Hepatocyte Growth Factor Genetic Variations and Primary Angle-Closure Glaucoma in the Han Chinese Population

Purpose

The aim of this study is to examine whether or not hepatocyte growth factor (HGF) genetic variations are associated with susceptibility to primary angle-closure glaucoma (PACG) in the Han Chinese population.

Methods

Three single-nucleotide polymorphisms (SNPs)–rs5745718, rs17427817, and rs3735520–in the HGF gene were genotyped in 238 adult patients with PACG and 287 age-, sex-, and ethnically matched healthy controls by using a polymerase chain reaction restriction fragment length polymorphism assay. Data was analyzed by χ² analysis.

Results

The three tested analyzed polymorphisms in the HGF gene were in Hardy-Weinberg equilibrium, in all the subjects. The frequencies of the genotype and allele of rs5745718 and rs1742817 in the HGF gene were significantly different between the PACG patients and the controls. On one hand, the frequencies of the CC genotype and C allele of rs5745718 were significantly decreased in PACG patients compared with controls (P_c = 1.40×10⁻³; P_c = 3.21×10⁻⁴, respectively); however, on the other hand, significantly decreased frequencies of the GG genotype and the G allele of rs17427817 were observed in PACG patients compared with the controls (P_c = 0.006,; P_c = 6.06×10⁻⁴, respectively). A comparison of the distributions of the genotypes and alleles of rs3735520 showed no statistically significant differences between the PACG patients and the controls (p_c>0.05). The haplotype analysis results showed that the CGC haplotype frequency was significantly decreased in the patients with PACG compared with the controls (p_c<0.001). No difference was detected between the patients and the controls with regard to the other haplotypes.

Conclusions

Our study suggests that rs5745718 and rs17427817 are associated with a decreased risk of PACG in the Chinese Han population. The CGC haplotype was demonstrated to possibly play a protective role against PACG in this population.     

肝细胞生长因子抗肾纤维化研究进展

慢性肾脏疾病患者的肾功能会随时间的推移而进行性恶化,肾实质细胞进行性丧失及细胞外基质蛋白过度沉积将导致肾纤维化形成,肾纤维化进行性发展将最终走向终末期肾衰竭。肝细胞生长因子(HGF)及其受体c-Met对肾发育和急性肾损伤后的肾脏再生修复具有重要作用,在慢性肾衰竭及肾纤维化时,HGF还具有营养肾脏及抗肾纤维化的作用。简要综述了HGF抑制肾纤维化形成的细胞分子机制的研究进展,提示HGF在治疗肾纤维化方面所具有的前景。     

携带人肝细胞生长因子重组质粒pUDK-HGF对大鼠肾纤维化的防治作用

目的:研究肝细胞生长因子(HGF)基因转导对庆大霉素诱导的大鼠肾纤维化损伤的防治效果。方法:以雄性Wistar大鼠腹腔注射硫酸庆大霉素注射液制备肾纤维化模型;实验分为正常对照组、肾纤维化模型组、HGF治疗组;造模后第30 d,HGF治疗组于左侧肾脏直接注射重组质粒pUDK-HGF注射液,模型组注射质粒pUDK,正常对照组只进行假手术;于造模后第60 d处死大鼠,评价HGF对血尿素氮、血肌酐、24 h尿蛋白、肾系数等肾功能指标的改善作用,并对肾纤维化进行组织学评价。结果:与正常对照组相比,模型组肾功能下降,肾系数(8.8±0.95 g/kg)、血尿素氮(9.4±2.61 mmol/L)、血肌酐(42±10.33μmol/L,P<0.05)及24 h尿蛋白定量(25.78±8.66 mg,P<0.05)升高;HGF治疗组对肾功能有所改善,可缓解肾纤维化的进展。此外,本实验表明,对已纤维化肾脏直接注射HGF基因,可促进肾间质血管再生,并进一步降低肾小管间质损伤积分。结论:将HGF基因靶向导入大鼠体内可有效防治肾纤维化。     

肝细胞生长因子:一种多功能细胞因子

肝细胞生长因子（ｈｅｐａｔｏｃｙｔｅｇｒｏｗｔｈｆａｃｔｏｒ,ＨＧＦ）,又名离散因子（ｓｃａｔｅｒｆａｃｔｏｒ）。其受体ｃ－ＭＥＴ是原癌基因ｃ－ｍｅｔ的产物。ＨＧＦ和ｃ－ＭＥＴ在多种细胞中都有产生,组织分布广泛。ＨＧＦ具有器官再生、创伤愈合、胚胎形成...     

肝细胞生长因子在神经系统的研究进展

肝细胞生长因子是一多效性细胞因子。在神经系统发育和再生过程中,肝细胞生长因子作为神经营养因子发挥作用。本文就肝细胞生长因子的分子生物学特性以及在神经系统中的分布和生物学作用进行综述。     

肝细胞生长因子在骨骼肌再生中的作用研究进展

骨骼肌损伤后的修复包括炎症反应期、修复期、组织重塑期三个阶段。而骨骼肌卫星细胞的激活、增殖与分化和骨骼肌伤后的修复有着密切的关系。骨骼肌损伤后,肝细胞生长因子(HGF)可以自分泌、旁分泌或内分泌的形式,调控肌卫星细胞功能,从而影响损伤骨骼肌的再生。其机制研究表明,HGF可能通过与其受体c-met结合,启动相关信号途径,参与骨骼肌卫星细胞激活、增殖、分化和迁移,从而影响骨骼肌再生进程。     

壳聚糖纳米基因载体介导IL-10和HGF基因重组体在大鼠活体肝移植中的研究

目的：研究hIL-10和HGF重组体对大鼠活体肝移植术后急性排斥反应的抑制作用。方法：以DA大鼠（RT1a）为供体．Lewis大鼠（RT11）为受体建立异种肝移植鼠模型,实验组于移植后1天及7天,腹腔内注射T—HGF和T-hIL-10重组体（100μg／ml）100μl,对照组注射等量生理盐水。移植后1,7,10,20天取大鼠尾静脉血,ELISA法测定血清HGF、IL-10水平,RT-PCR测定肝细胞HGF、IL-10m RNA的表达水平,常规肝功能测定,活体肝移植动物生存期观测。结果：实验组鼠肝细胞有HGF和IL-10 mRNA表达,对照组肝细胞未见表达。对照组大鼠HGF和IL-10水平一直维持在较低水平,实验组大鼠HGF和IL-10水平明显高于对照组（P〈0．05）。注射重组体的实验组平均生存时间为（19±0．9）天,20天时存活只数为6只,存活率75％;对照组平均生存时间为（8．6±0．5）天,20天时存活只数为2只,存活率75％（P〈0．05）。对照组未注射重组体,ALT和AST值持续升高,注射重组体后ALT和AST值亦有升高,但明显低于对照组（p〈0．05）。结论：活体肝移植大鼠导入IL—10和HGF基因重组体后可抑制移植术后急性排斥反应．     

肝细胞生长因子在损伤肾组织中的作用

肝细胞生长因子（hepatocyte growth factor．HGF）是一种多效性生长因子,主要由间质细胞产生,通过自分泌和旁分泌方式作用于上皮细胞、内皮细胞以及间质细胞本身,具有促有丝分裂、促细胞形态形成和调节细胞活动的功能,从而对损伤的器官和组织进行修复。许多新的研究显示,在急性肾损伤时给予外源性HGF可以保护肾小管上皮细胞、重建肾小管结构和维持肾功能完整性。此外,HGF还能有效地抑制与慢性肾脏疾病及慢性肾功能衰竭密切相关的肾间质纤维化的进展过程。     

肝细胞生长因子基因转染对人淋巴瘤细胞凋亡的影响

探讨肝细胞生长因子（HGF）基因转染人淋巴瘤细胞系Raji细胞后,拮抗足叶乙甙（VP－16）诱导细胞凋亡的研究。将三种细胞：未转染Raji细胞、空载体pVITR02－mcs转染细胞和HGF基因转染细胞,分成正常对照组和经vP－16处理的药物组。采用Westernblot法验证HGF蛋白的表达：CCK－8法检测诱导Raji细胞凋亡的药物浓度;通过透射电镜、流式细胞术、吖啶橙（A0）染色、苏木精咿红（HE）染色等方法观察Raji细胞的凋亡情况,并进行相关分析。结果显示：Westernblot法验证了HGF蛋白质的表达;CCK．8法显示100μg／mL足叶乙甙可明显抑制Raii细胞增殖;透射电镜下可发现典型的凋亡细胞;流式检测结果表明：给药组与正常组相比,三组细胞的凋亡率明显升高（P〈0．01）,提示VP－16具有诱导细胞凋亡的作用：但给药组间：HGF基因转染组凋亡率明显低于未转染组（P＜0．05）和空载体pVITR02．mcs转染组（P〈0．05）,提示嬲F基因转染可明显抑制VP-16诱导的Raji细胞的凋亡,AO染色和HE染色结果也同样提示HGF具有拮抗VP－16诱导的细胞凋亡效应。     

壳聚糖纳米基冈载体介导IL-10和HGF基因重组体在大鼠活体肝移植中的研究

目的:研究hIL-10和HGF重组体对大鼠活体肝移植术后急性排斥反应的抑制作用.方法:以DA大鼠(RTla)为供体,Lewis大鼠(RTll)为受体建立异种肝移植鼠模型,实验组于移植后1天及7天,腹腔内注射T-HGF和T-hIL-10重组体(100μg/ml)100μl,对照组注射等量生理盐水.移植后1,7,10,20天取大鼠尾静脉血,ELISA法测定血清HGF、IL-10水平,RT-PCR测定肝细胞HGF、IL-10mRNA的表达水平,常规肝功能测定,活体肝移植动物生存期观测.结果:实验组鼠肝细胞有HGF和IL-10mRNA表达,对照组肝细胞未见表达.对照组大鼠HGF和IL-10水平一直维持在较低水平,实验组大鼠HGF和IL-10水平明显高于对照组(P＜0.05).注射重组体的实验组平均生存时间为(19+0.9)天,20天时存活只数为6只.存活率75%;对照组平均生存时间为(8.6±0.5)天,20天时存活只数为2只,存活率75%(P＜0.05).对照组未注射重组体,ALT和AST值持续升高,注射重组体后ALT和AST值亦有升高,但明显低于对照组(p＜0.05).结论:活体肝移植大鼠导入IL-10和HGF基因重组体后可抑制移植术后急性排斥反应.     

不同浓度HGF、EGF优化大鼠胎肝干细胞体外培养的研究

目的:研究不同浓度肝细胞生长因子(Hepatocyte growth factor,HGF)和表皮生长因子(epidermal growth factor,EGF)对大鼠胎肝干细胞体外增殖的影响,探索二者间有无协同作用.方法:取孕14天胎龄F344大鼠胚胎的胎肝,经三步分离法分离纯化后,配置不同浓度HGF、EGF及HGF和EGF联合组培养基,将胎肝干细胞分组培养.光镜下观察细胞增殖状况,MTT法观察不同浓度和不同时间HGF、EGF及HGF和EGF联合对大鼠胎肝干细胞增殖的影响,并进行统计学分析.结果:HGF 10～80 ng/mL各浓度组,EGF 10～80 ng/mL各浓度组增殖效应均大于对照组.当HGF为20ng/mL,增殖效应明显大于对照组(P＜0.01),当HGF浓度继续增高时,增殖效应无明显增高.将20 ng/mL HGF组与不同浓度EGF组合后分组培养细胞,发现20 ng/ml HGF和10 ng/mLEGF联合组增殖效应明显升高,继续升高联合组中EGF浓度,增殖效应无明显提高.结论:HGF和EGF具有明显改善大鼠胎肝干细胞体外无血清培养条件的作用,二者对大鼠胎肝干细胞体外培养具有协同促进作用.其中20 ng/mL HGF和10 ng/mL EGF联合培养促进大鼠胎肝干细胞增殖效果最明显.     

The Aromatase Gene (CYP19A1) Variants and Circulating Hepatocyte Growth Factor in Postmenopausal Women

Background

Estrogen and androgen have been linked to the regulation of circulating hepatocyte growth factor (HGF), an adipose tissue-derived cytokine. It is possible that the CYP19A1 gene which alters sex hormones production may influence HGF levels. We examined the association between the CYP19A1 gene variants and plasma HGF concentrations.

Design

We evaluated 45 common and putative functional variants of CYP19A1 and circulating levels of HGF among 260 postmenopausal women who later developed colorectal cancer from the Women''s Health Initiative Observational Cohort. As the distribution of HGF levels was highly skewed, we transformed HGF concentrations for all women into a log-, ranked-, or normal score-scale value. Multiple linear regression with adjustment for age was used to evaluate the associations.

Results

We observed an association between the rs7172156, rs1008805, rs6493494, rs749292, and rs11636639 variants and HGF levels in ranked and normal score scales (corrected p values ≤0.02), although the association of these 5 SNPs with log-scale HGF was not significant (corrected p values ≥0.16). The associations remained unchanged after additional adjustment for hormone therapy use and estradiol levels. These 5 SNPs, which were in linkage disequilibrium (pairwise D′≥97%, r²≥56%), constituted a block with 2 common haplotypes accounting for 82% frequency. The most common haplotype, TCCCA, was associated with lower ranked- or normal score-transformed HGF levels (corrected p values ≤0.001), whereas the second most common haplotype, CTTCA, was associated with higher ranked- or normal score-transformed HGF levels (corrected p values ≤0.02).

Conclusion

Our findings of a potential association between the CYP19A1 variants and circulating HGF levels warrant confirmation in studies with larger sample size.     

肝细胞生长因子的分子生物学研究

肝细胞生长因子 (HGF)是一种多功能细胞因子 ,其分子为异二聚体糖蛋白 ,有NK1,NK2 ,NK4三个变种。HGF启动子的结构很复杂 ,其表达受多种因素和调控元件的调节。因HGF在体内有重要作用 ,HGF的基因工程表达和基因治疗正在研究中     

血管内皮细胞生长因子基因-2578C/A多态性与中国北方汉族人群银屑病易感性的相关性研究

目的:探讨血管内皮细胞生长因子(vascular endothelial cell growth factor,VEGF)基因-2578 C/A单核苷酸多态性位点与中国北方地区银屑病易感性的相关性.方法:随机收集2005年10月至2007年5月在哈尔滨医科大学附属一、二院皮肤科门诊就诊的246例寻常型银屑病患者(实验组)和271名正常对照个体(对照组)的外周静脉血,使用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法分析VEGF-2578 C/A多态位点基因型,并进行统计学分析.结果:在实验组中,VEGF-2578 C/A等位基因频率分别为74.80％、25.20％;在正常对照组中,VEGF-2578 C/A等位基因频率分别为76,59％、23.41％,两组VEGF-2578 C/A多态位点基因型的分布差异无统计学意义(P＞0.05).结论:VEGF基因-2578 C/A的多态性与中国北方汉族人群银屑病的易感性无明显相关性.     

Hepatocyte Growth Factor Signaling in Intrapancreatic Ductal Cells Drives Pancreatic Morphogenesis

In a forward genetic screen for regulators of pancreas development in zebrafish, we identified donut^s908, a mutant which exhibits failed outgrowth of the exocrine pancreas. The s908 mutation leads to a leucine to arginine substitution in the ectodomain of the hepatocyte growth factor (HGF) tyrosine kinase receptor, Met. This missense mutation impedes the proteolytic maturation of the receptor, its trafficking to the plasma membrane, and diminishes the phospho-activation of its kinase domain. Interestingly, during pancreatogenesis, met and its hgf ligands are expressed in pancreatic epithelia and mesenchyme, respectively. Although Met signaling elicits mitogenic and migratory responses in varied contexts, normal proliferation rates in donut mutant pancreata together with dysmorphic, mislocalized ductal cells suggest that met primarily functions motogenically in pancreatic tail formation. Treatment with PI3K and STAT3 inhibitors, but not with MAPK inhibitors, phenocopies the donut pancreatic defect, further indicating that Met signals through migratory pathways during pancreas development. Chimera analyses showed that Met-deficient cells were excluded from the duct, but not acinar, compartment in the pancreatic tail. Conversely, wild-type intrapancreatic duct and “tip cells” at the leading edge of the growing pancreas rescued the donut phenotype. Altogether, these results reveal a novel and essential role for HGF signaling in the intrapancreatic ducts during exocrine morphogenesis.     

Hepatocyte Growth Factor Induces Transglutaminase Activity That Negatively Regulates the Growth Signal in Primary Cultured Hepatocytes

Transglutaminase (TGase) activity increased 2.5-fold at 6 h after treatment of rat hepatocytes with 117 nMhepatocyte growth factor (HGF). In the same manner, putrescine incorporation into proteins of cells occurred in HGF-treated cells but did not in those pretreated with monodansylcadaverine (MDC), a TGase inhibitor, even in the presence of HGF. These results suggest that HGF-induced TGase was active and catalyzed some cross-linkage reaction. Cycloheximide completely blocked the increase in TGase activity induced by HGF, suggesting that HGF stimulatedde novosynthesis of TGase within 6 h. Both [³⁵S]methionine incorporation and Northern blotting analyses supported this possibility. Pretreatment of cells with MDC additionally increased HGF-induced DNA synthesis and the ratio of cells in S-phase. Similarly, TGase antisense oligonucleotide inhibitedde novosynthesis of TGase, resulting in increase in the ratio of S-phase cells in the presence of HGF. Analyses of cross-linking of HGF to the receptor indicated that the antisense oligonucleotide inhibited the downregulation of HGF receptor subsequent to HGF-addition. These results provide the first evidence for inducibility ofde novosynthesis of TGase by HGF and suggest that TGase negatively regulates the growth signal of HGF through the downregulation of receptor.     

人肝细胞生长因子cDNA克隆在CHO细胞中的表达

肝细胞生长因子是一种由α、β链组成的杂合二聚体糖蛋白,能促进肝细胞、多种上皮细胞、内皮细胞和神经胶质细胞的有丝分裂,并对多种肿瘤细胞具有细胞毒性作用或者抑制其生长［１～４］,其作用无种属特异性,如人肝细胞生长因子能促进大鼠肝细胞增殖［５］。由于天然Ｈ...     

Association of IRGM Gene Mutations with Inflammatory Bowel Disease in the Indian Population

Background

Mutations in the IRGM gene have been associated with Crohn''s disease in several populations but have not been explored in Indian patients with this disease. This study examined the association of IRGM mutations with ulcerative colitis and Crohn''s disease in Indian patients with inflammatory bowel disease.

Methods

The IRGM gene was amplified in four segments and Sanger-sequenced in 101 participants (42 Crohn''s disease, 39 ulcerative colitis, and 20 healthy controls). Ten single nucleotide polymorphisms (SNP) were genotyped in 1200 participants (352 Crohn''s disease, 400 ulcerative colitis, and 448 healthy controls) using Sequenom MassARRAY iPLEX. Disease associations were evaluated for each of the ten SNPs.

Results

Thirty one mutations were identified in the IRGM gene, of which two had not hitherto been reported (150226250- ss947429272 & 150227858- ss947429273). Ten SNPs (6 from the above and 4 from the literature) were evaluated. Significant associations with Crohn''s disease were noted with the T allele of rs1000113 (OR 1.46, 95% CI 1.12–1.90), T allele of rs9637876 (OR 1.25, 95% CI 1.005–1.561) and C allele of rs 13361189 (OR 1.33, 95% CI 1.07–1.669). Two SNPs – rs11747270 and rs180802994 – did not exhibit Hardy-Weinberg equilibrium but were associated with both Crohn''s disease and ulcerative colitis in this population. The remaining SNPs did not show significant associations with either Crohn''s disease or ulcerative colitis.

Conclusions

Association of IRGM gene SNPs with Crohn''s disease is reported for the first time in Indian patients. We also report, for the first time, an association of rs 9637876 in the IRGM gene with Crohn''s disease.