Amelioration of Disordered Hepatic Protein Synthesis by the Deleted Form of Hepatocyte Growth Factor in Models of Liver Failure in Rats

Because the liver plays an important role in protein synthesis and cholesterol metabolism and reductions in these functions are observed in almost all hepatic disorders, the effects of the deleted form of hepatocyte growth factor (dHGF) on disordered hepatic protein synthesis were studied in various liver-injured rat models using Wistar male rats. In the 70% hepatectomized rats, plasma clotting time was prolonged and the serum level of total protein and the liver protein content were decreased. The treatment of the animals with dHGF (100?500 μg kg^?1, i.v., twice daily) ameliorated these parameters at 48 or 72 h. The administration of carbon tetrachloride or D-galactosamine to hepatectomized rats induced a marked prolongation of plasma clotting time and hypoproteinaemia. In the animals treated with dHGF (500 μg kg^?1, i.v., twice daily) these parameters were rapidly reversed compared with those of control groups. In a hepatocellular necrosis model induced by dimethylnitrosamine, the plasma clotting time was extremely prolonged, and liver protein content, serum total protein, albumin, HDL-cholesterol (as an index of lipoprotein) and plasma lecithin-cholesterol acyltransferase activity severely reduced. In this severely injured model, dHGF (5?500 μg kg^?1, i.v., twice daily for 28 days) dose-dependently prevented the loss of liver protein content and improved the disordered plasma coagulability and serum protein levels. These results suggest that dHGF is useful for ameliorating the disorders in hepatic functions such as protein synthesis.     

肝细胞生长因子在急性大鼠肺栓塞中的表达

目的:研究大鼠急性肺栓塞时肝细胞生长因子(HGF)的表达情况,评价HGF在急性肺栓塞中的诊断价值,并探讨其机制.方法:采用自体血栓回输法制作急性大鼠肺栓塞动物模型,分别设血浆实验组、组织实验组及其相应对照组,观察急性肺栓塞不同时间点血浆HGF水平和造模后12 h组织HGF mRNA的表达.结果:血浆实验组在不同时间点1、7、12和24 h HGF(μg/L)的表达(1.47±0.32、2.16±0.06、2.84±1.25和2.03±0.80)明显高于对照组的0.35± 0.07、0.38±0.09和0.37±0.07,差异有统计学意义(t分别为9.687 6,8.328 9,5.546 5和5.875 3,P ＜ 0.01),组织实验组在肺栓塞12 h时为(3.42±0.56)×10~(-2),与对照组(1.32±0.34)×10~(-2)相比,HGF mRNA表达差异有统计学意义(t ＝ 3.731 7, P ＜ 0.01).结论:HGF在急性肺栓塞时表达升高,可以作为诊断急性肺栓塞的一项指标.     

注射用促肝细胞生长素的细菌内毒素检查法研究

目的 :确定注射用促肝细胞生长素 (HGF)可用细菌内毒素检查法代替传统的热原检查法。方法 :用鲎试剂和家兔热原法做对比。结果 :将注射用HGF稀释成 0 .2 5mg/ml,用灵敏度为 0 .12 5EU/ml的鲎试剂进行内毒素检查 ,能有效消除干扰。结论 :细菌内毒素检查法可以代替热原检查法 ,对HGF进行质量控制。     

Effect of Route of Administration of Human Recombinant Factor VIII on Its Immunogenicity in Hemophilia A Mice

Factor VIII is a multi-domain glycoprotein and is an essential cofactor in the blood coagulation cascade. Its deficiency or dysfunction causes Hemophilia A, a bleeding disorder. Replacement using exogenous recombinant Factor VIII (FVIII) is the first line of therapy for Hemophilia A. Immunogenicity, the development of binding (total) and neutralizing (inhibitory) antibody against administered protein is a clinical complication of the therapy. There are several product related factors such as presence of aggregates, route and frequency of administration and glycosylation have been shown to contribute to immunogenicity. The effect of route of administration of FVIII on antibody development in Hemophilia A is not completely understood. Here we investigated the effect of route of administration (s.c. or i.v.) on immunogenicity in Hemophilia A mice. The total and inhibitory titers were determined using ELISA and modified Bethesda Assay respectively. The results indicated that s.c. is more immunogenic compared to i.v. route in terms of total antibody titer development (binding antibodies) but no significant differences in inhibitory titer levels could be established. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4480–4484, 2009     

急性肝衰竭大鼠血浆内毒素水平的检测及其与肝细胞生长素的关系

本实验采用鲎试验基质显色法检测D-氨基半乳糖(GaLN)形成急性肝衰竭(FHF)模型大鼠血浆内毒素(LPS)水平,并探讨了肝细胞生长素(HGF)对其影响。结果表明:FHF早期血浆LPS水平较正常对照组水平(49±6.94pg/ml)明显升高,且其水平随时间延长而自然降低。动态观察第9小时为高峰期(234±83.8pg/ml),30小时为最低水平(63.1±18.4pg/ml),与正常对照组比较分别具非常显著(P<0.01)或显著性差异(P<0.05)。结合肝组织病理学检查结果提示,内毒素血症出现在形成明显的肝损害之前。以HGF早期一次性腹腔注射治疗,发现9小时治疗组血浆LPS水平(149±41.9pg/ml)明显低于盐水对照组水平(234±83.8pg/ml,P<0.005)。相反,24小时HGF组LPS水平升高(266±55.9pg/ml)而明显高于盐水对照组水平(153±71.9pg/ml,P<0.001)。改变给药方式,则24小时两组LPS水平比较无显著性差异(115±14.5pg/ml与117±12.1pg/ml,P>0.05)。本研究进一步说明HGF、内毒素血症及肝损伤三者间紧密联系,并提示HGF治疗FHF所出现的内毒素血症以间歇或持续用药为宜。     

肝细胞生长因子对周围神经再生的作用研究

目的：研究肝细胞生长因子（HGF）对周围神经再生的影响。方法：查阅有关献，综述HGF的结构以及在周围神经系统发育、损伤修复中的作用，并提出临床应用中的问题。结果：HGF在周围神经系统再生方面有一定的作用，但其作用机制、给药方式及途径有待进一步研究。结论：随着医学研究的进步，会增加HGF用于周围神经再生的临床治疗可能性。     

Pharmacokinetic Modeling of Hepatocyte Growth Factor in Experimental Animals and Humans

Hepatocyte growth factor (HGF) is under development for treatment of renal failure. This study was designed to clarify changes in HGF pharmacokinetics in renal failure and to establish a pharmacokinetic model applicable to single and repeated doses. The plasma concentration profile in mice with glycerol‐induced acute renal failure was similar to that in normal mice, indicating a minimal contribution of kidney to systemic clearance of HGF. Nevertheless, accumulation of fluorescein‐4‐isocyanate‐labeled HGF in renal tubules in both cases suggests the occurrence of efficient endocytosis of HGF in kidney. A pharmacokinetic model including plasma and liver compartments was constructed, incorporating both high‐ and low‐affinity receptors for association and subsequent endocytosis of HGF because HGF is eliminated via specific receptor c‐Met and heparin‐like substance. The model well explained the plasma concentration profiles at all doses examined after bolus injection in animals and humans, and those during infusion in rodents. It includes externalization of receptors, which is negatively regulated by HGF, and can explain the gradual increase in trough concentration during repeated dosing in monkeys. Overall pharmacokinetic profiles of HGF are governed by at least two receptors and are well described by this pharmacokinetic model, which should assist in safe management of clinical trials.     

携带人肝细胞生长因子基因重组腺病毒的构建与制备

构建一种携带人肝细胞生长因子基因的重组腺病毒(AdHGF),并对其进行扩增、纯化与质量检测。首先构建携带人肝细胞生长因子基因的穿梭质粒pXCJL1CMV/pAHGF,然后用LipofectAMIN介导该质粒和含有E1、E3区及包装信号区缺失的复制缺陷型5型腺病毒基因组的质粒GT4050共转染293细胞,采用细胞内质粒DNA同源重组法构建重组腺病毒(AdHGF)。并以噬斑分析法筛选单克隆重组腺病毒,PCR法鉴定阳性重组腺病毒,氯化铯密度梯度离心法纯化病毒,紫外分光光度仪测定病毒颗粒数及纯度,快速CPE分析、噬斑分析法测定病毒感染滴度(pfu/ml)。采用敏感细胞病变法检测有复制能力的腺病毒。结果成功构建了重组腺病毒AdHGF,制备的病毒纯度好、滴度高,其效价比小于1∶100,并且未检测到有复制能力的腺病毒存在。研究构建制备的重组腺病毒AdHGF具有潜在的实际应用价值。     

Controlled Release of Hepatocyte Growth Factor from Gelatin Hydrogels Based on Hydrogel Degradation

This paper investigates the controlled release of hepatocyte growth factor (HGF) by biodegradable gelatin hydrogels and their HGF-induced angiogenic effect. Hydrogels of different degradabilities were prepared through chemical crosslinking gelatin with varied amounts of glutaraldehyde. When the gelatin hydrogels were radioiodinated and subcutaneously implanted into the back of mice, the remaining radioactivity of the hydrogels decreased with time. However, the remaining period became longer when the concentration of glutaraldehyde used for hydrogel preparation increased. Following implantation of gelatin hydrogels incorporating ¹²⁵I-labeled HGF, the HGF radioactivity retained in the mouse subcutis for longer time periods as the glutaraldehyde concentration becomes higher. The time profile of HGF remaining in every gelatin hydrogel was in good accordance with that of hydrogel degradation, indicating HGF release as a result of hydrogel biodegradation. The gelatin hydrogel incorporating HGF histologically induced angiogenic change around the implanted hydrogel. Gelatin hydrogels incorporating 5 and 10 jig HGF significantly enhanced the number of capillaries newly formed around the implanted site. This was in marked contrast to free HGF of same dose form and HGF-free, empty gelatin hydrogel. The gelatin hydrogel incorporating HGF induced VEGF around the implanted site. In vitro bioassay revealed that HGF molecules interacting with gelatin, still exhibited the biological activity. The interacted HGF would be released from gelatin hydrogels only when they were degraded to generate water-soluble gelatin fragments. It is possible that the HGF associating gelatin fragments of bioactivating, results in induced angiogenic effect.     

注射用促肝细胞生长素辅料的研究

寻找一种注射用促肝细胞生长素（ＨＧＦ）适用的辅料。方法：以三批不同批次的ＨＧＦ原料液为实验对象,分别通过 Ｆｏｌｉｎ－酚和~３Ｈ－ＴｄＲ掺入法研究了三种辅料： ３％右旋糖酐, ８％甘露醇, ２％右旋糖酐＋ ８％甘露醇对ＨＧＦ多肽含量及活性的影响。结果：８％甘露醇作为辅料时使ＨＧＦ多肽含量测定显著增高,而右旋糖酐则不影响测定结果。三种辅料对ＨＧＦ活性均无显著影响。结论：最终选用３％右旋糖酐作为ＨＧＦ的辅料。     

不同剂型重组人表皮生长因子对烧伤创面促愈合作用比较

目的比较重组人表皮生长因子(rhEGF)凝胶剂和水溶液剂治疗烧伤创面的临床疗效.方法选择烧伤浅Ⅱ°、深Ⅱ°和新鲜供皮区创面为受试对象,将创面分成A、B两区,采用随机自身对照试验.试验组用重组人表皮生长因子凝胶治疗,对照组用重组人表皮生长因子水溶液治疗,每日换药一次,观察不同时间内创面动态愈合率及愈合时间.结果重组人表皮生长因子凝胶治疗的烧伤浅Ⅱ°、深Ⅱ°和新鲜供皮区创面其愈合时间较重组人表皮生长因子水溶液组有明显缩短(P<0.01);动态愈合率也明显加快,组间具有非常显著性差异.结论重组人表皮生长因子凝胶剂用于烧烫伤、新鲜供皮区创面的冶疗均有较好的促进修复作用,临床效果优于重组人表皮生长因子水溶液剂组,提示重组人表皮生长因子的剂型是影响临床效果的重要因素,重组人表皮生长因子的药理作用因剂型的不同而存在差异.     

还原型谷胱甘肽联合促肝细胞生长素治疗慢性重型肝炎的疗效研究

目的 探讨还原型谷胱甘肽联合促肝细胞生长素治疗慢性重型肝炎的疗效.方法 选择简阳市人民医院2010年1月-2012年1月间收治的100例慢性重型肝炎患者为研究对象,随机将其分为对照组和观察组,两组患者均接受综合性治疗,观察组在此基础上接受还原型谷胱甘肽联合促肝细胞生长素治疗,对比分析两组患者临床疗效.结果 观察组患者临床治疗后PT、TBiL、AST和ALT生化指标均低于对照组,观察组患者临床治疗后的总有效率（74%）高于对照组（54%）,且差异均有统计学意义（P＜0.05）.结论 慢性重型肝炎患者在常规临床治疗措施的基础上配伍还原型谷胱甘肽联合促肝细胞生长素治疗,有助于患者临床治疗效果的改善,值得临床应用与推广.     

Characterization of the Enhancing Effect of Protamine on the Proliferative Activity of Hepatocyte Growth Factor in Rat Hepatocytes

Purpose  The aim of the present study was to characterize the mechanism of the stimulatory effect of protamine on HGF activity. Methods  The enhancing effects of protamine on the proliferative activity of HGF were investigated in vivo, in primary cultured rat hepatocytes, and in perfused rat liver. Results  In α-naphthylisothiocyanate-intoxicated rats, pretreatment with protamine increased HGF-induced autophosphorylation of the HGF receptor in liver. The maximum enhancing effect of protamine on HGF-induced DNA synthesis of hepatocytes required a 10 min-pretreatment period both in vivo and in vitro, and the stimulatory effect of protamine was not observed when it was administered simultaneously with HGF. Preperfusion of the liver with protamine for 10 min decreased the non-saturable portion of hepatic clearance for ¹²⁵I-HGF, which is mainly mediated by cell-surface heparan-sulfate proteoglycan (HSPG). Inhibition of HGF binding to heparin by protamine was confirmed using heparin-coated sepharose. This inhibition also required 10 min of pretreatment, for protamine to bind heparin. Conclusion  The enhancing effect of protamine on the mitogenic activity of HGF on hepatocytes requires pretreatment with protamine for a short period presumably required for its binding to cell-surface heparin, implying possible regulation of c-met autophosphorylation by HSPG.     

Enhancement of Memories by Systemic Administration of Insulin-Like Growth Factor II

To treat cognitive disorders in humans, new effective therapies that can be easily delivered systemically are needed. Previous studies showed that a bilateral injection of insulin-like growth factor II (IGF-II) into the dorsal hippocampus of rats or mice enhances fear memories and facilitates fear extinction. Here, we report that, in mice, systemic treatments with IGF-II given before training significantly enhance the retention and persistence of several types of working, short-term and long-term memories, including fear conditioning, object recognition, object placement, social recognition, and spatial reference memory. IGF-II-mediated memory enhancement does not alter memory flexibility or the ability for new learning and also occurs when IGF-II treatment is given in concert with memory retrieval. Thus IGF-II may represent a potentially important and effective treatment for enhancing human cognitive and executive functions.     

微波灼伤创面应用生长因子后的DNA含量分析

目的:探讨重组人酸性成纤维细胞生长因子对微波灼伤创面组织细胞的促增殖作用。方法:运用NS-FⅡ型多功能光谱治疗仪器建立微波灼伤动物模型,分为5组,每个创面按分组分别滴加0．9％氯化钠注射液(NS)、重组人表皮生长因子(rh-EGF,5 mg·L-1)、重组人酸性成纤维细胞生长因子(rh-aFGF,5mg·L-1)、重组人碱性成纤维细胞生长因子(rh-bFGF,5 mg·L-1)和牛脑提取液(bBE,1O mg·L-1)各0．2 ml,直到伤后第5d,取创面组织经流式细胞仪检测全层皮肤组织细胞周期及DNA含量。结果:应用生长因子各组的SPF、PI及DNA含量均明显高于NS组(P<0．05)。结论:重组人酸性成纤维细胞生长因子具有促进微波灼伤创面组织细胞增殖作用,促进创面的修复,缩短创面愈合时间。     

浅论非洛地平的药理作用及临床应用

目的观察钙离子拮抗剂非洛地平的药理作用及治疗肾性高血压临床效果。方法选取148例肾性高血压患者均采用口服非洛地平片,2.5mg/次,2次/d,疗程为2个月。治疗后观察患者的血压、血、尿蛋白(β2-MG)、肾功能(BUN、Scr、UA、Ccr、24h尿蛋白定量)、不良反应及服药情况等。结果 148例患者在经过非洛地平治疗后收缩压、舒张压、血肌酐、尿素氮、肌酐清除率、血尿酸、24h尿蛋白定量比较差异均有统计学意义(P<0.05)。结论非洛地平对治疗肾性高血压有良好的效果。     

Contribution of Parenchymal and Non-Parenchymal Liver Cells to the Clearance of Hepatocyte Growth Factor From the Circulation in Rats

Purpose. The distribution of ¹²⁵I-hepatocyte growth factor (HGF) to either liver parenchymal cells (PC) or non-parenchymal cells (NPC) was investigated in rats. Methods. After injection of a trace amount of ¹²⁵I-HGF, the distribution of radioactivity determined by microautoradiography closely resembled that of ¹²⁵I-epidermal growth factor which distributes mainly to PC. Results. The uptake clearance of ¹²⁵I-HGF estimated by determining the radioactivity of isolated liver cells was three times higher for PC than for NPC. This suggests that HGF distributes mainly to PC at relatively low doses. On the other hand, the uptake clearance by PC fell on coadministering an excess (80 µg/kg) of unlabeled HGF, while no change was observed for NPC, indicating that a saturable process for the hepatic handling of HGF exists only in PC where the HGF receptor is expressed. Conclusions. At such a dose the uptake clearance was comparable for both PC and NPC showing that HGF distributes to both cell types although NPC have few HGF receptors. Since the distribution to NPC was relatively non-specific and heparin-sensitive, it may be that heparin-like substances, which are believed to exist on PC and/ or the extracellular matrix, also exist on NPC.     

口服促肝细胞生长素微球的研究

以邻苯二甲酸醋酸纤维素为成球材料、促肝细胞生长素为球心物，制备工艺经均匀设计优化，制得类白色粉末状微球。高效液相色谱（HPLC）测得微球包封的药物图谱与促肝细胞生长素的HPLC图谱一致。微球载药量量29．7±2．5％，平均粒径127um。微球在人工胃液中2h释药不超过10％，在人工肠液中2h，释药100％。微球给大鼠灌胃后，HPLC测得大鼠血中促肝细胞生长素浓度在2．5h最高。     

鱼腥草的有效成分、药理作用及临床应用的研究进展

鱼腥草中含有癸酰乙醛、癸醛、月桂醛及甲壬酮等成分，其中癸酰乙醛具有抗病原微生物活性．药理研究证明鱼腥草具有抗菌、抗病毒、增强机体免疫、抗炎等作用，故在呼吸科、五官科及消化科疾病中都有广泛的应用．