不同剂量染料木黄酮及其代谢产物在大鼠胆汁中的排泄动力学

目的研究不同剂量染料木黄酮及其葡糖醛酸化代谢产物在大鼠胆汁内的排泄动力学。方法将染料木黄酮制成混悬液，分别按6.25，12.5和50 mg·kg^-1给大鼠灌胃，于灌胃后不同时间收集胆汁，用葡糖醛酸酶溶液处理胆汁。采用高效液相色谱法测定胆汁中染料木黄酮及其葡糖醛酸化代谢产物的浓度。结果3种剂量6.25，12.5和50 mg·kg^-1分别给药后，累积以原形从胆汁排泄的药物分别为(42.56±6.54)，(75.17±18.87)和(126.60±34.78) μg，累积经胆汁排泄的总药物(原形药物+葡糖醛酸化药物)分别为(108.46±35.23)，(423.46±158.31)和(853.74±320.84) μg，葡糖醛酸化代谢产物分别占胆汁排泄总量(原形药物+葡糖醛酸化药物)的60.76%，82.25%和85.17%。结论染料木黄酮在大鼠胆汁中主要以葡糖醛酸结合形式排泄，原形药物及其葡糖醛酸化代谢产物在大鼠胆汁中的排泄呈现明显的非线性剂量依赖性特征。     

LC/MSn鉴定咖啡酸在大鼠体内的代谢产物

目的 研究咖啡酸（CA）在大鼠体内的代谢产物。方法 大鼠灌胃（50 mg·kg-1）给予CA后采集0 ~ 4 h尿样,用电喷雾离子阱多级质谱法对CA在大鼠体内的代谢产物进行分析。结果 大鼠灌胃给予CA后,在体内可测到2个原形药的甲基化代谢物、2个原形药的单葡萄糖醛酸结合物、1个原形药的双葡萄糖醛酸结合物、2个原形药的单硫酸结合物、2个甲基化物的葡萄糖醛酸结合物和2个甲基化物的硫酸结合物。结论 CA在大鼠体内广泛代谢,其代谢物的结构有待于进一步分析后确证。     

HPLC-ESI/MS分析5-羟基黄酮在大鼠体内的代谢产物

目的研究5-羟基黄酮在大鼠体内的代谢产物。方法应用高效液相色谱-电喷雾质谱(HPLC-ESI/MS)检测大鼠灌胃5-羟基黄酮后血浆、尿液、胆汁和粪便中的代谢产物。实验采用Zorbax C18色谱柱,0.05%甲酸乙腈-0.05%甲酸水二元线性梯度洗脱进行色谱分离,并与电喷雾质谱联用,根据正离子模式的分子离子峰获得化合物分子量信息,推测化合物的可能结构。结果仅在大鼠粪便中检测到原形成分,在大鼠尿液、粪便、血浆、胆汁中检测到5-羟基黄酮葡糖醛酸结合物。结论 5-羟基黄酮吸收差,在大鼠体内主要以II相代谢产物葡萄糖醛酸结合物的形式存在。     

曲克芦丁及其代谢产物在大鼠体内的排泄动力学研究

目的研究曲克芦丁及其代谢产物在大鼠体内的排泄情况。方法大鼠腹腔注射曲克芦丁,收集尿液、粪便和胆汁样品,应用 HPLC方法测定曲克芦丁在大鼠尿液、粪便和胆汁样品的含量,测定曲克芦丁代谢产物在大鼠粪便中的含量。结果曲克芦丁在大鼠尿液、粪便和胆汁中的累积排泄率分别为( 16.17±10.28)%、(0.54±0.47)%和( 58.94±13.37)%。与此同时,粪便中约有( 22.69±12.48)%的代谢产物曲克芦丁苷元生成。结论曲克芦丁主要通过胆汁进行排泄,以原型和代谢产物的形式排出体外。     

高效液相-电喷雾质谱分析7-羟基黄酮在大鼠体内的代谢产物

目的:研究7-羟基黄酮在大鼠体内的代谢。方法:应用高效液相-电喷雾质谱检测大鼠灌胃7-羟基黄酮后血浆、尿液、胆汁和粪便中的代谢产物。实验采用Zorbax C18色谱柱,二元线性梯度洗脱进行色谱分离,并与电喷雾质谱联用,根据负离子模式的分子离子峰获得化合物相对分子质量信息,推测化合物的可能结构。结果:在大鼠尿液、粪便、血浆、胆汁中检测到原形成分7-羟基黄酮和7-羟基黄酮葡萄糖醛酸结合物,在胆汁或尿中尚检测到7-羟基黄酮硫酸结合物。结论:7-羟基黄酮在大鼠体内主要以Ⅱ相代谢产物葡萄糖醛酸结合物和硫酸结合物的形式存在。     

HPLC法测定大鼠尿液中瑞香素的浓度

目的:建立简单、灵敏、准确的大鼠尿液中瑞香素浓度的HPLC测定方法。方法:尿样经乙酸乙酯萃取,采用HPLC法检测,色谱柱为Agilent Zorbax SB-C18(250 mm×4.6 mm,5μm)柱,流动相为甲醇-0.5%醋酸水(22∶78),流速1 mL·min-1,检测波长327 nm。大鼠灌胃不同剂量的瑞香素固体分散体溶液(5,25,125 mg·kg-1),收集0～2,2～6,6～12,12～24,24～48 h的尿液并分析,计算瑞香素在尿液中的累积排泄率。结果:瑞香素尿液的线性范围为0.155～396μg.mL-1(r=0.9991),定量限为0.155μg·mL-1。大鼠灌胃给药瑞香素后2 h即能检测到原型药物,同时发现其代谢产物,24 h内经尿液排泄基本完全,累积排泄率不到6%。结论:所建立的方法准确可靠,可用于大鼠灌胃瑞香素后原型药物尿样浓度的测定及其尿排泄研究。     

LC/MSn鉴定咖啡酸在大鼠体内的代谢产物

目的研究咖啡酸（CA）在大鼠体内的代谢产物。方法大鼠灌胃（50mg·kg^-1）给予CA后采集0～4h尿样,用电喷雾离子阱多级质谱法对CA在大鼠体内的代谢产物进行分析。结果大鼠灌胃给予CA后,在体内可测到2个原形药的甲基化代谢物、2个原形药的单葡萄糖醛酸结合物、1个原形药的双葡萄糖醛酸结合物、2个原形药的单硫酸结合物、2个甲基化物的葡萄糖醛酸结合物和2个甲基化物的硫酸结合物。结论CA在大鼠体内广泛代谢,其代谢物的结构有待于进一步分析后确证。     

牡荆素在大鼠体内的代谢研究

目的 研究黄酮碳苷牡荆素在大鼠体内的代谢产物,并推测其代谢途径。方法 SD大鼠灌胃给予5 mg·kg^-1牡荆素,收集0~3 h,3~6 h,6~12 h的尿液,采用UPLC-Q-TOF检测尿样中代谢产物。结果 采用Metabolynx XS代谢物分析软件,根据质谱碎片信息对代谢物进行结构鉴定,最终得到3个代谢产物。结论 牡荆素在大鼠尿液中检测得到1个一相代谢产物,2个二相代谢产物,推测牡荆素在大鼠体内主要发生氧化、甲基化和葡萄糖醛酸结合反应,其中葡萄糖醛酸化反应是较强的代谢种类。     

液相色谱-串联质谱法分析抗肿瘤新药乙烷硒啉在大鼠体内的代谢产物

本研究对抗肿瘤新药1,2-[二(1,2-苯并异硒唑-3(2H)-酮)]乙烷(乙烷硒啉,BBSKE)在大鼠体内的代谢产物进行鉴定。在灌胃给予大鼠单剂量乙烷硒啉200mg·kg-1后,采用液相色谱-串联质谱法(LC-MSn)对大鼠尿液、粪样、胆汁和血浆中的代谢产物进行检测,通过全扫描和选择离子扫描,以及根据多级质谱裂解规律对代谢物的结构进行分析。研究发现在大鼠尿样、粪样、胆汁和血浆中检测到3种Ⅰ相代谢产物和1种Ⅱ相代谢产物,其代谢途径分别为氧化、甲基化、硫甲基化和葡萄糖醛酸化反应,提示乙烷硒啉在大鼠体内的代谢方式可能是通过氧化、甲基化及葡萄糖醛酸化反应形成代谢产物。     

染料木黄酮在雌雄大鼠肝微粒体中的代谢差异

目的研究染料木黄酮在♀、♂大鼠肝微粒体中的代谢差异。方法制备♀、♂大鼠肝微粒体,确定染料木黄酮代谢的酶动力学条件,分别用CYP1A2抗体和选择性CYP1A2抑制剂呋喃茶碱与大鼠肝微粒体和染料木黄酮共同温孵,测定染料木黄酮在♀、♂大鼠肝微粒体中的代谢速率,评价♀、♂大鼠CYP1A2的相对百分比活性。结果在CYP1A2抗体浓度为1∶400,孵育时间为30 m in条件下,♂大鼠肝微粒体代谢染料木黄酮的相对代谢率为(20.95±2.13)%,♀动物为(13.73±1.26)%。在选择性CYP1A2抑制剂呋喃茶碱浓度为3.125μmol.L-1,孵育时间为30 m in条件下,♂动物为(58.02±3.35)%,而♀大鼠肝微粒体代谢染料木黄酮的相对代谢率为(43.82±2.65)%,两者之间差异有显著性(P<0.01)。结论染料木黄酮在♂大鼠肝微粒体中代谢较♀大鼠快,提示♂大鼠肝微粒体CYP1A2酶活性高于♀大鼠。     

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.

     

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.     

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.     

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.