戈米辛J对肝线粒体膜和培养心肌细胞的抗脂质过氧化作用

戈米辛Ｊ对肝线粒体膜和培养心肌细胞的抗脂质过氧化作用１彭红丽，陈道峰２，蓝洪祥，张雪梅，顾峥，江明华（上海医科大学药学院药理教研室；２生药教研室，上海２０００３２，中国）关键词脂质过氧化；戈米辛Ｊ；木脂素；培养的细胞；肝线粒体；心肌目的：研究来自内南...     

体外细胞三维培养技术在药理毒理学研究中的应用

体外细胞三维培养技术能模拟体内细胞之间以及细胞胞外基质间信号转导的微环境,既能保留体内细胞微环境的物质结构基础,又可体现细胞培养的直观性及条件可控性。近年来,体外细胞三维培养技术在药理毒理学研究中有着广泛应用,是外源化合物遗传毒性、肿瘤多细胞耐药、抗肿瘤药物高通量筛选、皮肤毒理学研究以及药物代谢与毒性试验等研究的有力工具。本文就体外细胞三维培养技术在药理毒理学研究中的应用作简要综述。     

新生大鼠心肌细胞培养技术

目的探讨新生大鼠心肌细胞的分离、培养方法。方法取1~3 d龄新生大鼠的心室肌细胞,用胶原酶Ⅰ分离心肌细胞,离心收集心肌细胞,差速贴壁法纯化后培养于DMEM培养基。显微镜下鉴定心肌细胞的纯度和形态结构,锥虫蓝染色检查心肌细胞成活率。结果心肌细胞纯度为96%,平均成活率95.43%,并出现同簇细胞的同步跳动。结论该方法简单有效,为研究心肌细胞的人员提供了一种实验手段。     

新生大鼠心肌细胞体外原代培养及鉴定

目的探讨一种较为理想的体外原代心肌细胞培养的方法。方法取新生SD大鼠心脏心尖部,冷胰蛋白酶和II型胶原酶．BSA消化分离单细胞,二次差速贴壁法纯化心肌细胞,倒置相差显微镜和流式细胞仪检测细胞存活率、活力及纯度。结果获得的心肌细胞存活率为90％,纯度达94％以上,自发搏动明显,频率为80—120次／min,且培养前15d细胞搏动频率差异无统计学意义（P〉0．05）。结论本方法可获得较高存活率及纯度的心肌细胞,可满足实验要求。     

牛磺酸抗脂质过氧化作用与调节Ca~（2＋）作用

在培养心肌细胞缺氧－再给氧模型上，发现牛磺酸（２０ｍｍｏｌ·Ｌ－１，终浓度）能显著降低细胞内脂质过氧化物（ＬＰＯ）荧光强度；剂量依赖性地提高心肌细胞超氧化物歧化酶（ＳＯＤ）活性；以荧光比率法（荧光探针为Ｉｎｄｏ－１－ＡＭ）在粘附式细胞仪（ＡＣＡＳ５７０）上测定细胞内游离Ｃａ２＋荧光比率，结果显示缺氧及再给氧后细胞内Ｃａ２＋荧光比率明显升高，２０ｍｍｏｌ·Ｌ－１牛磺酸能显著减少Ｃａ２＋荧光比率；分别以２０ｍｍｏｌ·Ｌ－１、０．４ｍｍｏｌ·Ｌ－１Ｃａ２＋及０．１ｍｍｏｌ·Ｌ－１维拉帕米处理细胞，牛磺酸能降低高Ｃａ２＋引起的细胞搏动加快；而提高低Ｃａ２＋及维拉帕米处理后的细胞搏动。提示牛磺酸对培养心肌细胞再给氧损伤有保护作用，其机理与其提高心肌细胞ＳＯＤ活性、降低细胞内游离Ｃａ２＋浓度及对细胞Ｃａ２＋双向调节作用有关。     

丹皮酚抗大鼠心肌缺血再灌注损伤与抗膜脂质过氧化作用

丹皮酚抗大鼠心肌缺血再灌注损伤与抗膜脂质过氧化作用张卫国，张志善（武汉湖北医科大学药理教研室４３００７１）丹皮酚是非甾体类解热镇痛药，有抗动脉粥样硬化斑块形成、抑制血小板聚集（１），及抗心律失常和降压作用（２），并对钙反常培养乳鼠心肌细胞Ｃａ２＋内流...     

血管紧张素Ⅱ对培养乳鼠非心肌细胞DNA及蛋白合成的影响

ＡｕｇⅡ作为一种独立于血液动力学及神经体液因子，对哺乳动物心脏细胞的直接作用尚未完全清楚。本实验中，我们观察了ＡｕｇⅡ对培养心脏的非心肌细胞的作用，结果表明：在培养的非心肌细胞，ＡｕｇⅡ引起蛋白合成、ＤＮＡ合成及细胞数目的明显增加，但不影响细胞的体积。ＡｕｇⅡ在非心肌细胞诱导的ＤＮＡ合成受ＡｕｇⅡ受体拮抗剂［Ｓａｒ１，Ｖａｌ５，Ａｌａ８］－ＡｕｇⅡ完全抑制、这些结果提示：（１）ＡｕｇⅡ诱导心脏非心肌细胞的增殖；（２）ＡｎｇⅡ的上述作用通过ＡＴ１受体亚型调控。     

高山红景天多糖对病毒感染大鼠心肌细胞的抑制作用

目的：观察高山红景天多糖对柯萨奇Ｂ３病毒感染ＳＤ大鼠心肌细胞的抑制作用。方法：采用原代培养乳鼠心肌细胞的方法，建立实验性病毒性心肌炎模型。结果：高山红景天多糖能明显的抑制体外培养心肌细胞在受到柯萨奇Ｂ３病毒感染后导致的心肌酶释放；显著降低病毒在心肌细胞中的增殖量；其半数有效抑制浓度为１５０ｍｇ·Ｌ－１；有效浓度平均抑制率为７１．３％。结论：高山红景天多糖对在ＳＤ大鼠心肌细胞内增殖的柯萨奇Ｂ３病毒具有一定的抑制作用。     

体外全胚胎培养技术及其在药物生殖发育毒性评价中的应用

体外全胚胎培养（whole embryo culture，WEC）技术是一项在受控的体外环境中培养动物胚胎的技术，是研究药物及化学物质在胚胎发育过程中的药理毒理及作用机制的重要方法之一。本文从大鼠和兔的WEC技术发展概况、培养条件、胚胎发育终点的评分系统和拓展的胚胎发育评价终点、药物生殖发育毒理学应用等方面，对WEC技术及近年来研究进展进行介绍。WEC技术是药物生殖发育毒性研究中重要的替代学研究方法之一。其中，对大鼠与兔组合的WEC研究更具有重要价值，可为胚胎发育毒性研究提供更全面的信息。     

高山红景天多糖对病毒感染小鼠心肌细胞的抑制作用

目的：观察高山红景天多糖对柯萨奇Ｂ２病毒感染ＳＤ大鼠心肌细胞的抑制作用，方法：采用原代培养乳鼠心肌细胞的方法，建立实验性病毒性心肌炎模型。结果：高山红景天多糖能明显的抑制休外培养心肌细胞在受到柯萨奇Ｂ３病毒感染后导致的心肌酶释放显著降低病毒在心肌细胞中的增殖量；其半数有效抑制浓度为１５０ｍｇ．Ｌ＾－１，有效浓度平均抑制率为７１．３％，结论：高山红景天多糖对ＳＤ大鼠心肌细胞内增殖的柯萨奇Ｂ３病毒具有     

Polymorphism, pseudopolymorphism, and amorphism of peracetylated alpha-, beta-, and gamma-cyclodextrins

Polymorphism, pseudopolymorphism, and amorphism of hexakis(2,3,6-tri-O-acetyl)-alpha-cyclodextrin (TAalphaCyD), heptakis(2,3,6-tri-O-acetyl)-beta-cyclodextrin (TAbetaCyD), and octakis(2,3,6-tri-O-acetyl)-gamma-cyclodextrin (TAgammaCyD) were investigated using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), powder X-ray diffractometry (XRPD), Fourier transform infrared spectroscopy (FTIR) and optical microscopy. An anhydrous and a bi-hydrate crystalline forms of TAalphaCyD, two monotropic anhydrous polymorphs and three pseudopolymorphs (i.e. methanolate, hydrate, and isopropanolate-hydrate) of TAbetaCyD, as well as two monotropic anhydrous polymorphs and isostructural pseudopolymorphs (e.g. hydrate and isopropanolate-hydrate) of TAgammaCyD were isolated and characterized. The amorphous forms of each TACyD were also obtained. Thermal data for desolvation of TAalphaCyD.2H2O and TAbetaCyD.CH3OH were reconciled with their crystal packing features. Melting temperatures and enthalpies of the crystalline forms of each TACyD can be referred to for possible solid-state interactions with drugs.     

Synthesis,anti-inflammatory,analgesic, and antibacterial activities of some triazole,triazolothiadiazole, and triazolothiadiazine derivatives

This study is concerned with the synthesis of new 1,2,4-triazoles, 1,3,4-thiadiazoles, and 1,3,4-thiadiazines derivatives. Derivatives 3a–i were obtained by condensation of 4-amino-3-(4-pyridine)-5-mercapto-1,2,4-triazole 1 with the appropriate aldehyde. Compounds 4a–i were synthesized in a one pot reaction involving compounds 3a–i, formaldehyde, and morpholine. Condensation of compound 1 with the appropriate acids or 4-substituted phenacyl bromide gave compounds 6a–d and 8a–f respectively. The chemical structures of the newly synthesized derivatives were elucidated using different spectral and elemental methods of analysis. All compounds were evaluated for their anti-inflammatory activity and the most potent derivatives were tested for their analgesic activity using indomethacin as a reference drug. In addition, ulcerogenicity and LD₅₀ for the most active compounds were evaluated. Moreover, the antibacterial activities of the newly synthesized derivatives were investigated.     

New synthesis of benzo-delta-carbolines, cryptolepines, and their salts: in vitro cytotoxic, antiplasmodial, and antitrypanosomal activities of delta-carbolines, benzo-delta-carbolines, and cryptolepines

The paper describes, in its first part, a new synthesis of benzo-delta-carbolines, cryptolepines, and their salts. The strategy is based on the association between halogen-dance and hetero-ring cross-coupling. It is fully convergent and regioselective with interesting overall yields from 27% to 70%. A halogen-dance mechanism in quinoline series is also proposed. The formal synthesis of potential antimalarial compounds and the first total synthesis of 11-isopropylcryptolepine are also described. In the second part, cytotoxic activity against mammalian cells and activities against Plasmodium falciparum and Trypanosoma cruzi of benzo-delta-carbolines and delta-carbolines were evaluated in vitro to study the structure-activity relationships. For benzo-delta-carbolines, methylation at N-5 increases the cytotoxic and antiparasitic activities. A further alkylation on C-11 generally increases the cytotoxic activity but not the antiparasitic activity, cryptolepine and 11-methylcryptolepine being the most active on both parasites. Taking advantage of the fluorescence of the indoloquinoline chromophore, cryptolepine was localized by fluorescence microscopy in parasite DNA-containing structures suggesting that these compounds act through interaction with parasite DNA as proposed for cryptolepine on melanoma cells. For delta-carbolines, methylation at N-1 is essential for the antimalarial activity. 1-Methyl-delta-carboline specifically accumulates in the intracellular parasite. It has weak cytotoxic activity and can be considered as a potential antimalarial compound.     

Stereospecific and circadian-phase-dependent kinetic behavior of d,l-, l-, and d-propranolol in plasma, heart, and brain of light--dark-synchronized rats

In light--dark-synchronized male rats, the kinetic behavior of d,l-, l-, and d-propranolol after single (1.78 and 8.89 mg/kg) or multiple drug administration (6 X 8.89 mg/kg) was studied in plasma, heart, and brain both in the light period (L) and in the dark period (D). With either dosage regimen the kinetics of racemic propranolol displayed a temporal dependency, elimination half-lives in plasma, heart, and brain being shorter during D than during L. This was observed with the stereoisomers only after single drug application with no circadian phase dependency at steady-state concentrations. On the other hand, the kinetic behavior of l- and d-propranolol exhibited pronounced stereospecificity in that t1/2 Beta, Vdbeta, plasma clearance, and drug accumulation in heart and brain were greater for l-propranolol than for the d-isomer. Stereospecific differences in t1/2 beta and elimination rate were more pronounced during D. In the light of the flow-dependent hepatic extraction of propranolol it is unlikely that daily variations in microsomal liver enzyme activity are responsible for the chronopharmacokinetics of propranolol. It is assumed that daily variations in liver blood flow, which is more effectively reduced by beta-receptor blockade in the period of increased sympathetic tone during D, are mainly responsible for the chronopharmocokinetics of the therapeutically used d,l-propranolol.     

PCP, THC, ethanol, and morphine and consumption of palatable solutions

Water-deprived rats were given daily opportunities (2.0-hr sessions) to take water or a sweet solution (20% or 24% sugar-water). After stable intakes of each fluid were achieved, the effects of phencyclidine hydrochloride (PCP), delta-9-tetrahydrocannabinol (THC), ethanol (E), and morphine (M) on intakes were tested. PCP, THC, and M all enhanced intake of the sweet solution, while E produced varying effects across doses tested. With other rats, nearly the same procedure was used except that the test solution presented with water was 0.9% sodium chloride. Doses of PCP enhanced intake of the salty solution. These data, combined with the data from similar studies of the effects of opioids and benzodiazepines, indicate that a wide variety of agents that are self-administered also modify intake of ingesta.     

Combination action of sisomicin, dibekacin and cefotetan, cefotaxime, latamoxef, and cefsulodin against Escherichia coli, Serratia marcescens, and Pseudomonas aeruginosa

The combined actions of sisomicin (SISO), dibekacin (DKB) and cefotetan (CTT), cefotaxime (CTX), latamoxef (LMOX), cefsulodin (CFS) against E coli KC-14, S. marcescens T-55 and P. aeruginosa E-2 were studied. The following results were obtained. The combination of SISO-CTT, SISO-CTX, SISO-LMOX, SISO-CFS, DKB-CTT, DKB-CTX, DKB-LMOX and DKB-CFS using the checker board dilution method on E. coli KC-14, S. marcescens T-55, P. aeruginosa E-2 were found to have a synergistic effect and the minimum FIC index values were 0.26--0.50 for SISO and 0.28--0.75 for DKB, respectively. With the killing kinetic method, all combinations tested showed a synergistic effect.     

Pharmacokinetics of Liquiritigenin in Mice,Rats, Rabbits,and Dogs,and Animal Scale-Up

Pharmacokinetics of liquiritigenin (LQ) and its two glucuronide metabolites, M1 and M2, in mice, rats, rabbits, and dogs and animal scale-up of the pharmacokinetic parameters of LQ were evaluated. After intravenous administration of LQ, the AUC (AUC_0?t) values of LQ, M1, and M2 were proportional to LQ doses in all animals studied. Animal scale-up of some pharmacokinetic parameters of LQ was performed based on the parameters after its intravenous administration (20 mg/kg; in the linear pharmacokinetic range) to the four species. Linear relationships were obtained (r > 0.968) between log CL (or CL/f_u) (L/h) and log species body weight (W) (kg) [CL (or CL/f_u) = 3.29 (34.0) W^{0.723 (0.789)}] and log V_ss (or V_ss/f_u) (L) and log W (kg) [V_ss (or V_ss/f_u) = 0.340 (3.52) W^{0.882 (0.948)}]. Interspecies scale-up of plasma concentration–time data of LQ using apolysichron (complex Dedrick plots) resulted in similar profiles, and plasma concentration–time profile of humans were predicted using the well-fitted four animal data. Our results indicate that the LQ data obtained from laboratory animals could be utilized to generate preliminary estimates of the pharmacokinetic parameters of LQ in humans. These parameters can serve as guidelines for better planning of clinical studies. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4327–4342, 2009     

Comparison of natriuretic, uricosuric, and antihypertensive properties of tienilic acid, bendrofluazide, and spironolactone.

The antihypertensive properties of the new diuretic tienilic acid were investigated. Thirteen previously untreated hypertensive patients took part in a double-blind crossover study in which 30 days' treatment with tienilic acid 250 mg, bendrofluazide 5 mg, and spironolactone 100 mg were compared. Bendrofluazide caused the greatest natriuresis on the first treatment day and the most rapid fall in blood pressure. The ultimate antihypertensive effect of all three drugs was similar. Tienilic acid caused a noticeable reduction in serum urate concentrations and a rise in urate clearance, in contrast to the other two agents, which caused slight urate retention. Tienilic acid and bendrofluazide caused falls and spironolactone a rise in plasma potassium concentrations. No untoward effects were seen from any of the drugs. It is concluded that tienilic acid is a moderately potent diuretic that lowers plasma urate concentrations. It may be the drug of first choice for hypertensive patients who already have gout or are likely to develop it when taking thiazide diuretics.     

Pharmacokinetics, tissue distribution, and excretion of sitafloxacin, a new fluoroquinolone antibiotic, in rats, dogs, and monkeys

The pharmacokinetics, tissue distribution and excretion of sitafloxacin (CAS 127254-12-0, DU-6859a) were investigated in rats, dogs, and monkeys following single intravenous or single oral administration of 14C-labelled sitafloxacin at a dose of 4.69 mg/kg. Following single administration of the oral dose, serum concentrations of radioactivity peaked at 0.5 h in rats, 2.3 h in dogs, and 2.5 h in monkeys. The apparent absorption ratios of 14C-sitafloxacin based on the AUC0-infinity were 31%, 51%, and 93% in rats, dogs, and monkeys, respectively. In rats, the drug-related radioactivity had been distributed to most organs and tissues 30 min after oral dosing, and had been essentially eliminated after 24 h. The highest levels of radioactivity were observed in the kidneys and liver, whereas the concentrations in the cerebrum and spinal cord were much lower than the serum value. The urinary recoveries of radioactivity after intravenous dosing were 45.5 % in rats, 32.3 % in dogs, and 77.8 % in monkeys. In bile duct-cannulated rats, 57.8 % of the orally administered radioactivity was excreted in the bile within 48 h, and at least 45 % of the sitafloxacin-related material secreted in the bile was re-absorbed from the gastrointestinal tract. These results indicate that sitafloxacin is rapidly absorbed and widely distributed into various tissues. Sitafloxacin-related material is eliminated primarily through both renal and biliary excretion in rats, and possibly in dogs, whereas renal excretion is the major route of elimination in monkeys.     

Storage,release, uptake,and inactivation of purines

Adenine nucleotides are released into the interstitial space during platelet thrombus formation and neurotransmission. ATP has also been reported to be released from the heart and endothelial cells in some studies. Ecto ATPase, ADPase, and 5′-nucleotidase activities capable of hydrolyzing ATP sequentially to adenosine are present in many cell types and may serve to terminate the actions of the nucleotides. The opposing effects of adenosine and ATP on the same cell types have suggested a modulatory role for adenosine of the actions of extracellular ATP and that the rates of hydrolysis of nucleotides might be regulated. Consistent with this it has been found that the balance between feedforward inhibition of 5′-nucleotidase by ADP and/or ATP and preferential delivery of AMP from ADPase to 5′-nucleotidase determines the rate of adenosine production and that this differs in different cell types. Alternatively, adenosine may be produced intracellularly as a result of an imbalance between energy demand and supply. There are at least two different cytosolic forms of 5′-nucleotidase. Degradation of ATP during increased metabolic activity results in an increase in intracellular AMP concentration. Either cytosolic enzyme has a high KM (2–5 mM) and would thus respond to this increase with a proportional rise in the rate of adenosine production. The nucleoside transporter is essential to allow the diffusion of adenosine to extracellular receptor sites. In general, adenosine must be taken up via the nucleoside transporter before it is inactivated either by phosphorylation by adenosine kinase in the micromolar range or by deamination by adenosine deaminase at higher concentrations. © 1993 Wiley-Liss, Inc.