肝细胞生长因子对慢性肝损伤时肝部分切除术后肝再生和肝功能的影响

目的研究肝细胞生长因子对慢性肝损伤时肝部分切除术后肝再生和肝功能的影响。方法对慢性肝损伤模型SD大鼠行30%肝部分切除术,术后10d,经腹腔注射肝细胞生长因子,同时建立正常对照组。观察不同剂量肝细胞生长因子对肝再生,肝功能和肝储备功能的影响。结果实验组与对照组相比,AST,ALT,总胆固醇、动脉血酮体比具有显著的统计学意义(P<0.01)。结论应用外源性肝细胞生长因子可明显改善肝功能和肝储备功能,促进肝细胞再生。     

Stability of faecal hepatocyte growth factor determination

In order to evaluate the accuracy and reproducibility of determination of hepatocyte growth factor (HGF) levels in faeces, the stability of HGF in samples processed in different ways was investigated. An ELISA method was used for determination of HGF concentrations. Faeces samples from healthy controls and patients with infectious diarrhoea were studied. It was found that faeces HGF concentration remained stable irrespective of whether samples were freeze‐thawed several times, kept for 6, 12 or 24?h at room temperature or refrigerated for 6, 12, 24 or 36?h; the levels of HGF did not change significantly when samples were freeze‐dried. Adding protease inhibitor to the faeces samples did not affect the HGF levels. There were no significant differences between HGF levels using phosphate buffered saline (PBS) (pH?7.4) or NaCL as buffer, but it was observed that levels of HGF were significantly lower in the samples that were diluted in distilled water. Although both HGF and albumin through various mechanisms may increase in faeces during infectious diarrhoea, there was no significant correlation between faeces HGF levels and albumin levels, which might indicate local production of HGF in the bowel in response to infection. It is concluded that determination of faeces HGF levels is feasible with a high degree of stability. Increased HGF levels in faeces might represent a local production of HGF during bowel injury and might be of use as a diagnostic and monitoring assay.     

联合应用骨髓间充质干细胞与促肝细胞生长素治疗大鼠急性肝衰竭

目的 评价骨髓间充质干细胞(MSCs)和促肝细胞生长素(pHGF)联合治疗大鼠急性肝衰竭(ALF)的可行性.方法 将ALF造模成功的37只SD大鼠分为4组:CC14组(A组,9只),CCl4/MSCs(B组,9只),CC14/pHGF组(C组,9只)和CC14/pHGF+ MSCs组(D组,10只).用携带hrGFP基因的慢病毒感染MSCs(hrGFP-MSCs),并以hrGFP示踪.对C、D组大鼠给予CCl4,同时腹腔注射pHGF;B组和D组造模12h后肝内注射hrGFP-MSCs悬液.对各组大鼠分别在造模后24 h、72 h、7天检测血清丙氨酸转氨酶(ALT)水平,造模后72 h检测血清IL-6、IL-10、TNF-α炎症因子水平,造模后24 h取肝组织行HE染色,造模72 h行PCNA免疫组化染色检测肝细胞增殖情况.造模后72 h后取注射部位肝组织行冰冻切片观察移植hrGFP-MSCs肝内分布情况,对非注射部位肝组织进行实时荧光定量RT-PCR检测hrGFP基因含量.结果 联合治疗后,ALF大鼠外周血ALT水平降低,IL 6、IL-10、TNF-α炎症因子水平下调,肝脏病理组织学损害减轻,肝细胞增殖率增高.造模后72 h,B组与D组注射部位肝组织内可见移植的hrGFP-MSCs,非注射部位肝组织未见hrGFP荧光阳性细胞,但用实时荧光定量RT-PCR技术可在非注射部位肝组织内检测到hrGFP基因,且D组含量高于B组.结论 pHGF与MSCs联合治疗大鼠ALF的疗效优于MSCs或pHGF单独治疗,可能与pHGF促使其肝内定植MSCs增多有关.     

Plasminogen activator inhibitor‐1 inhibits factor VIIa bound to tissue factor

Summary. Background and objective: A growing body of experimental evidence supports broad inhibitory and regulatory activity of plasminogen activator inhibitor 1 (PAI‐1). The present study was designed to investigate whether PAI‐1 inhibits factor (F) VIIa complexed with tissue factor (TF), a well‐known procoagulant risk factor. Methods and results: The ability of PAI‐1 to inhibit FVIIa‐TF activity was evaluated in both clotting and factor X (FX) activation assays. PAI‐1 and its complex with vitronectin inhibit: (i) clotting activity of FVIIa‐TF (PAI‐1_IC50, 817 and 125 nm , respectively); (ii) FVIIa‐TF‐mediated FX activation (PAI‐1_IC50, 260 and 50 nm , respectively); and (iii) FVIIa bound to TF expressed on the surface of stimulated endothelial cells (PAI‐1_IC50, 260 and 120 nm , respectively). The association rate constant (k_a) for PAI‐1 inhibition of FVIIa‐TF was determined using a chromogenic assay. K_a for PAI‐1 inhibition of FVIIa bound to relipidated TF is 3.3‐fold higher than that for FVIIa bound to soluble TF (k_a = 0.09 ± 0.01 and 0.027 ± 0.03 μm ^?1 min^?1, respectively). Vitronectin increases k_a for both soluble and relipidated TF by 3.5‐ and 30‐fold, respectively (to 0.094 ± 0.020 and 2.7 ± 0.2 μm ^?1 min^?1). However, only a 3.5‐ to 5.0‐fold increase in the acylated FVIIa was observed on SDS PAGE in the presence of vitronectin for both relipidated and soluble TF, indicating fast formation of PAI‐1/vitronectin/FVIIa/relipidated TF non‐covalent complex. Conclusions: Our results demonstrate potential anticoagulant activity of PAI‐1 in the presence of vitronectin, which could contribute to regulation of hemostasis under pathological conditions such as severe sepsis, acute lung injury and pleural injury, where PAI‐1 and TF are overexpressed.     

Expression of hepatocyte growth factor, keratinocyte growth factor and their receptors in experimental chronic pancreatitis

BACKGROUND: Hepatocyte (HGF) and Keratinocyte growth factors (KGF) are key factors of tissue organization and regeneration. These peptide growth factors and their receptors c-met and keratinocyte growth factor receptor (KGFR) are overexpressed in pancreatic cancer. AIM: Expression and localization of ligands and receptors were investigated during the development of experimental chronic pancreatitis. METHODS: Chronic pancreatitis was induced in rats by intravenous injection of dibutyltin dichloride. One to 60 days after treatment, the expression of growth factors and receptors was analysed by competitive polymerase chain reaction, Western blot analysis and immunohistochemistry. RESULTS: HGF mRNA expression increased (10-fold) until days 7-14 followed by a decrease to control level. Expression of c-met mRNA constantly increased (15-fold). KGF and KGFR mRNA expression were increased after 14-28 days (5-fold) and then returned to control levels. mRNA expression patterns correlated with changes in the protein expression, whereas protein levels of KGF remained unchanged. Ligands were localized in mesenchymal cells and their receptors on epithelial cells. CONCLUSIONS: The significant increase of HGF and c-met expression suggests an essential role of this growth factor in the morphological changes during the development of chronic pancreatitis. Changes in the expression of KGF and KGFR are less pronounced.     

蛋白酶体抑制剂对多发性骨髓瘤患者骨髓间充质干细胞迁移能力及其肝细胞生长因子表达的影响

本研究探讨蛋白酶体抑制剂硼替佐米对多发性骨髓瘤(MM)患者骨髓间充质干细胞(MSC)迁移能力及其肝细胞生长因子(HGF)基因表达水平的影响。应用Transwell模型观察MM患者骨髓MSC经2.5 nmol/L硼替佐米处理前后的体外迁移能力,实时定量PCR检测MSC的HGF mRNA的表达水平。结果表明,经2.5 nmol/L的硼替佐米作用48小时后,MSC的迁移能力明显低于对照组,并且HGF mRNA在MSC的表达与对照组相比也明显降低,两者结果均具有统计学意义(p<0.05)。结论:硼替佐米可抑制MM患者骨髓MSC的迁移,同时可下调其趋化相关因子HGF的表达。     

急性白血病患者肝细胞生长因子及血管内皮生长因子血清水平检测及临床意义

目的 检测急性白血病(AL)患者治疗前后外周血血清肝细胞生长因子(HGF)和血管内皮生长因子(VEGF)的水平,探讨血清HGF和VEGF与急性白血病的发生、发展及预后等方面的关系.方法 应用双抗体夹心酶联免疫吸附测定(ELSIA)法动态检测并分析患者疾病不同时期血清HGF和VEGF的水平.结果 急性非淋巴细胞白血病(ANLL)组血清HGF水平(1 196.86±282.31)ng/L明显高于正常对照组(332.55±65.16)ng/L(P<0.01);急性淋巴细胞白血病(ALL)组血清HGF水平(1 224.53±283.19)ng/L明显高于正常对照组水平(332.55±65.16)ng/L(P<0.01);ANLL患者CR组治疗前后血清HGF水平分别为(1 140.31±266.23)ng/L,(478.50±183.75)ng/L(P<0.01),NR组治疗前后血清HGF水平分别为(1 432.51±235.30)ng/L,(1 386.63±197.95)ng/L(P>0.05).结论 AL患者血清HGF及VEGF水平均较正常对照组HGF水平及VEGF水平升高.血清HGF及VEGF水平可作为监测AL的疗效指标.血清VEGF水平可作为AL的预后判断指标,血清HGF水平对白血病的预后判断有一定意义.     

两种细胞因子受体在胰腺癌中的表达及临床应用价值

目的探讨胰腺癌组织中表皮生长因子受体（EGFR）和肝细胞生长因子受体（Met）基因和蛋白的表达情况,及其与胰腺癌临床病理特征和预后的关系。方法采用免疫组化法检测78例胰腺癌组织和23例正常胰腺组织中EGFR和Met蛋白的表达情况,采用实时定量聚合酶链反应（RT-RCR）法检测 EGFR和Met-DNA的相对拷贝数。结果78例胰腺癌组织中,EGFR和Met蛋白的阳性表达率明显高于正常胰腺组织,差异均有统计学意义（ P＜0．05）。肿瘤直径大于4 cm患者、胰腺癌组织高中分化患者、出现远处转移患者、淋巴转移患者、肠系膜上血管侵犯患者、T N M分期为Ⅲ和Ⅳ期患者中EG FR和M et蛋白的表达均明显高于肿瘤直径小于或等于4 cm患者,胰腺癌组织低分化患者,未出现远处转移患者、未有淋巴转移患者、未出现肠系膜上血管侵犯患者、TNM 分期为Ⅰ和Ⅱ期患者,差异具有统计学意义（P＜0．05）。胰腺癌患者中,EGFR和Met-DNA 相对拷贝数均明显高于正常胰腺组织中EGFR和Met-DNA相对拷贝数,差异具有统计学意义（P＜0．05）。高表达EGFR和Met胰腺癌患者的平均生存时间均显著低于Met及EGFR低表达患者,差异具有统计学意义（P＜0．05）。Met和EGFR均高表达的患者的平均生存时间较Met或 EGFR单一高表达者的短,差异具有统计学意义（P＜0．05）。结论 EGFR和M et的表达与胰腺癌的临床病理相关指标密切相关,对二者的分析研究有助于对胰腺癌患者的预后作出预测,并对化疗药物新靶点筛选提供理论依据。     

慢性乙型肝炎患者血清转化生长因子β1、肝细胞生长因子的检测及临床意义

目的探讨慢性乙型肝炎病理过程中血清转化生长因子β1(TGF-β1)、肝细胞生长因子(HGF)水平的变化及其临床意义。方法48例慢性乙型肝炎患者和20例健康人均于清晨空腹采血以双抗体夹心酶联免疫吸附测定法检测血清TGF-β1、HGF水平并同时检测肝功能和乙型肝炎病毒脱氧核糖核酸(HBV DNA)水平。结果慢性乙型肝炎患者轻度组、中度组、重度组血清TGF-β1、HGF水平均显著高于对照组,TGF-β1分别为(167.5±87.6)μg/L、(313.1±96.3)μg/L(、495.9±134.4)μg/L vs(81.4±40.7)μg/L,MGT分别为(1530.4±912.2)ng/L(、2461.5±1624.7)ng/L、(3805.2±2104.8)ng/L vs(270.3±123.5)ng/L(均P<0.05),且与丙氨酸转氨酶(ALT)、总胆红素(TBil)水平呈正相关(r=0.625,r=0.568,r=0.579,r=0.612);HBV DNA阳性组和阴性组血清TGF-β1、HGF水平比较差异无统计学意义。结论细胞因子TGF-β1、HGF参与了慢性乙型肝炎的病理生理过程,其水平变化与肝脏损伤程度密切相关。     

多发性骨髓瘤患者骨损害与肝细胞生长因子血清浓度关系的探讨

目的　探讨多发性骨髓瘤(MM)患者骨损害程度与肝细胞生长因子(HGF)血清浓度的关系。方法　采 用双抗体夹心酶联免疫吸附法检测32例MM患者的HGF血清浓度,观察其与多发性骨髓瘤骨损害的关系。结果　 不同程度骨损害的MM患者,其血清HGF浓度的差异有统计学意义(P<0.01);MM骨损害越严重,其血清HGF 浓度增高越显著。结论　MM患者HGF血清浓度与骨损害程度有关,骨损害程度严重者,HGF血清浓度明显增高。     

糖尿病大鼠肾小管上皮细胞转化与肝细胞生长因子和Smad7蛋白的表达

目的探讨糖尿病肾病（DN）时肾小管上皮细胞转化以及肝细胞生长因子（HGF）、Smad7蛋白在其中可能发挥的作用。方法60只Wistar大鼠均行右肾切除术，伤口愈合后随机分为右肾切除对照组和糖尿病组。腹腔注射链脲佐菌素诱发大鼠糖尿病模型。采用免疫组化法检测角蛋白18（CK18）、α-平滑肌肌动蛋白（α-SMA）、HGF和Smad7蛋白的表达；流式细胞术检测α-SMA和HGF的表达；蛋白质免疫印迹法检测Smad7蛋白的表达。结果糖尿病组较对照组CKl8的表达降低，α-SMA的表达上升，HGF和Smad7蛋白的表达表现为先上升后下降。结论糖尿病时，肾小管上皮细胞可发生表型转化，转化为肌成纤维细胞。HGF和Smad7蛋白表达下降可能与该模型中肾小管上皮细胞转化有关。     

脑出血对肝脏的影响及其临床意义的探讨

目的探讨脑出血患者血清中肝细胞再生因子的变化及其意义。方法应用组织细胞培养法及同位素标记技术对60例脑出血患者和30例健康对照组血清中肝细胞再生因子水平进行测定。结果≥30ml的脑出血的患者血清中肝细胞再生因子高于<30ml的脑出血的患者(P<0.05),并显著高于正常对照组(P<0.01)。结论为研究脑出血患者对肝脏的影响及其防治提供参考。     

正畸力作用下大鼠炎性牙周组织中肝细胞生长因子的表达

背景：细胞因子在牙周组织改建机制中具有重要的免疫调节和直接介导作用。目前肝细胞生长因子在正畸力作用后对牙周组织改建的作用及机制尚未见报道。 目的：探讨肝细胞生长因子在炎性牙周条件下参与牙移动及牙周改建的机制。 方法：选用30只8周龄雄性SD大鼠,建立牙周炎模型,随机分为2组,炎性对照组5只,炎性加力组25只。炎性加力组在上颌第一磨牙近中施加50 g的力值,分别在受力1,3,5,7,14 d处死,每次处死5只。采用苏木精-伊红染色、免疫组化方法分析牙齿受力移动不同阶段牙周组织中肝细胞生长因子表达和分布变化。 结果与结论：苏木精-伊红染色结果显示,受力牙齿与未受力牙齿牙周组织中都存在一定程度的改建。免疫组化结果显示,炎性对照组中可观察到肝细胞生长因子在牙周组织中呈阳性表达,分布较均匀。炎症加力组大鼠牙周组织内肝细胞生长因子受力后表达增高,在第5天达到高峰,之后开始下降;其中成骨细胞、成纤维细胞及破骨细胞均为强表达。提示肝细胞生长因子参与了正畸牙周组织的改建过程,其表达随时间呈现一定规律,炎症刺激可导致牙周组织中肝细胞生长因子的表达增高。     

基于间质表皮转化因子/肝细胞生长因子信号通路靶向分子成像探针研究进展

间质表皮转化因子(c-Met)/肝细胞生长因子(HGF)信号通路在肿瘤发生、发展中起重要作用.分子成像可准确、无创地评价关键分子靶点在体表达水平和活化状态.基于c-M et的肿瘤靶向分子成像不仅可检测c-M et表达状态以筛选受益患者,还能监测治疗效果、评估预后,提高靶向治疗效果.本文就基于c-Met/HGF信号通路的...     

A phase I/II exploratory clinical trial for intracordal injection of recombinant hepatocyte growth factor for vocal fold scar and sulcus

Vocal fold scar and sulcus are intractable diseases with no effective established treatments. Hepatocyte growth factor (HGF) has preclinically proven to have potent antifibrotic and regenerative effects on vocal fold scar. The current Phase I/II clinical trial aims to examine the safety and effectiveness of intracordal injection of a recombinant human HGF drug for patients with vocal fold scar or sulcus. This is an open‐label, dose‐escalating, first‐in‐human clinical trial. Eighteen patients with bilateral vocal fold scar or sulcus were enrolled and divided into three groups: Step I received 1 μg of HGF per vocal fold; Step II received 3 μg of HGF; and Step III received 10 μg of HGF. Injections were administered once weekly for 4 weeks. The protocol treatment was performed starting with Step I and escalating to Step III. Patients were followed for 6 months post‐treatment. Local and systemic safety aspects were examined as primary endpoints, and therapeutic effects were assessed as secondary endpoints using voice handicap index‐10; maximum phonation time; vocal fold vibratory amplitude; grade, rough, breathy, asthenic, strained scale; and jitter. The results indicated no serious drug‐related adverse events in either the systemic or local examinations. In whole‐subject analysis, voice handicap index‐10, vocal fold vibratory amplitude, and grade, rough, breathy, asthenic, strained scale were significantly improved at 6 months, whereas maximum phonation time and jitter varied. There were no significant differences in phonatory data between the step groups. In conclusion, intracordal injection of a recombinant human HGF drug was safe, feasible, and potentially effective for human patients with vocal fold scar or sulcus.     

黄芪对内毒素性急性肺损伤大鼠肝细胞生长因子表达的影响

目的：探讨黄芪在急性肺损伤（ALI）时对肝细胞生长因子（HGF）表达的影响，评价黄芪在ALI中的治疗机制。方法：制备内毒素（LPS）性ALI模型。设正常对照组（生理盐水1ml／kg）、模型组（LPS5mg／kg）、黄芪治疗组（黄芪8mg／kg）。用逆转录一聚合酶链反应（RT—PCR）检测各组大鼠肺组织匀浆中HGFmRNA表达的变化；用免疫组化法检测各组大鼠肺组织中HGF阳性细胞数的变化。结果：RT—PCR检测结果显示，ALl时HGF mRNA表达明显增加（P〈0．05），黄芪能促使HGFmRNA表达进一步升高（P〈0．05）。免疫组化显示，ALI时HGF阳性细胞表达产物增加（P〈0．01），黄芪能促进HGF的表达（P〈0．05）。结论：黄芪通过促进HGF的表达来促进ALI的修复。     

乳腺癌新辅助化疗对血清肝细胞生长因子水平影响的实验研究

目的探讨乳腺癌新辅助化疗对血清肝细胞生长因子水平的影响及相关临床意义。方法应用酶链免疫吸附法（ELIA）检测100例可手术乳腺癌患者新辅助化疗前及术后的血清肝细胞因子水平,并评估其水平与乳腺癌新辅助化疗疗效的关系。结果血清肝细胞生长因子水平与乳腺癌的分期密切相关（P=0．018）;血清肝细胞生长因子水平在治疗前后的变化与新辅助化疗效果密切相关。结论血清肝细胞生长因子可能在乳腺癌的发生发展中起重要作用,可能是预测化疗疗效的独立指标。     

血浆纤溶酶原激活物抑制物1及血清转化生长因子β与2型糖尿病肾病的相关性研究

目的 研究2型糖尿病肾病(DN)及单纯2型糖尿病(T2DM)患者血浆纤溶酶原激活物抑制物1(PAI-1)及血清转化生长因子β(TGF-β)水平的变化.并进一步探讨在T2DM患者中血浆PAI-1和血清TGF-β的关系.方法 T2DM患者93例,其中T2DM无蛋白尿患者(DM组)37例;微量蛋白尿患者(DN 1组)27例,尿白蛋白/肌酐(A/C)20～200 mg/g;显著蛋白尿患者(DN 2组)29例,A/C＞200 mg/g.选取正常对照组32例,均为健康体检者.所有检测对象过夜禁食10～12小时后,于清晨空腹抽取肘静脉血4 ml,其中2 ml不抗凝血用于生化指标检测.酶联免疫吸附试验(ELISA)测定血浆PAI-1、TGF-β水平.结果 ①血浆PAI-1水平DN1、DN2组显著高于正常对照组,(69.28±18.61) ng/L、(69.43±17.86) ng/L vs (51.97±30.11) ng/L(P＜0.05).②血清TGF-β水平DM、DN1、DN2组显著高于正常对照组,分别为(137.99±21.47) ng/L、(180.36±40.45) ng/L、(298.92±77.37) ng/L vs(100.65±24.21) ng/L(均P＜0.01).③血清TGF-β和血浆PAI-1水平无明显相关性.④血浆PAI-1水平及血清TGF-β水平升高是T2DM并发肾病的危险因素.结论 T2DM合并肾病患者血浆PAI-1水平、血清TGF-β水平升高,两项指标可以预测T2DM合并肾病的危险.     

含肝细胞生长因子水凝胶修复家兔股骨颈部缺损

背景：肝细胞生长因子可促进骨组织再生,在骨组织修复方面有着巨大的潜力,但是在体内较短的半衰期限制了其在临床的应用。目的：观察含有肝细胞生长因子的半合成细胞外基质样水凝胶体内促进股骨颈骨缺损修复的作用。方法：取12只兔制作双侧股骨颈骨缺损模型,采用自体配对对比,左侧为对照侧不对骨缺损做任何处置,右侧为实验侧植入含有肝细胞生长因子的半合成细胞外基质样水凝胶。结果与结论：苏木精-伊红染色见2周时实验侧缺损修复区血管分布较对照侧均匀;4周时实验侧缺损区均匀填充新生骨小梁,而对照侧由外向内骨小梁形成减少;8周时实验侧缺损区皮质骨形成,并且骨髓腔再通,有骨髓细胞填充,对照侧仍有粗大骨痂填充缺损区,堵塞髓腔。钼靶X射线观测到8周时,实验侧难以区分正常骨质与新生骨质交界线,而对照侧缺损区与周围骨质分界明显。证实含有肝细胞生长因子的半合成细胞外基质样水凝胶可以促进家兔股骨颈部骨缺损的骨组织修复。