA检测ICAM-1表达。结果:与模型组比较,GSTT高、低浓度(50,10mg·L~(-1))组TNF-α和IL-1β含量显著降低(P＜0．01或P＜0．05);GSTT 50 mg·L~(-1)组ICAM-1蛋白表达量显著降低(P＜0．05)。结论:GSTT对缺氧复氧损伤心肌细胞有保护作用,其机制与抑制炎性因子TNF-α和IL-1β释放,降低ICAM-1蛋白表达有关。     

红景天苷衍生物N03对乳鼠心肌细胞缺氧-复氧损伤的保护作用

目的：研究红景天苷衍生物1-C-β-D-吡喃葡萄糖基-N-（2′-氯苯甲基）甲胺（1-C-β-D-glucopyranosyl-N-(2’-chlorophenyl) methyl methanamine,N03）对乳鼠心肌细胞缺氧-复氧损伤的保护作用。方法：以1~2 d龄Wistar大鼠乳鼠心室肌进行心肌细胞培养并建立缺氧-复氧损伤模型,检测不同浓度（2,10,50 mg&#8226;L-1）N03预处理后细胞的形态变化,比色法检测乳酸脱氢酶(LDH)、肌酸激酶（CK）、天冬氨酸转移酶（AST）的泄漏量及超氧化物歧化酶(SOD)活性,luminometer法检测细胞中三磷酸腺甘(ATP)水平,荧光探针负载法检测其是否能够拮抗缺氧-复氧损伤所引起的细胞内活性氧(ROS)升高和钙超载。结果：红景天苷衍生物N03能减轻心肌细胞缺氧-复氧损伤后LDH,CK和AST的漏出量,同时能够升高SOD活性,降低缺氧-复氧损伤所致的ATP水平下降程度和拮抗细胞内ROS升高和钙超载。结论：红景天苷衍生物N03对缺氧-复氧损伤的心肌细胞具有明显的保护作用,可能对心肌缺血的一些疾病具有治疗作用     

附子多糖对缺氧/复氧乳鼠心肌细胞的保护机制

目的以细胞凋亡的线粒体信号通路为着眼点,对附子多糖对缺氧/复氧后心肌细胞的保护机制进行探讨。方法建立乳鼠心肌细胞缺氧/复氧模型,将乳鼠心肌细胞分为正常对照组、缺氧/复氧组、缺氧后适应组和附子多糖组。缺氧/复氧组给予心肌细胞缺氧3 h后复氧6 h;缺氧后适应组在细胞缺氧3 h后,复氧前即给予3个循环的5 min复氧/5 min缺氧,随后复氧6 h;附子多糖组在缺氧3 h后,将心肌细胞换入含附子多糖浓度为10 g.L?1的培养液中常规培养6 h。流式细胞仪测定心肌细胞凋亡率和线粒体膜电位,进行凋亡诱导因子(AIF)的Western blotting分析,检测心肌细胞内SOD活性和MDA含量。结果与缺氧/复氧组相比较,附子多糖后处理可以保护心肌细胞SOD活性,减少MDA的生成,阻止线粒体膜电位的下降,抑制AIF自线粒体向胞浆的释放,减少心肌细胞凋亡率。结论附子多糖后处理对缺氧/复氧后心肌细胞的保护机制与其抗氧化损伤,抑制细胞凋亡的线粒体信号途径有关。     

意大利牛舌草总黄酮及四种化学成分对心肌细胞缺氧/复氧损伤的保护作用及机制研究

目的探讨意大利牛舌草总黄酮及四种化学成分对心肌细胞缺氧/复氧损伤(hypoxia-reoxygenation,H/R)的保护作用及作用机制。方法分离并培养原代乳鼠心肌细胞,随机分组,建立H/R模型,给药后测定相关指标。结果与模型组相比,总黄酮及四种化学成分均能增加心肌细胞缺氧/复氧损伤后的存活率(P<0.01),降低LDH漏出量(P<0.01),减少细胞内MDA的含量(P<0.01),且SOD活力显著性提高(P<0.01),可显著降低心肌细胞早期凋亡指数,提高线粒体膜电位(P<0.01)。给药组NF-κB信号通路关键蛋白的激活受到抑制,Bax/Bcl-2比例明显降低(P<0.01),Beclin-1蛋白表达量明显降低而p62表达上调(P<0.01),焦亡相关蛋白NLRP3、IL-1β以及GSDMD明显降低(P<0.01)。结论意大利牛舌草总黄酮及四种化学成分可通过抑制NF-κB信号通路的激活及心肌细胞的过度自噬,缓解细胞焦亡,减少心肌细胞凋亡,减轻H/R对心肌细胞造成的损伤。     

芳香新塔花对大鼠急性心肌缺血和乳鼠心肌细胞的保护作用

目的比较芳香新塔花Ziziphora clinopodioides不同提取物对大鼠急性心肌缺血、对乳鼠心肌细胞缺氧-复氧损伤的影响。方法通过体内注射垂体后叶素造成大鼠急性心肌缺血模型,观察急性心肌缺血大鼠Ⅱ导联心电图J点的变化,以及试药对超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、丙二醛(MDA)的影响。体外培养原代SD乳鼠心肌细胞,建立心肌细胞缺氧-复氧损伤模型,用MTT法测定药物对心肌细胞存活率的影响,测定给药后MDA的变化。结果芳香新塔花石油醚部位和氯仿部位0.3g/kg显著抑制注射垂体后叶素所致心电图J点升高,对心率、SOD、LDH、MDA的影响不显著;乙醇提取物和氯仿部位能明显降低缺氧-复氧模型下心肌细胞培养液中的MDA浓度。结论芳香新塔花具有潜在的心肌缺血保护作用。     

玉郎伞两种黄酮单体对心肌细胞缺氧/复氧损伤的保护作用

目的研究从玉郎伞(Yulangsan,YLS)中首次分离的两种黄酮单体对体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤的保护作用,并初步探讨其作用机制。方法建立体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤模型,倒置显微镜下观察加入YLS两种单体的含药血清(10%、5%、2.5%)后,各组缺氧/复氧心肌细胞形态学和搏动频率的变化;以MTT法检测各组细胞的存活率;用ELISA法测定心肌细胞Na+,K+-ATP酶、Ca2+,Mg2+-ATP酶活性及细胞培养上清液中总超氧化物歧化酶(T-SOD)、乳酸脱氢酶(LDH)、一氧化氮合酶(NOS)活性和丙二醛(MDA)含量。结果与模型组相比,YLS两种单体含药血清的高、中剂量均能明显增加心肌细胞的存活率,增强Na+,K+-ATP酶、Ca2+,Mg2+-ATP酶活性,降低细胞培养上清液LDH、NOS活性、MDA含量,提高T-SOD活性并呈剂量依赖性(P<0.05)。结论两种YLS单体对体外培养大鼠乳鼠心肌细胞缺氧/复氧损伤具有保护作用,其机制可能与清除自由基、抑制心肌细胞Ca2+超载有关。     

黄芪多糖对乳鼠心肌细胞缺氧/复氧损伤的保护作用

目的探讨黄芪多糖(APS)对乳大鼠培养心肌细胞缺氧/复氧损伤的保护作用及其机制。方法取体外培养的乳大鼠心肌细胞建立缺氧/复氧心肌细胞损伤模型,实验分为5组:空白对照组;模型组(A/R组);3个APS给药组,于缺氧及复氧开始时分别加入终浓度为10、30、100mg·mL-1的APS,观察APS对细胞上清液中缺氧/复氧后LDH、CK、AST漏出量、MDA含量、SOD活性。用MTT法检测APS对缺氧/复氧损伤心肌细胞存活率的影响。用流式细胞仪测定APS对缺氧/复氧损伤心肌细胞凋亡的影响。结果 APS可明显减少缺氧/复氧后LDH、CK、AST漏出量(P<0.05或P<0.01);降低MDA含量,提高SOD活性(P<0.05或P<0.01);明显提高缺氧/复氧损伤心肌细胞的存活率,并能明显抑制缺氧/复氧损伤心肌细胞凋亡。结论 APS对培养心肌细胞的缺氧/复氧损伤具有保护作用,作用机制与其增强细胞抗氧化作用,减少自由基及脂质过氧化物导致的细胞膜损伤和抑制细胞凋亡有关。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

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.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

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.