脑钠肽对外周血内皮祖细胞增殖、迁移与黏附能力的影响

目的研究脑钠肽对外周血内皮祖细胞数量及增殖、迁移、黏附能力的影响。方法选取30例健康成人,取其外周静脉血,采用密度梯度离心法分离外周血获得单个核细胞,将其接种在人纤维连接蛋白包被的培养板上,经血管内皮生长因子（Vascular endothelial growth factor,VEGF）诱导培养,7d后获得贴壁细胞,荧光显微镜鉴定FITC-UEA-I和Dil-acLDL双染色为正在分化的内皮祖细胞（endothelial progenitor cells,EPCs）。收集EPCs随机分成6组,分别为对照组和不同浓度脑钠肽干预组（0.05μg/ml、0.10μg/ml、0.15μg/ml、0.20μg/ml、0.25μg/ml）,均培养24h后采用MTT比色法测定内皮祖细胞（EPCs）的增殖能力,改良的Boyden小室法测定EPCs的迁移能力,黏附实验测定内皮祖细胞的黏附能力。结果不同浓度的重组人脑钠肽干预组均显著增加EPCs的增殖及迁移能力,分别与两者呈一定的量效关系,对黏附能力无明显影响。结论脑钠肽能促进外周血内皮祖细胞增殖、迁移能力,对内皮祖细胞的黏附能力无明显影响。     

U937泡沫细胞中血管内皮生长因子的表达及药物的抑制作用

目的研究U937泡沫细胞中血管内皮生长因子(VEGF)表达以及丹酚酸B和银杏叶提取物对其的抑制作用。方法U937细胞与80 mg·L^-1氧化型低密度脂蛋白(OX-LDL)孵育48 h,建立U937泡沫细胞模型。用基础酶联免疫吸附试验(ELISA)检测各组培养基中VEGF蛋白分泌含量,用免疫组织化学法检测VEGF蛋白表达,用原位杂交法检测VEGF mRNA水平。给予不同浓度丹酚酸B和银杏叶提取物,观察VEGF表达的变化。结果培养的U937泡沫细胞培养液中VEGF含量和细胞中VEGF表达均明显高于U937正常细胞。丹酚酸B和银杏叶提取物加入后阳性反应明显减弱。结论泡沫细胞中VEGF呈高表达、含量明显增加,而丹酚酸B和银杏叶提取物可明显降低其含量。     

VEGF诱导牛主动脉内皮细胞通透性升高及丹酚酸B对其的抑制作用(英文)

目的:考察血管内皮生长因子/血管通透因子(VEGF)对体外培养牛主动脉内皮细胞脂蛋白通透性的影响及丹酚酸B的抑制作用。方法:将不同浓度的VEG、丹酚酸B及培养基加入牛主动脉内皮细胞单层,与~(125)I-LDL共孵育,在不同的时间段用SN-695型液闪计数器测其通过细胞单层的百分率。结果:VEGF可显著增强牛主动脉内皮细胞单层对~(125)I-LDL的通透性(P<0.01),这种增加具有时间和剂量依赖性。丹酚酸B能显著抑制VEGF诱导的这种高通透(P<0.01)。结论:上述结果提示VEGF在动脉粥样硬化的形成和发展过程中起一定作用。丹酚酸B对VEGF诱导血管内皮通透性升高有显著的抑制作用。     

丹酚酸B对鸡胚绒毛尿囊膜血管新生作用的影响

目的探讨丹酚酸B对鸡胚绒毛尿囊膜血管新生的影响。方法将7日龄鸡胚制备鸡胚绒毛尿囊膜(chick embryo chorioallantoic membrane,CAM)模型,随机分为6个组,分别为PBS(phosphatebuffer solution)对照组,丹酚酸B 4个剂量组(150、50、16.67、5.56 mg.L-1)及VEGF(vascular endo-thelial growth factor)组,培养3 d后时采集数据进行评价。结果丹酚酸B 150、50 mg.L-1组血管新生面积明显高于PBS(phosphate buffer solution)对照组(P<0.01,P<0.05)。结论丹酚酸B可明显促进鸡胚绒毛尿囊膜血管新生。     

SDF 1α对大鼠骨髓源性内皮祖细胞克隆形成能力及增殖的影响

目的观察基质细胞衍生因子-1α（SDF-1α）对体外培养的大鼠骨髓源性内皮祖细胞（EPCs）动员和增殖的影响。方法微孔法获取大鼠骨髓EPCs并采用荧光显微镜和流式细胞术鉴定EPCs特异性标记物。不同浓度SDF-1α处理EPCs后,采用细胞培养、MTT检测EPCs的克隆形成、细胞增殖。结果通过MTT法检测,对照组与（1、10、100μg/L）SDF-1α处理组的OD值分别为0.311±0.054、0.587±0.072、0.813±0.056、1.029±0.078,通过统计学方法分析,对照组与SDF-1α处理组比较差异均有统计学意义（n=5,P〈0.05）;100μg/LSDF-1α处理组次级内皮祖细胞集落单位数目是对照组近3倍（4.67±1.577比14.33±3.055,n=5,P〈0.01）。结论SDF-1α剂量依赖性地促进EPCs增殖,并显著增强EPCs克隆形成能力。     

缺氧对血管内皮生长因子诱导牛冠状动脉内皮细胞脂蛋白通透性升高的调控及丹酚酸B的抑制作用

目的　为了了解缺氧对血管内皮生长因子(VEGF)增强血管内皮细胞通透性作用的影响及其与动脉粥样硬化的关系 ,考察了正常和缺氧状态下VEGF对体外培养牛冠状动脉内皮细胞 (BCEC)脂蛋白通透性的影响及丹酚酸B的抑制作用。方法　正常及缺氧条件下 ,将VEGF及丹酚酸B加入BCEC共孵育 ,用SN 695型液闪计数器测 [12 5I]低密度脂蛋白([12 5I]LDL)通过BCEC的百分率。结果　VEGF可显著增强BCEC对 [12 5I]LDL的通透性 ,这种增加具有浓度依赖性。缺氧 3h可促进VEGF所致的的通透性增加。丹酚酸B在正常和缺氧条件下均显著抑制VEGF诱导的BCEC通透性升高。结论　VEGF可增强血管内皮细胞的通透性 ,可能在动脉粥样硬化的形成和发展过程中起一定作用。丹酚酸B对VEGF诱导的血管内皮通透性升高有显著的抑制作用     

丹酚酸B通过抑制NLRP3炎症小体priming阶段减轻缺氧诱导大鼠心肌细胞损伤

目的探究丹酚酸B(Sal B)能否通过调控NLRP3炎症小体priming阶段减轻缺氧诱导大鼠心肌细胞损伤。方法CCK8法检测不同浓度丹酚酸B对大鼠H9C2细胞生长的影响,并挑选出合适的丹酚酸B实验浓度;微孔板、ELISA法检测缺氧造模后实验组,不同浓度给药组,以及抑制剂组LDH/cTn/IL-1β的分泌水平;qPCR以及蛋白质免疫印迹法检测造模后实验组,不同浓度给药组以及抑制剂组TLR4/Myd88/IRAK1/NF-κB/NLRP3基因以及蛋白表达水平。结果经过缺氧处理后,H9C2细胞活性下降,LDH/cTn/IL-1β分泌量升高,TLR4/Myd88/IRAK1/NF-κB/NLRP3在mRNA水平以及蛋白表达上调;与模型组相比,丹酚酸B预处理24h后,H9C2细胞活性增加,LDH/cTn/IL-1β分泌量下降,TLR4/Myd88/IRAK1/NF-κB/NLRP3 mRNA水平以及蛋白表达水平降低。结论丹酚酸B可以通过调控NLRP3炎症小体priming阶段,减轻缺氧诱导的大鼠心肌细胞损伤。     

阿霉素与顺铂联合重组人血管内皮抑素对骨肉瘤患者血清血管内皮生长因子和基质金属蛋白酶9水平的影响

目的 观察阿霉素与顺铂联合重组人血管内皮抑素对骨肉瘤患者血清血管内皮生长因子(VEGF)和基质金属蛋白酶(MMP)-9水平的影响.方法 42例骨肉瘤患者完全随机分为研究组19例和对照组23例,对照组给予阿霉素25 mg/m静脉注射,第1～3天;顺铂100 mg/m2(静脉滴注24h);研究组给予重组人血管内皮抑素7.5 mg/m2静脉滴注,第1 ～ 14天;2组均21 d为1个周期,2个周期为1个疗程.治疗前后检测2组患者血清VEGF及MMP-9的水平.结果 治疗后研究组血清VEGF及MMP-9表达水平明显低于对照组[(356±282)μg/L比(518 ±503) μg/L,(213±192) μg/L比(382±343)μg/L,均P＜0.05].结论 阿霉素与顺铂联合重组人血管内皮抑素能够明显降低骨肉瘤患者外周血VEGF及MMP-9的表达水平.     

双氯酚酸钠抑制VEGF对角膜新生血管调控作用的研究

目的探讨双氯酚酸钠抑制血管内皮生长因子(VEGF)对调控角膜新生血管的作用。方法建立化学伤后大鼠角膜新生血管动物模型,随机分为双氯酚酸钠组和生理盐水组。计算两组各个阶段新生血管面积,免疫组织化学染色方法检测VEGF蛋白在角膜各层中的分布,测定积分光密度确定表达的蛋白量。结果正常SD大鼠角膜中表达极低的VEGF蛋白;各个时段实验组的血管生长速度和面积均小于对照组(P<0.05);各个时段实验组VEGF表达少于对照组(P<0.01),均具有显著差异;各组内VEGF表达和角膜新生血管生成具有显著正相关(P<0.05)。结论双氯酚酸钠可以抑制碱烧伤后角膜VEGF的表达,从而可以调控角膜修复和炎症性新生血管生成过程。     

丹酚酸B对自发性糖尿病大鼠合并大血管病变的保护作用

摘 要 目的：探讨丹酚酸B对自发性糖尿病（GK）大鼠合并大血管病变的保护作用及可能的机制。方法: 将32只GK大鼠随机分为模型组和丹酚酸B低、中、高剂量组(40,80,160 mg·kg^-1),另选取正常8只Wistar大鼠作为正常对照组。所有GK大鼠给予高糖、高脂饲料16周后以L-N-硝基甲酯（L-NAME）10 mg·kg^-1·d^-1灌胃,连续8周。给药开始及结束前对大鼠的血糖和血压进行检测,麻醉后取腹主动脉血进行血常规检测,放血后取胸主动脉制作病理切片,观察血管病变,Western-blot法检测血管内皮细胞生长因子（VEGF）的表达。结果: 所有GK大鼠血糖水平持续升高,但模型组与给药组差异无统计学意义（P>0.05）,模型组大鼠实验结束时血压显著升高,给予丹酚酸B可剂量依赖性降低血压,差异有统计学意义（P＜0.05）;丹酚酸B可显著降低模型大鼠的白细胞总数和分类计数,明显减轻血管病变程度并减少VEGF的表达。结论:丹酚酸B对自发性糖尿病大鼠的大血管具有明显的保护作用,其可能的机制为降低血压、抗炎以及降低VEGF的表达。     

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.