当归多糖抑制慢性炎症贫血大鼠体内铁调素的表达

目的:探讨不同浓度当归多糖对慢性炎症贫血(ACD)大鼠模型铁调素(hepcidin)表达的影响。方法:通过对SD雄性大鼠足跖注射100 μL结核分枝杆菌的弗氏完全佐剂(CFA),建立ACD动物模型。利用此模型比较药物干预组(当归多糖)与非干预组及正常对照组大鼠肝脏内hepcidin表达的差异。结果:ACD组大鼠hepcidin的表达显著增高,贫血明显;灌胃当归多糖(ASP)后,hepcidin的表达降低,贫血改善。结论:当归多糖可能通过促红细胞生成素(EPO)相似途径抑制炎症反应,减少炎症细胞因子的产生。同时抑制hepcidin的表达提高ACD模型的铁含量,有利于阻止炎症因子与贫血的发展,为ACD的治疗提供一条新策略。     

当归多糖对H22荷瘤小鼠肿瘤生长及体内铁代谢的影响

目的:研究当归多糖(Angelica sinensis polysaccharide,ASP)对H22荷瘤小鼠肿瘤生长及体内hepcidin和IL-6表达的抑制作用。方法:移植H22鼠肝癌细胞于小鼠右腋皮下,建立H22荷瘤小鼠模型,分为空白组(不接种不给药),阴性对照组(腹腔注射等体积生理盐水),阳性对照组(腹腔注射环磷酰胺,25 mg·kg^-1),ASP高剂量组(灌胃ASP溶液,100 mg·kg^-1),ASP中剂量组(灌胃ASP溶液,50 mg·kg^-1),ASP低剂量组(灌胃ASP溶液,25 mg·kg^-1),连续给药14 d,末次给药24 h后摘眼球取血,剥离肿瘤、脾脏并称重,采用ELISA法检测血清中hepcidin和IL-6的含量变化。结果:3个剂量组当归多糖均有一定的抗肿瘤效果,抑瘤率分别为24.12%,29.15%和34.67%,同时能显著抑制hepcidin和IL-6的表达。结论:当归多糖的体内抗肿瘤作用可能与其对铁代谢的调节有关。     

促红细胞生成素抑制正常及缺铁性贫血大鼠铁调素的表达

目的:探讨促红细胞生成素(EPO)在缺铁性贫血(IDA)大鼠模型中对铁调素(hepcidin)表达的影响。方法:通过反复放血和饲喂低铁饲料,建立IDA大鼠动物模型,比较药物干预组(EPO)与非干预组及正常对照组与正常给药组(EPO)大鼠血清hepcidin表达的差异。Western blot检测肝脏内信号因子CCAAT/增强子结合蛋白α(C/EBPα)含量。结果:IDA模型组腹腔注射EPO后,血清hepcidin和肝脏C/EBPα含量显著降低。EPO对正常大鼠hepcidin表达也有显著抑制作用(P<0.05)。结论:hepcidin可能参与了IDA的发生,在正常大鼠和IDA大鼠中,EPO能通过下调C/EBPα阻碍其激活hepcidin启动子最终抑制hepcidin的表达。     

五味子多糖对原代大鼠肝细胞氧化应激损伤的保护机制研究

目的探讨五味子多糖对原代大鼠肝细胞氧化应激的保护效应,并对其作用机制进行研究。方法体外培养原代大鼠肝细胞,分为空白对照组、H2O2诱导组、H2O2+低剂量五味子多糖组、H2O2+中剂量五味子多糖组、H2O2+高剂量五味子多糖组;CCK-8试剂检测大鼠肝细胞相对存活率,相关试剂盒检测NO、MDA含量,Western blot检测大鼠肝细胞中P67-phox、P47-phox、SOD1、HO-1、Rac1、p-Rac1蛋白表达水平。结果与H2O2诱导组相比,不同浓度五味子多糖可明显提升大鼠肝细胞的存活率(P<0.05);与H2O2诱导组相比,不同剂量五味子多糖可明显抑制细胞中NO、MDA的生成(P<0.05);Western blot检测显示,不同剂量五味子多糖可明显促进抗氧化蛋白HO-1、SOD1表达,抑制促氧化蛋白P67-phox、P47-phox的表达,并且五味子多糖显著抑制Rac1的磷酸化(P<0.05)。结论五味子多糖可明显改善大鼠肝细胞氧化应激损伤,其作用机制可能与抑制Rac1的磷酸化有关。     

银杏叶提取物对重症胰腺炎胰腺细胞凋亡作用机制的探讨

目的　探讨银杏叶提取物 (EGb)对急性重症胰腺炎 (ASP)胰腺细胞凋亡的作用机制。方法　将 78只SD大鼠分为假手术 (SO)组、ASP组和治疗组。治疗组在ASP模型诱发后经腹腔注射金纳多注射液 2 0mg/kg ,每 6h注射一次 ,动态观察胰腺病理变化 ,免疫组化 (TUNEL法与SP法 )检测胰腺凋亡情况与细胞凋亡调控基因Bcl 2和C myc蛋白的表达。结果　治疗组胰腺病理损害程度较ASP组减轻。ASP组凋亡指数明显高于SO组 ,(P <0 0 1) ,且术后 3h达高峰 ,治疗组 3h、6h凋亡指数明显下降 (P <0 0 5 )。免疫组化 :ASP组Bcl 2表达明显增多 ,治疗组 3h的Bcl 2表达较ASP组同时相点上调 (P <0 0 5 ) ;ASP组C myc表达术后 6h达高峰 ,治疗组 3h、12h的C myc表达虽有增高 ,但无显著性差异。结论　金纳多能减轻胰腺病理损害、抑制胰腺细胞的凋亡 ,其抑制细胞凋亡可能与其上调凋亡调控基因Bcl 2的表达有关。     

柱前衍生化HPLC-FD法测定大鼠体内当归多糖组分ASP1的含量

摘 要 目的：建立高效液相色谱 荧光检测法（HPLC-FD）测定大鼠体内当归多糖ASP1含量的方法,为其药代动力学研究奠定基础。方法: 采用Belder 和Granath的方法将异硫氰酸荧光素（FITC）标记到精制当归多糖组分ASP1上,得到标记产物ASP1-FITC;组织样品经30%三氯乙酸溶液和11%氢氧化钠溶液处理后进样。采用HPLC-FD法,以PL aquagel-OH MIXED（300 mm×7.5mm,8 μm）为色谱柱,以pH 7.0的磷酸盐缓冲液（取磷酸二氢钠2.34 g、磷酸氢二钠4.33 g与氯化钠11.70 g,加水使溶解成1000 ml）为流动相,流速0.5 ml·min^-1,荧光检测器λex 494 nm、λem 520 nm。结果: ASP1-FITC在组织样品中的定量下限为0.20 μg·ml^-1,在0.25～40.00 μg·ml^-1范围浓度内与峰面积呈良好的线性关系（r﹥0.999 6）,回收率在91.98%～114.20%之间,日内精密度RSD和日间精密度RSD分别低于8.31%和2.94%,均符合要求。结论: 本试验建立了当归多糖ASP1柱前荧光标记衍生、HPLC-FD法测定大鼠组织样品中当归多糖组分ASP1含量的方法,该方法样品用量少、灵敏度高、特异性强、精密度和重复性较好,适用于当归多糖组分ASP1药代动力学和组织分布试验的检测。     

硝普钠抑制脂多糖诱导的大鼠原代培养肝细胞表面细胞间粘附分子-1的表达

目的　评价模拟肝脏急性炎症期一氧化氮与细胞间粘附分子 1(ICAM 1)表达的关系。方法　用免疫细胞化学技术检测加入不同浓度NO供体硝普钠 (SNP)和 (或 )细菌内毒素 (LPS)后 ,原代培养大鼠肝细胞ICAM 1的表达情况。结果　当培养时间达4h ,各组肝细胞均未见ICAM 1显著表达 ;但当培养至 8,2 4h ,LPS组的肝细胞ICAM 1表达强度明显升高 (P <0 .0 5 ) ,SNP则可呈浓度依赖性地抑制LPS诱导的大鼠肝细胞ICAM 1的表达。结论　SNP可显著抑制LPS诱导的原代培养大鼠肝细胞ICAM 1表达。     

大鼠原代培养肝细胞免疫性损伤的病理特征及甘利欣的护肝作用

目的:探讨大鼠原代培养肝细胞免疫性损伤模型病理特征,评价甘利欣(Grz)护肝作用。方法:用卡介苗(BCG)整体致敏,脂多糖(LPS)离体攻击(BCG+LPS)致肝细胞损伤。常规测AST,LDH活性,Griess法测NO含量;免疫细胞化学法测ICAM-1表达,丫啶橙荧光染色法和流式细胞仪测肝细胞凋亡率,评价Grz护肝作用。结果:LPS对上清AST无显著影响。BCG+LPS使上清AST,LDH,NO,肝细胞ICA8-1表达及凋亡率均明显高于正常对照组;Grz抑制AST和LDH的同时,使NO和12 h肝细胞凋亡率显著降低。氨基胍对NO也有显著抑制。结论:ICAM-1高表达,大量NO生成和肝细胞凋亡可能是BCG+LPS诱发大鼠原代肝细胞免疫性损伤的病理现象;Grz可显著抑制NO生成和肝细胞凋亡。     

当归多糖铁复合物在大鼠体内2种不同吸收途径的研究

目的:通过当归多糖铁复合物(APIC)2种不同方式给药来探讨其在大鼠体内以游离铁离子形式吸收与直接以多糖铁复合物分子形式吸收这2种不同吸收途径。方法:分别进行正常大鼠APIC高、中、低剂量组灌胃给药和十二指肠给药实验。以原子吸收法测定各时间点血清铁浓度,用DAS2.0软件求算药动学参数。结果:APIC在大鼠体内的药动学过程均符合二室模型,且在本实验所采用剂量范围AUC随着剂量的增加也相应增大。同等剂量灌胃给药,不加抗坏血酸组AUC比加抗坏血酸组AUC稍大。且APIC能以多糖铁复合物分子形式被十二指肠吸收。结论:APIC既能以游离铁离子形式又能以多糖铁复合物分子形式被十二指肠吸收。     

当归多糖对大鼠Kupffer细胞免疫功能的激活作用当归多糖对大鼠Kupffer细胞免疫功能的激活作用

目的观察当归多糖(ASP)对大鼠Kupffer细胞免疫功能的影响。方法ASP体外作用于正常Kupffer细胞,以吞噬中性红A₅₄₀、分泌NO和TNF-α量评价其免疫功能;以LDH漏出量,评价ASP对细胞的直接损害情况。大鼠ig ASP后,检测Kupffer细胞上述指标及胞内酸性磷酸酶(ACP)活性;以sALT和sGST为肝毒性指标。结果ASP增强Kupffer细胞对中性红吞噬作用、增加ACP活性及NO和TNF-α的产生。ASP在体外不增加肝实质细胞LDH漏出,而体内使sGST升高。结论适当剂量ASP可激活Kupffer细胞免疫功能。大剂量ASP出现的轻度肝功能改变可能与NO和TNF-α等免疫因子过度分泌间接有关。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Changes in angiopoietin expression in glomeruli involved in glomerulosclerosis in rats with daunorubicin-induced nephrosis

AIM: To study the potential pathological role of endogenous angiopoietins in daunorubicin-induced progressive glomerulosclerosis in rats. METHODS: Seventy male Wistar rats were allocated randomly into a daunorubicin group (DRB; n=40) or a control group (n=30). The rats in the DRB group were injected with DRB (15 mg/kg), in their tails. Subsequently, at intervals of 1, 2, 4, 6, 8, and 12 weeks, 5 male Wistar rats in each group were chosen randomly for 24 h urinary protein quantitative measurements (24 h UPQM), and determination of plasma tumor necrosis factor alpha (TNF-alpha), angiopoietin-1 (Ang1), and angiopoietin-2 (Ang2) levels. Kidney sections were examined by electron microscopy, Periodic Acid Schiff (PAS) staining, immunohistochemical staining and in situ hybridization histochemistry. RESULTS: As glomerulosclerosis progressed in the DRB group, expression of Ang1 mRNA and protein in glomeruli decreased and expression of TNF-alpha protein, Ang2 mRNA and protein in glomeruli increased. Expression of Ang1 mRNA and protein in glomeruli were negatively correlated with 24 h UPQM, Fn protein expression, and mean area of extracellular matrix (MAECM). In comparison, expression of Ang2 mRNA and protein in glomeruli were positively correlated with 24 h UPQM, Fn protein expression and MAECM; furthermore, there was a positive correlation between plasma Ang2 and 24 h UPQM. Plasma TNF-alpha and expression of TNF-alpha in glomeruli were positively correlated with expression of Ang2 mRNA and protein in glomeruli. There was a negative correlation between Ang1 protein expression and Ang2 protein expression in glomeruli. CONCLUSION: During DRB-induced glomerulosclerosis, podocyte injury led to a shift in the balance of Ang1 and Ang2 in glomeruli. Increased TNF-alpha in plasma and glomeruli may upregulate Ang2 expression in glomeruli. Elevated Ang2 in both plasma and glomeruli may mediate protein permeability through the glomerular filtration barrier. Moreover, local expression of Ang2 may facilitate the progress of glomerulosclerosis by upregulating a component expression of extracellular matrix.     

Deoxynivalenol in cereals in Russia

A survey of the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in wheat, rye, barley and maize harvested in 1989-2001 in several regions of Russia has been conducted. A total of 5652 samples of cereals were analysed for DON and ZEN by using TLC and normal-phase HPLC with UV-detector. DON was detected in 69% of 2166 samples from Krasnodar region which is considered to be the major Fusarium endemic region of Russia. The contamination levels ranged from 0.1 till 8.6 ppm, MTEL was exceeded in 37% of these samples. The positive correlation between DON concentration and a percentage of Fusaria-damaged wheat kernels has been shown. DON occurrence and contamination levels were much lower that for wheat. Based on the results of monitoring and the data of average actual consumption of wheat products in Russia, the estimated daily intake of DON per 1 kg of body weight (EDI)was calculated. EDI varied from 0.07 ug in 1990-1991 till 1.40 ug in 1992. Although average EDI were lower than adopted tolerable daily intake (TDI, 3 ug/kg body weight) EDIs for the North-Caucasian region in some cases exceeded TDI.