肝细胞生长因子在心力衰竭中的研究进展

肝细胞生长因子具有潜在的多种生物学活性,包括致有丝分裂作用、形态形成作用、造血作用、心肌肥大、血管生成、抗凋亡、抗纤维化及组织再生作用。这将对心力衰竭产生保护作用。现就其在心力衰竭中作用及其机制进行探讨。     

Gene transfer of hepatocyte growth factor to subarachnoid space in cerebral hypoperfusion model

Although cerebral hypoperfusion caused by cerebral occlusive disease leads to cerebral ischemic events, an effective treatment has not yet been established. Recently, a novel therapeutic strategy for ischemic disease using angiogenic growth factors to expedite and/or augment collateral artery development has been proposed. Therapeutic angiogenesis might be useful for the treatment of cerebral occlusive disease. Hepatocyte growth factor (HGF) is a potent angiogenic factor, in addition to vascular endothelial growth factor (VEGF), whereas in the nervous system HGF also acts as neurotrophic factor. Therefore, we hypothesized that gene transfer of these angiogenic growth factors could induce angiogenesis, thus providing an effective therapy for cerebral hypoperfusion or stroke. In this study, we employed a highly efficient gene transfer method, the viral envelop (Hemagglutinating Virus of Japan [HVJ]-liposome) method, because we previously documented that beta-galactosidase gene could be transfected into the brain by the HVJ-liposome method. Indeed, we confirmed wide distribution of transgene expression using beta-galactosidase via injection into the subarachnoid space. Of importance, transfection of HGF or VEGF gene into the subarachnoid space 7 days before occlusion induced angiogenesis on the brain surface as assessed by alkaline phosphatase staining (P<0.01). In addition, significant improvement of cerebral blood flow (CBF) was observed by laser Doppler imaging (LDI) 7 days after occlusion (P<0.01). Unexpectedly, transfection of HGF or VEGF gene into the subarachnoid space immediately after occlusion of the bilateral carotid arteries also induced angiogenesis on the brain surface and had a significant protective effect on the impairment of CBF by carotid occlusion (P<0.01). Interestingly, coinjection of recombinant HGF with HGF gene transfer revealed a further increase in CBF (P<0.01). Here, we demonstrated successful therapeutic angiogenesis using HGF or VEGF gene transfer into the subarachnoid space to improve cerebral hypoperfusion, thus providing a new therapeutic strategy for cerebral ischemic disease.     

Angiotensin II Receptor Blockade in Syrian Hamster (T0-2) Cardiomyopathy Does Not Affect Microscopic Cardiac Material Properties: Implications for Mechanisms of Tissue Remodeling

This study delineates the role of angiotensin II type I (AT1) receptor in the remodeling of Syrian cardiomyopathic hamsters. Twelve cardiomyopathic (T0-2) hamsters received L-158,809 treatment ad libitum in their drinking water (27 μg/ml) and 9 cardiomyopathic and 9 normal F1-B hamsters received tap water from 1 to 4 months of age. Although pharmacologically effective with regard to complete suppression of the blood pressure response to angiotensin II infusion, L-158,809 did not diminish the progression or severity of cardiomyopathy. Heart weight/100 g body weight and left ventricular wall thickness adjusted for body weight of both L-158,809 and cardiomyopathic control hamsters did not differ and exceeded those of F1-B controls (p < 0.05). Myocardial material properties (e.g., stiffness and density) of cardiomyopathic hamsters treated with L-158,809 were not affected. Thus, the progression of fibrosis, calcification, and necrosis in T0-2 cardiomyopathic hamsters was not sensitive to AT1 receptor blockade. This revised version was published online in July 2006 with corrections to the Cover Date.     

Augmentation of beta adrenergic receptors in cardiomyopathic hamsters (BIO 14.6) with heart failure.

OBJECTIVE: The aim was to characterise the transmural distribution of beta adrenergic receptors in failing myocardium in cardiomyopathy. METHODS: Using a quantitative autoradiographic technique with 125I-cyanopindolol (ICYP), the density and transmural distribution of beta adrenergic receptors were compared between eight cardiomyopathic BIO 14.6 Syrian hamsters with heart failure and six normal age matched controls (BIO 14.6HAM). RESULTS: Binding of ICYP to transmural slices of hamster myocardium was rapid, saturable, stereoselective, and displaceable by antagonists. The binding isotherm showed a significant increase in the total tissue content of beta adrenergic receptors in the failing myocardium of cardiomyopathic hamsters: 15.3(SEM 1.6) fmol.mg-1 protein v 9.4(1.2) fmol.mg-1 protein in normal myocardium of control hamsters (p < 0.05). There was no difference in receptor affinity. Quantitative autoradiography showed regional heterogeneity of beta adrenergic receptors in cardiomyopathic hamsters, with an increase of beta adrenergic receptor density in the septal and subendocardial regions. In addition, the regions with increased interstitial fibrosis corresponded to the sites of increased beta adrenergic receptor density. CONCLUSIONS: The transmural distribution of beta adrenergic receptor is heterogeneous in the failing myocardium of cardiomyopathic hamsters and an increased beta adrenergic receptor density may be associated with the development of cardiomyopathy.     

Tissue-cultured heart cells from the cardiomyopathic hamster

Cultured embryonic heart cells of normal hamsters and of Syrian hamsters with hereditary idiopathic cardiomyopathy were studied. Phase contrast microscopy showed that the beat frequency decreased more rapidly and the regularity of the rhythm of the beating cells was lost sooner in the heart cells from cardiomyopathic hamsters than in control animals. Electron microscopic study of the cultured heart cells revealed a significant impediment in the maturation of the sarcomeric units in the cardiomyopathic hamsters compared to control animals. Relatively abundant corpuscles resembling neurosecretory granules were found in the paranuclear area of the cultured embryonic heart cells of the hamsters with hereditary idiopathic cardiomyopathy and it is suggested that the corpuscles might be related to a disturbed function in the cultured heart cell of the cardiomyopathic hamster.     

Treatment of dilated cardiomyopathy with electroporation of hepatocyte growth factor gene into skeletal muscle

Hepatocyte growth factor (HGF) is a potent angiogenic and antifibrotic factor. Cardioprotective effects of HGF for idiopathic dilated cardiomyopathy were examined in hamsters with electroporation of plasmid DNA into skeletal muscle. We used hamster skeletal muscle as a protein producer of HGF gene. A plasmid vector encoding HGF (HGF group, n=12) or empty plasmid (placebo group, n=12) was transferred with in vivo electroporation into tibialis anterior muscles of hamsters with inherited dilated cardiomyopathy (TO-2 strain). The HGF group had greater serum HGF levels (21.6+/-2.2 versus 0.11+/-0.07 ng/mL, P<0.05), higher left ventricular ejection fraction (47.9+/-9.4% versus 28.8+/-11.2%, P<0.05), and greater wall thickening (31.6+/-6.3% versus 19.7+/-6.1%, P<0.05) when compared with the placebo group. The HGF group had smaller areas of ventricular fibrosis (11.8+/-3.4% versus 17.1+/-3.5%, P<0.05) and lower hydroxyproline content (3.7+/-0.7 versus 5.1+/-0.9 micromol/g, P<0.05) than did the placebo group. The HGF group also had higher capillary density (1885+/-232 versus 1447+/-182 vessel/mm(2), P<0.05) and higher matrix metalloprotease-1 activity (13.1+/-3.5 versus 8.1+/-3.6 microg/collagen degraded per hour per gram tissue, P<0.05) than did the placebo group. Exogenous HGF might improve the deleterious changes in myocardial function and structure in the hamster with dilated cardiomyopathy. Systemic delivery of gene products with in vivo electroporation into skeletal muscle seemed to be an alternative means of direct gene delivery.     

Effect of chronic AT(1) receptor blockade on cardiac Smad overexpression in hereditary cardiomyopathic hamsters

OBJECTIVE: As the pharmacological suppression of angiotensin has been associated with cardioprotective effects in cardiomyopathy, our primary aim was to determine whether the expression of Smad protein components of the cardiac TGF-beta signaling cascade is modulated by chronic AT(1) receptor blockade. Furthermore, we examined the relationship between cardiac Smad protein expression and altered collagen turnover in the cardiomyopathic heart. METHODS: Male UM-X7. 1 cardiomyopathic (CMP) Syrian hamsters at early (65 days) and late (200 days) stages of cardiomyopathy were subjected to 4 week losartan (15 mg/kg/day) treatment. Expression of left ventricular (LV) receptor-activated (Smad 2) and common-mediator (Smad 4) Smads from control (F1-beta strain) hamsters, non-treated cardiomyopathic (CMP), and losartan-treated CMP animals was assessed. Collagen turnover, including fibrillar collagen synthesis/accretion and cardiac MMP activity was assessed. RESULTS: Elevated mRNA abundance of fibrillar collagens and ANF were present in cardiomyopathic hearts and these trends were normalized in the early stage losartan-treated group. 4-Hydroxyproline and zymographic assays confirmed fibrosis and elevated MMP-1 and -2 activities in CMP hearts. Losartan treatment was associated with a modest reduction of cardiac 4-hydroxyproline concentration, and a significant reduction of both MMP-1 and MMP-2 activities. While TGF-beta(1) mRNAs were elevated in both CMP groups vs. controls, total TGF-beta protein content was not different in CMP vs. controls. In LV preparations containing nuclear extract, elevated Smad 2 and Smad 4 protein expression was noted in cardiomyopathic hearts vs. controls. Losartan treatment of late-stage CMP hamsters was associated with a significant reduction in Smad 2 and a modest reduction of Smad 4 protein expression vs. untreated CMP samples. CONCLUSIONS: Altered cardiac Smad expression, present in both early and late stage cardiomyopathy, is positively correlated with the occurrence of cardiac fibrosis and elevated collagen turnover in failing CMP hearts. Four week AT(1) blockade is associated with normalized expression of cardiac Smad 2 proteins, and these changes occur in parallel with some aspects of collagen turnover in failing cardiomyopathic hearts.     

Glomerular and vascular atrial natriuretic factor receptors in cardiomyopathic hamsters: correlation with the peptide biological effects

STUDY OBJECTIVE--The aim was to study glomerular and vascular atrial natriuretic factor (ANF) receptors and their relationship with the post-receptor effects of the peptide in experimental heart failure. DESIGN--Binding sites ANF were studied in renal glomerular and mesenteric artery membranes. The natriuretic and relaxing effects of ANF were evaluated in the intact animal and in noradrenaline precontracted aortic strips respectively. Plasma and tissue ANF levels were also assessed. EXPERIMENTAL MATERIAL--The study was performed on cardiomyopathic (UM-X7.1) hamsters (n = 15) with a moderate degree of heart failure. Age matched Golden Syriam hamsters (n = 15) were used as controls. MEASUREMENTS AND MAIN RESULTS--Cardiomyopathic hamsters presented lower blood pressure, body weight, and plasma Na+, and higher heart weight than normal hamsters. Plasma ANF (1-98) and (99-126) levels and ventricular ANF content were higher in cardiomyopathic hamster than in controls. ANF and frusemide decreased blood pressure, and increased diuresis and natriuresis in normal hamsters. The blood pressure reduction by ANF in cardiomyopathic hamsters was approximately of the same magnitude as in normal hamsters but their renal response was blunted. The blood pressure lowering effect of frusemide was similar in both cardiomyopathic and normal hamsters, but the diuretic and natriuretic responses were greatly reduced in the former. Glomerular ANF receptor density was higher and receptor affinity was lower in cardiomyopathic hamsters than in controls. Noradrenaline precontracted vascular strips from cardiomyopathic hamster were more sensitive to the relaxant effect of ANF than those from controls. No differences in either density or affinity of vascular receptor were observed. CONCLUSIONS--The results suggest that the renal hyporesponsiveness of cardiomyopathic hamsters to ANF is not due to a down regulation of glomerular ANF receptors. The fact that the natriuretic response to frusemide is also blunted suggest the involvement of other factors.     

Cellular oncogene expression in the idiopathic cardiomyopathic hamster heart during the growing process

The expression of various proto-oncogenes was evaluated in the Syrian hamster with hereditary idiopathic cardiomyopathy. mRNA from the heart and aorta of controls (BIO-RB) and cardiomyopathic hamsters (UM-X7.1 strain, BIO 14.6 line) was tested using RNA hybridization techniques. Of the 19 different v-oncogene probes used in the study, only the v-myc probe revealed a substantially greater expression of oncogene in the 30th day of cardiomyopathic hamster heart than in control hamster heart. The amplified expression of c-myc was also observed in the heart of 1-year-old, but not of 7-day-old cardiomyopathic hamster. Overexpression of c-myc, otherwise associated with the regulation of cell differentiation or rapid growth, may relate to the pathological state or pathogenesis of the hereditary cardiomyopathy.     

Effects of verapamil on experimental cardiomyopathy in the Bio 14.6 Syrian hamster

The Bio 14.6 Syrian hamster provides a good model for experimental study of cardiomyopathy. Cardiac receptor binding sites (alpha-1-, beta- and calcium antagonists) were studied in early (21 days old) and late (70 days old) stages of cardiomyopathy. The effects of verapamil on histologic features and free radicals in the heart were studied. The number of alpha-1- and beta-cardiac receptor binding sites was significantly greater in the late stage of cardiomyopathy when compared with findings in normal golden hamsters used as controls. The calcium antagonist receptors were significantly increased in the early stage but alpha-1- and beta-receptors were not. Verapamil-treated hamsters received intraperitoneal injections of verapamil at a dose of 5 mg/kg per day for 70 days from age 20 days. The percent areas of fibrosis and calcification in the verapamil-treated group were significantly smaller than those in the control group. The concentrations of lipid peroxides in the whole heart and free radicals in the heart mitochondria were significantly higher in the cardiomyopathic hamsters, and verapamil inhibited the increase in free radical concentration in the hearts of these hamsters. This study confirms that the number of calcium channels is increased early in the course of cardiomyopathy in the Bio 14.6 Syrian hamster. A larger number of free radicals may participate in the accumulation of calcium and cell injury in the myocytes of these hamsters. Verapamil protects against myocardial damage and may do so by inhibiting voltage-dependent calcium uptake and by preventing cell injury from free radicals.     

Cardiac adaptation and its limitation in an experimental model of congestive heart failure.

To elucidate mechanisms of adaptation and maladaptation in heart failure, abnormalities of left ventricular function and their relationships to myocardial contractile protein were studied in the Syrian hamster Bio 14.6. Left ventricular and heart weights were both increased in 20-week-old cardiomyopathic hamsters, indicating cardiac hypertrophy as a compensatory mechanism to the disease process of cardiomyopathy. However further increase in the left ventricular weight was not observed in older (40-week-old) cardiomyopathic hamsters. On the other hand left ventricular volume and volume/mass ratio were increased progressively. Correspondingly, V3 type myosin was increased and myosin sliding velocity was decreased. Left ventricular function of cardiomyopathic hamsters evaluated using an isovolumically beating perfused heart preparation was depressed, and this functional impairment was also progressive. Chronic administration of metoprolol, a beta-blocking agent, induced further increase in left ventricular volume and mass without changing left ventricular function and myosin isozyme pattern. Thus in cardiomyopathic hamsters, left ventricular function progressively deteriorates in spite of a variety of adaptive mechanisms, and remodeling occurs.     

Angiotensins and the failing heart. Enhanced positive inotropic response to angiotensin I in cardiomyopathic hamster heart in the presence of captopril

We examined the hypothesis that the positive inotropic effect of angiotensin I (Ang I) may be retained in the presence of angiotensin converting enzyme inhibitors so that it may have a direct beneficial effect on the heart. Accordingly, isolated perfused hearts (Langendorff preparation) of 300-day-old cardiomyopathic hamsters (a model of spontaneous cardiomyopathy) and age-matched normal hamsters (controls) were infused with Ang I in the presence of captopril; propranolol was added to the perfusing medium to block catecholamine-mediated effects of angiotensins on the heart. Left ventricular developed pressure and the rate of increase in left ventricular developed pressure increased significantly (p less than 0.001) in both the cardiomyopathic and the normal hamster heart despite concomitant reduction in myocardial flow rate favoring a direct inotropic effect of Ang I in both normal and myopathic hearts; these changes were significantly higher by almost threefold in the cardiomyopathic than in the normal hamsters (p less than 0.01) and were blocked by the angiotensin II (Ang II) antagonist [Sar1,Thr8]Ang II. Comparing dose-left ventricular contractility response curves for Ang I and Ang II, ED50 for responses was identical in both normal and myopathic hearts, whereas peak responses to Ang II were double those to Ang I in normal hearts but were almost identical in the myopathic hearts. Binding of [125I]Ang II in six cardiomyopathic and four normal hamster hearts was of high affinity, but there was no evidence for Ang I-saturable high-affinity binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)     

Does an impaired adenosine mediated feedback control play a role in the development of hereditary dystrophic cardiomyopathy?

A study was carried out to investigate whether or not an impairment of the adenosine mediated negative inotropic effect in the presence of beta adrenoceptor stimulation plays a role in the pathogenesis of the hereditary cardiomyopathy of the Syrian hamster. In electrically driven papillary muscles isolated from the hearts of cardiomyopathic (strain BIO 8262) and age matched healthy control Syrian hamsters the effects of isoprenaline, adenosine, and adenosine in the presence of isoprenaline were studied within the first 30 days of life (the prenecrotic stage of the disorder). In both cardiomyopathic and control hamsters adenosine antagonised the positive inotropic effect of isoprenaline, whereas adenosine alone had no or, only a weak, inhibitory effect on the force of contraction. The effects in both groups were similar. The effect of isoprenaline on the force of contraction also did not differ in the two groups. The data show that in both cardiomyopathic and control hamsters adenosine reduces the force of contraction during beta adrenergic stimulation. The potency or efficacy of adenosine did not differ in the two groups. An impaired adenosine mediated feedback control of the heart does not therefore seem to play a role in the pathogenesis of the hereditary dystrophic cardiomyopathy of the Syrian hamster.     

VEGF-mediated angiogenesis is impaired by angiotensin type 1 receptor blockade in cardiomyopathic hamster hearts

OBJECTIVE: Coronary microcirculation plays an important role in the progression of cardiac remodeling. Among angiogenic factors, it has been reported that angiotensin II may contribute to neovascularization. However, it is unknown whether inhibition of the renin-angiotensin system suppresses angiogenesis, especially within the heart. Our aim was to evaluate the effects of the angiotensin-converting enzyme inhibitor enalapril and the angiotensin II receptor type I blocker valsartan on cardiac microvasculature, function, vascular endothelial growth factor (VEGF) expression, and survival in cardiomyopathic hamsters. METHODS: Male cardiomyopathic hamsters (BIO TO2) were administered either a placebo (group C), enalapril (30 mg/kg/day) (group E), or valsartan (40 mg/kg/day) (group V), starting at the age of 6 weeks. This continued until death. Hemodynamic study, histological analysis, and northern blot analysis were performed at 39 weeks. RESULTS: Group V showed significant increases in percent fibrosis, end diastolic pressure, and LV dP/dt min, and significant decreases in percent fractional shortening, LV dP/dt max, capillary density, and the level of mRNA expression of VEGF compared with group C. Group E showed significant increases in percent fractional shortening while the capillary density and level of mRNA expression of VEGF were unchanged. The 300-day survival rate was significantly lower in group V (25.0%) but higher in group E (100%) than that of group C (66.7%). CONCLUSIONS: Therapy with valsartan may have adverse effects on survival rate concomitant with the progression of cardiac remodeling owing to impaired VEGF-mediated angiogenesis. Therapy with enalapril has a neutral effect on VEGF-mediated angiogenesis, leading to the suppression of cardiac remodeling and an increase in life expectancy.     

基因修饰胎肝干细胞治疗肝纤维化

目的探讨肝细胞生长因子（HGF）基因修饰的胎肝干细胞治疗肝纤维化大鼠的效果。方法构建大鼠HGF基因和绿色荧光蛋白基因共同表达的融合载体,转染到胎肝母细胞中。将能够表达HGF的细胞移植到CCl4肝纤维化大鼠体内,将增强型绿色荧光蛋白（EGFP）修饰的胎肝干细胞作为对照组,检测大鼠肝功能和血清HGF及TGFβ1的浓度,肝组织中荧光蛋白数量和分布,肝组织内Ⅰ、Ⅲ型胶原蛋白的沉积情况。结果组织学及血清学检测显示,与对照组相比,经HGF基因修饰的胎肝干细胞能够显著改善肝纤维化指标和大鼠肝功能（P〈0.05）。结论 HGF修饰的胎肝干细胞治疗可能成为肝纤维化潜在的治疗手段。     

Possible involvement of free radicals and antioxidants in the early stages of the development of cardiomyopathy in BIO 14.6 Syrian Hamster.

The BIO 14.6 cardiomyopathic Syrian hamster is a well-known animal model of congestive cardiomyopathy. To evaluate the role of free radicals and antioxidant protection in the pathogenesis of cardiomyopathy in this animal, we studied the concentration of heart mitochondrial free radicals, the activities of glutathione peroxidase (GSHPx) and superoxide dismutase (SOD), and the effect of alpha-tocopherol on the early stage of myocardial damage (up to 90 days). The GSHPx activity in BIO 14.6 hamsters was found to be twice that in the normal control hamsters at 30 days of age, while SOD activity was unchanged at 30 and 90 days of age. The concentrations of mitochondrial free radicals in BIO 14.6 hamsters at 40 and 90 days of age were significantly higher than those in the normal control hamsters. A protective effect of alpha-tocopherol therapy was shown in BIO 14.6 hamsters treated during the early stage of cardiomyopathy (up to 90 days). These results show the role of free radicals and antioxidant protection in the pathogenesis of hamster cardiomyopathy. We suspect that an increase in the GSHPx activity in BIO 14.6 hamsters may be due to a compensatory mechanism to counteract oxidative stress, but antioxidant reserve was not sufficient to protect the heart from the toxic effects of increased free radicals in the early stage of cardiomyopathy.     

Evaluation of the hereditary Syrian hamster cardiomyopathy by 31P nuclear magnetic resonance spectroscopy: improvement after acute verapamil therapy

The relation between metabolic and functional derangement in various cardiomyopathies has not been well characterized. This information was specifically sought in a spontaneous cardiomyopathic model. Metabolic and hemodynamic parameters were obtained in glucose-perfused beating hearts of 180-200-day-old cardiomyopathic Syrian hamsters and age-matched healthy animals. This period in the cardiomyopathic hamster lifetime is intermediary between the necrotic phase and the appearance of heart failure. We used 31P nuclear magnetic resonance spectroscopy to analyze energy metabolites and intracellular pH. Cardiomyopathic hamsters had significantly higher mole fraction values for inorganic phosphate, lower phosphocreatine mole fraction as well as lower phosphocreatine/inorganic phosphate and adenosine triphosphate/inorganic phosphate ratios. Analysis of pH indicated the presence of regions of increased acidity within the heart of myopathic hamsters. Cardiomyopathic hamsters also had significantly lower left ventricular pressure, coronary flow, and myocardial oxygen consumption. Separate groups of normal and myopathic hamsters were given verapamil for 24 hours (one injection of 4 mg/kg s.c. followed by 1.2 g/l in drinking water). Verapamil-treated myopathic hamsters had evidence of markedly improved mitochondrial function when compared with untreated animals. Left ventricular pressure and coronary flow rose to normal levels. Replacing glucose by pyruvate in the perfusate of myopathic hamsters results in a marked increase in left ventricular pressure, coronary flow, and oxygen consumption with a moderate rise in phosphocreatine. Thus, 180-200-day-old cardiomyopathic hamster heart is characterized by evidence of decreased mitochondrial function, by areas of increased acidity within the heart, and by reduced left ventricular function.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hepatocyte growth factor gene transfer with naked plasmid DNA ameliorates dimethylnitrosamine-induced liver fibrosis in rats

Aim: Hepatocyte growth factor (HGF) has various biological properties, including antifibrogenic activity. In the present study, we tested the efficacy of HGF gene therapy using naked plasmid DNA in dimethylnitrosamine (DMN)-induced liver fibrosis in a rat model. Methods: Naked plasmid DNA encoding human HGF was injected once, together with a hypertonic solution, into the hepatic artery after DMN treatment on three consecutive days per week for 3 weeks. Naked plasmid DNA encoding beta-galactosidase was injected similarly in the DMN-treated control rats. DMN treatment was continued once weekly after gene transfer for additional 3 weeks. Results: The human HGF protein expression was detected in livers transfected with human HGF naked plasmid DNA, gradually decreasing by day 21. The expression of the endogenous rat HGF protein was also upregulated after human HGF gene transfer. Phosphorylation of c-Met, a HGF receptor, was detected only in livers transfected with human HGF plasmid DNA. Fibrosis was attenuated significantly in livers transfected with the human HGF plasmid. Attenuation wasaccompanied by decreased expression of alpha-smooth muscle actin. Increased portal vein pressure after treatment with DMN was suppressed significantly by HGF gene transfer. The upregulated hepatic protein expression of transforming growth factor-beta (TGF-beta) in response to DMN was markedly attenuated by HGF gene transfer accompanied by the increased protein expression for matrix metalloproteinases (MMP)-3 and -13. Conclusion: The hepatic arterial injection of human naked plasmid HGF DNA was effective in suppressing liver fibrosis induced in rats by DMN. The mechanisms by which HGF expression attenuated liver fibrosis may include the suppression of hepatic TGF-beta expression and the induction of MMP expression.     

Hepatocyte growth factor ameliorates the progression of experimental autoimmune myocarditis: a potential role for induction of T helper 2 cytokines

Hepatocyte growth factor (HGF) plays a role in cell protection, antiapoptosis, antifibrosis, and angiogenesis. However, the role of HGF in the immune system is not well defined. We examined the influence of HGF on T cells and the effects of HGF therapy in acute myocarditis. Lewis rats were immunized on day 0 with cardiac myosin to establish experimental autoimmune myocarditis (EAM). Human HGF gene with hemagglutinating virus of the Japan-envelope vector was injected directly into the myocardium on day 0 or on day 14 (two groups of treated rats). Rats were killed on day 21. Expression of c-Met/HGF receptor in splenocytes and myocardial infiltrating cells was confirmed by immunohistochemical staining or FACS analysis. Myocarditis-affected areas were smaller in the treated rats than in control rats. Cardiac function in the treated rats was markedly improved. An antigen-specific T cell proliferation assay was done with CD4-positive T cells isolated from control rats stimulated with cardiac myosin. HGF suppressed T cell proliferation and production of IFN-gamma and increased production of IL-4 and IL-10 secreted from CD4-positive T cells in vitro. Additionally, TUNEL assay revealed that HGF reduced apoptosis in cardiomyocytes. HGF reduced the severity of EAM by inducing T helper 2 cytokines and suppressing apoptosis of cardiomyocytes. HGF has potential as a new therapy for myocarditis.