RESUMEN
Hepatocyte growth factor (HGF) has been shown to induce angiogenesis in vivo and has potential as a candidate gene for 'therapeutic angiogenesis'. In vivo, two isoforms of HGF, HGF723 and HGF728, consisting of 723 and 728 amino acids, are generated through alternative splicing between exons 4 and 5, but the biological effects of their coexpression have not yet been elucidated. In this study, we generated a series of genomic-complementary DNA (cDNA) hybrids of the HGF gene by inserting various truncated intron 4 into the junction of exons 4 and 5 of HGF cDNA and analyzed the biological activities of these hybrid constructs. We showed that: (1) the hybrid called HGF-X7, which contained 1502 base pairs of intron 4, could drive a higher level of HGF expression than other hybrid constructs and cDNAs of each isoform alone; (2) the pCK vector was most efficient for the gene expression of HGF-X7; (3) coexpression of both isoforms of HGF could more efficiently induce the migration of human umbilical vein endothelial cell (HUVEC) and of the mouse myoblast cell line C2C12 myoblasts than a single isoform of HGF and human vascular endothelial growth factor (VEGF)165 at a given concentration; (4) intramuscular administration of pCK-HGF-X7 resulted in transient and localized HGF expression in the injected muscle without an increase in the HGF protein levels in other tissues including serum; and (5) intramuscular injection of pCK-HGF-X7 could more efficiently increase the number of angiographically recognizable collateral vessels, as well as improve an intra-arterial Doppler wire-measured blood flow in the rabbit model of hindlimb ischemia when compared with the identical vector encoding VEGF165 gene. These results showed that transfer of the genomic-cDNA hybrid of the HGF gene could be used as a potential therapeutic approach to human vascular diseases.
Asunto(s)
Arterias , Circulación Colateral/efectos de los fármacos , ADN/uso terapéutico , Terapia Genética , Factor de Crecimiento de Hepatocito/genética , Factor de Crecimiento de Hepatocito/metabolismo , Isquemia/terapia , Animales , Arterias/crecimiento & desarrollo , Arterias/metabolismo , Línea Celular , Movimiento Celular/efectos de los fármacos , ADN/genética , ADN Complementario/genética , Modelos Animales de Enfermedad , Extremidades/irrigación sanguínea , Femenino , Expresión Génica , Técnicas de Transferencia de Gen , Ingeniería Genética , Vectores Genéticos/genética , Factor de Crecimiento de Hepatocito/farmacología , Humanos , Intrones/genética , Isquemia/fisiopatología , Masculino , Ratones , Ratones Endogámicos BALB C , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Conejos , Flujo Sanguíneo Regional/efectos de los fármacosRESUMEN
Several shorter-term alternatives for whole-day ambulatory monitoring of blood pressure using Pressurometer III or conventional sphygmomanometer were evaluated in 12 male patients with mild hypertension. Averages of BP reading at 8 AM once, 3 consecutive-readings either with Pressurometer or manually, serial readings for 2-hour intervals from 8-10 AM and 2-4 PM were compared with that of 24-hour ambulatory, non-invasive BP readings by Pressurometer. Both systolic and diastolic 2-hour BP averages in the morning (8 to 10 o'clock) correlated more strongly with 24-hour averages (r = 0.91 and 0.91) than the 3 consecutive (r = 0.88 and 0.66) or single (r = 0.49 and -0.35) reading alternatives did. In conclusion, the average of serial readings obtained during 2-hour monitoring period from 8 to 10 AM is a reliable predictor of 24-hour ambulatory BP and represents it more closely than the conventional single or multiple BP readings.