昆布提取物J201对博莱霉素诱导的大鼠肺纤维化的保护作用研究

目的：研究昆布提取物J201对大鼠肺纤维化的治疗作用及其可能的作用机制。方法：Wistar大鼠64只，随机分为4组。气管内灌注博莱霉素（BLM）诱导肺纤维化模型，观察BLM诱导的大鼠肺纤维化经不同剂量的J201治疗后，肺组织形态学、肺系数、丙二醛（MDA）和羟脯氨酸（HYP）含量的变化以及Fas、Fas配体（FasL）、基质金属蛋白酶-9（MMP-9）、组织金属蛋白酶抑制剂-2（TIMP-2）表达的变化。结果：J201能明显改善BLM诱导的大鼠肺纤维化肺组织形态学改变，并呈现一定的量效关系；能显著降低肺系数、肺组织MDA和HYP的含量，对肺组织Fas、FasL、MMP-9、TIMP-2的表达有明显的抑制作用，对肺系数、MDA、HYP、Fas、FasL、MMP-9 TIMP-2的影响呈现一定的量效关系。结论：J201对实验性大鼠肺纤维化有一定的保护作用，抗氧化作用及抑制Fas、FasL、MMP-9、TIMP-2的表达是其可能的机制之一。     

The effect of a liver-X-receptor ligand on bleomycin induced pulmonary fibrosis in mice

The liver-X-receptors have shown anti-fibrosis ability in several animal models. Our purpose was to investigate the effect of LXRs in bleomycin induced lung fibrosis in mice. Bleomycin was intratracheally delivered to mice. Some mice were administered a LXR agonist, T0901317. Then mice were evaluated for the development of lung inflammation and fibrosis. T0901317 was able to attenuate the inflammation and fibrosis induced by bleomycin. T0901317 treatment evidently abolished the high level of TGF-β1 and inhibited NF-κB DNA-binding activity in lung. So LXRs may attenuate the progressing of lung fibrosis, providing a potential treatment of IPF.     

In vitro prebiotic effects and quantitative analysis of Bulnesia sarmienti extract

Prebiotics are used to influence the growth, colonization, survival, and activity of probiotics, and enhance the innate immunity, thus improving the health status of the host. The survival, growth, and activity of probiotics are often interfered with by intrinsic factors and indigenous microbes in the gastrointestinal tract. In this study, Bulnesia sarmienti aqueous extract (BSAE) was evaluated for the growth-promoting activity of different strains of Lactobacillus acidophilus, and a simple, precise, cost-effective high-performance liquid chromatography (HPLC) method was developed and validated for the determination of active prebiotic ingredients in the extract. Different strains of L. acidophilus (probiotic) were incubated in de Man, Rogosa, and Sharpe (MRS) medium with the supplementation of BSAE in a final concentration of 0.0%, 1.0%, and 3.0% (w/v) as the sole carbon source. Growth of the probiotics was determined by measuring the pH changes and colony-forming units (CFU/mL) using the microdilution method for a period of 24 hours. The HPLC method was designed by optimizing mobile-phase composition, flow rate, column temperature, and detection wavelength. The method was validated according to the requirements of a new method, including accuracy, precision, linearity, limit of detection, limit of quantitation, and specificity. The major prebiotic active ingredients in BSAE were determined using the validated HPLC method. The rapid growth rate of different strains of L. acidophilus was observed in growth media with BSAE, whereas the decline of pH values of cultures varied in different strains of probiotics depending on the time of culture. (+)-Catechin and (−)-epicatechin were identified on the basis of their retention time, absorbance spectrum, and mass spectrometry fragmentation pattern. The developed method met the limit of all validation parameters. The prebiotic active components, (+)-catechin and (−)-epicatechin, were quantified as 1.27% and 0.71% (w/w), respectively, in crude extract, and 6.36 ± 0.06 μg/ mL and 4.47 ± 0.41 μg/mL (mean ± standard deviation), respectively, in a prebiotic capsule of BSAE by HPLC analysis. BSAE contains the active components of prebiotics and enhances the growth of L. acidophilus.     

Respirable dry powder formulation of bleomycin for developing a pulmonary fibrosis animal model

The main purpose of the present study was to develop a respirable powder (RP) formulation of bleomycin (BLM) as a research tool for developing a pulmonary fibrosis animal model. The BLM-RP was prepared with a jet-milling system, the physicochemical properties of which were characterized focusing on morphology, stability, particle size distribution, and inhalation performance. Under an accelerated condition, the BLM-RP was superior to BLM solution in terms of its stability. Cascade impactor analyses demonstrated high inhalation performance with emitted dose and fine particle fraction of approximately 99% and 46%, respectively. Intratracheal administration of the BLM-RP (3 mg BLM/kg) in rats led to significant increases in collagen production and recruitment of inflammatory cells in lung by approximately 1.5- and 29-fold, respectively. The collagen overexpression was consistent with the results from picrosirius red staining of lung tissues in the rats treated with BLM-RP. Inhaled tranilast (TL; 100 μg/rat), an antifibrotic agent, could ameliorate inflammatory/fibrotic responses with reductions of recruited inflammatory cells and collagen content by 32% and 59%, respectively, validating the pulmonary fibrosis animal model. From these findings, the BLM-RP with improved stability could be a beneficial research tool for developing a pulmonary fibrosis model in drug discovery for antifibrotic drug candidates.     

灯盏花素对实验性肺纤维化大鼠的干预作用

目的探讨灯盏花素对博莱霉素致肺纤维化大鼠肺组织细胞凋亡、Fas/FasL表达以及氧自由基的影响。方法48只大鼠随机分为对照组(16只)、模型组(16只)以及灯盏花素组(16只),气管内注入博莱霉素复制大鼠肺纤维化模型,次日灯盏花素组每日腹腔内注射灯盏花素注射液10mg.kg-1,于d7及d28每组分别处死8只大鼠,运用流式细胞仪测定肺纤维化大鼠肺组织细胞凋亡率、Fas/FasL表达,用TAB法测定肺组织匀浆丙二醛含量以及肺组织匀浆羟脯氨酸含量。结果炎症期和纤维化期大鼠肺组织细胞凋亡率增加,Fas/FasL表达增多,丙二醛及羟脯氨酸含量增高(P<0.01),而灯盏花素能明显降低肺组织的凋亡率,降低丙二醛及羟脯氨酸含量,下调Fas/Fasl的阳性表达。结论灯盏花素能减慢肺间质纤维化的进程,对肺间质纤维化有一定的保护作用。     

Comparative pharmacokinetics of vitamin A (retinol) in rats and hamsters

The absorption and elimination of all-trans-retinol in the plasma of rats and hamsters were studied after an oral dose of 45 mg/kg body weight. The hamsters exhibited a higher pretreatment mean circulating retinol concentration than rats maintained on an identical diet. The increase in plasma retinol after a single oral dose was much greater in hamsters than rats. The area under the plasma concentration v. time curve was approximately 60% greater for rats than for hamsters. The elimination half-times for rats were much longer than for hamsters. Plasma retinol uptake and disappearance were consistent with a two-compartment open model with first-order absorption.     

A nutraceutical extract from Inula viscosa leaves: UHPLC-HR-MS/MS based polyphenol profile,and antioxidant and cytotoxic activities

Nowadays, advanced extraction techniques and highly sensitive metabolic profiling methods are effectively employed to get new information on plant chemical constituents. Among them wild medicinal plants or their parts, with large and ancient use in folk medicine, are investigated for their potential functional use and cultivation. In this context, Inula viscosa leaves engaged our attention. A simple experimental design, based on Soxhlet extraction and chromatographic fractionation, allowed us to obtain the investigated polyphenol fraction (IvE). UHPLC-HRMS analyses revealed shikimoyl depsides of caffeic acid and unusual dihydrobenzofuran lignans as main secondary metabolites. These compounds, together with cinchonain-type phenols, and hydroxycinnamoyl flavonol glycosides, are reported for the first time in inula. Overall, forty-three secondary metabolites were identified. The extract exerted a remarkable antiradical activity towards DPPH^• and ABTS^+•. Furthermore, it was able to inhibit cell viability and mitochondrial redox activity of neuroblastoma, hepatoblastoma and colon carcinoma cells, whereas it did not affect cell density of HaCaT cells immortalized human keratinocytes. As detected by the oxidant-sensing probe 2′,7′-dichlorodihydrofluorescein diacetate, the inhibitory responses seemed to be related to IvE-induced increase of intracellular reactive oxygen species (ROS). The obtained results highlighted that inula leaves, nowadays even undervalued and unexplored, could be considered a renewable source of nutraceutical compounds.     

高效液相色谱法测定人血浆中尼莫地平的浓度

目的建立人血浆中尼莫地平浓度的高效液相色谱测定方法.方法血样碱化后用乙醚萃取,在Hypersil-BDS色谱柱上,以乙腈-水(70∶30)为流动相进行色谱分离,检测波长为 238 nm.结果尼莫地平浓度在1.96 ～196.0 ng/mL范围线性相关(r=0.9997,n=5).最低检测限约为0.98 ng/mL.三种浓度的相对回收率为98.56%～102.9%,日内RSD为2.82%～4.33%(n=5), 日间RSD为4.74%～6.05%(n=5).结论本法简便、快速,适合药代动力学的研究.     

High-Performance Liquid Chromatography Determination of Acitretin in Plasma and Its Application to a Pharmacokinetic Study in Human Subjects

Pharmaceutical Research -     

Fast separation of microcystins and nodularins on narrow-bore reversed-phase columns coupled to a conventional HPLC system

Six modern narrow-bore (50 mm length × 2-2.1 mm internal diameter) reversed-phase HPLC columns were tested in the rapid separation of ten microcystins and three nodularins, cyanobacterial peptide hepatotoxins. The columns had either a sub-3 μm particle size or were of monolithic silica technology for high efficiency and rapid run times. A standard low-pressure gradient HPLC equipment with diode-array UV detector was used for the chromatography. The gradient mobile phase consisted of water and acetonitrile, both acidified with trifluoroacetic acid, and the gradient rise times were 1-4 min. Chromatographic performance was assessed by visual judgement and by calculating parameters such as capacity factors, resolution, peak width, selectivity and peak form. Several chromatographic conditions produced excellent results. We recommend sub-3 μm particle-based or monolithic reversed-phase columns for routine use in microcystin and nodularin HPLC analyses.     

盐酸山莨菪碱对头孢拉定注射液稳定性的影响

目的:研究盐酸山莨菪碱(654-2)对头孢拉定(Cep)注射液稳定性的影响.方法:采用反相高效液相色谱法测定3种温度F,不同时间的Cep在生理盐水和含654-2盐水中的含量.结果:两组数据经统计学处理无显著性差异.结论:Cep与654-2配伍时其含量稳定.     

去铁酮在大鼠体内的药代动力学与组织分布

目的研究去铁酮(DFP)在大鼠体内的药代动力学和组织分布。方法雄性Wistar大鼠ig给予DFP35,70和140mg.kg-1后,于不同时间点收集血液和组织样本。采用高效液相色谱法测定大鼠血浆及组织中的DFP的含量,运用DAS2.0药代动力学智能分析软件拟合房室模型,并进行药代动力学参数计算。结果大鼠ig给予DFP35,70和140mg.kg-1后,体内药代动力学过程符合二室模型,t1/2α分别为23.3,22.2和20.9min,t1/2β分别为53.3,50.9和46.3min,Cl分别为0.017,0.021和0.016L.min-1.kg-1。大鼠ig给予DFP70mg.kg-1后,DFP在胃和肝中浓度较高,60min时肝中DFP含量可达(359.22±31.16)μg.g-1,其他组织含量较低。结论DFP在大鼠体内吸收和消除较迅速,在体内组织分布广。     

克林霉素与酚磺乙胺的配伍稳定性研究

目的在室温下6 h内考察克林霉素与酚磺乙胺的配伍及稳定性。方法采用高效液相色谱法测定克林霉素与酚磺乙胺在6 h内的含量变化,同时观察外观及测定pH值的变化。结果各配伍液在测定时间内无明显外观、pH值、峰形及保留时间的变化。克林霉素与酚磺乙胺在配伍液中的含量保持在初始浓度的97%以上。结论克林霉素与酚磺乙胺配伍后6 h内比较稳定,可以配伍。     

Analysis of major antioxidants from extracts of Myrmecodia pendans by UV/visible spectrophotometer,liquid chromatography/tandem mass spectrometry,and high-performance liquid chromatography/UV techniques

In the present work, heat reflux extraction with ethanol/water (80:20; v/v) as the solvent was used to extract antioxidants from Myrmecodia pendans. The crude extract (CE) was fractionated using hexane and ethyl acetate. Ethyl acetate fraction (EAF) and aqueous fraction were collected. Antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl-radical radical and ferric reducing power of the CE, EAF, and aqueous fraction were evaluated. EAF showed comparable antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl-radical radical and ferric reducing power to those of the CE. UV/visible, liquid chromatography/electrospray ionization/tandem mass spectrometry, and high-performance liquid chromatography were employed for identifying the major antioxidant compounds in the EAF. Three major phenolic compounds (rosmarinic acid, procyanidin B1, and polymer of procyanidin B1) were identified. The first two compounds were confirmed and quantified by high-performance liquid chromatography using authentic standards, but confirmation of the third compound was hampered by a lack of commercial standard. Concentrations of rosmarinic acid and procyanidin B1 in the EAF were found to be 20.688 ±1.573 mg/g dry sample and 3.236 ±0.280 mg/g dry sample, respectively. All these three compounds are reported for the first time in sarang semut.     

8名健康志愿者静滴头孢西丁的药物动力学

目的:测定国产头孢西丁iv gtt在人体内的药物动力学.方法:8例健康志愿者,恒速iv gtt 2g国产头孢西丁,用高效液相色谱外标法测定国产头孢西丁在正常人体内的血药浓度.结果:头孢西丁血浓峰值为(92.07±21.95)μg/ml,T_(1/2β)为(54.29±8.01)min,Cl为(0.38±0.08)L/min,Vd为(13.27±3.69)L.AUC为(90.34±19.59)μg/(h·ml),K_(10)为(0.029±0.0036)L/min.各项动力学参数与国外文献报道基本相符.结论:头孢西丁国产与进口产品的体内处置过程相同.     

Oestrogenic potency of genistin and daidzin in mice

The oestrogenic activity of genistin and daidzin was investigated in the B6D2F1 strain of mouse. Diethylstilboestrol and genistein were used as positive controls. The oestrogenic response to 1.5 mg genistin was equivalent to that of 1 mg genistein, giving a 1:1 molar relationship in oestrogenic activity between genistin and genistein. The oestrogenic response to 3.8 mg daidzin was equivalent to that of 1 mg genistein. Analysis of the blood obtained from the mice in this experiment failed to reveal any free genistein or daidzein in the plasma.     

Column-Switching High-Performance Liquid Chromatographic (HPLC) Determination of Hydrochlorothiazide in Rat,Dog, and Human Plasma

A fully automated HPLC assay for hydrochlorothiazide in plasma has been developed using a column-switching technique. The method involves direct injection of plasma to the extraction column for sample cleanup followed by switching onto the analytical column. Good precision, accuracy, and linearity were obtained over a range of 25 to 2000 ng/ml in rat, dog, and human plasma. The column-switching method has also been validated by comparison with a conventional HPLC method requiring a cumbersome plasma extraction procedure. Since the method is simple, rapid, and reproducible, it is useful for determination of hydrochlorothiazide levels in animal and human plasma.     

西沙必利片剂和胶囊剂人体药代动力学及相对生物利用度研究

目的研究西沙必利片剂及胶囊剂在健康人体内的药代动力学及相对生物利用度。方法采用反相高效液相色谱法测定12名健康自愿者单剂量口服三种制剂各15mg后,血浆中西沙必利浓度变化情况,用3P97程序按非室模型进行药代动力学分析。结果测得主要药代动力学参数为试验片剂T