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Immunohistochemical analysis of HGF expression. HGF expression in muscle (a, d), skin (b, e) and lung (c, f) of mice treated with HGF–HVJ-liposome and BLM by using anti-human HGF antibody (a–c). The isotype control antibody used was mouse IgG antibody (d–f).
Source publication
Systemic sclerosis (SSc) is a connective tissue disorder with an unknown etiology. There are currently no effective therapies for SSc. (In this study, working with a bleomycin(BLM)-induced scleroderma model mice, we performed two transfections of human hepatocyte growth factor (HGF) cDNA into the skeletal muscle and showed that this treatment not o...
Context in source publication
Context 1
... analysis revealed a diffuse expression of HGF in the muscle, skin and lung ( Figure 3). These data indicate the transfection of the human HGF gene to mice not only in the skeletal muscle but also in the skin and lung to thus be a successful route to for achieving gene action, while HGF produced in the muscle, skin and lung is then delivered through the serum to other organs. ...
Citations
... HGF activates the c-Met receptor, thereby activating the downstream PI3K/Akt and ERK1/2 pathways, and exerts anti-inflammatory effects by downregulating NF-κB signalling, decreasing IL-6 production, increasing IL-10 production, and promoting Treg differentiation [41,42]. It also exerts anti-fibrotic effects by downregulating TGF-β signalling, suppressing fibroblasts, increasing matrix metalloproteinase 9, and decreasing tissue inhibitor of metalloproteinases 1 and 2 [43][44][45]. The efficacy of HGF on BLM-SSc mice and the enhancing anti-inflammatory and anti-fibrotic effects of BM-MSCs transfected with the HGF gene have been reported [45,46]. ...
... It also exerts anti-fibrotic effects by downregulating TGF-β signalling, suppressing fibroblasts, increasing matrix metalloproteinase 9, and decreasing tissue inhibitor of metalloproteinases 1 and 2 [43][44][45]. The efficacy of HGF on BLM-SSc mice and the enhancing anti-inflammatory and anti-fibrotic effects of BM-MSCs transfected with the HGF gene have been reported [45,46]. ...
Background
Adipose-derived mesenchymal stem cells (ASCs) have gained attention as a new treatment for systemic sclerosis (SSc). Low-molecular-weight heparin (LMWH) enhances cell function and stimulates the production of hepatocyte growth factor (HGF) in a variety of cells. This study investigated the effects of LMWH on the functions of mouse ASCs (mASCs), and the therapeutic effects of mASCs activated with LMWH (hep-mASCs) in mouse models of SSc.
Methods
The cellular functions of mASCs cultured with different concentrations of LMWH were determined. Mice were divided into four groups: bleomycin (BLM)-induced SSc (BLM-alone), BLM-induced SSc administered with mASCs (BLM-mASC), and BLM-induced SSc administered with mASCs activated with 10 or 100 μg/mL LMWH (BLM-hep-mASC); there were 9 mice per group ( n = 9). Skin inflammation and fibrosis were evaluated using histological and biochemical examinations and gene expression levels.
Results
In vitro assays showed that migration ability and HGF production were significantly higher in hep-mASCs than in mASCs alone. The mRNA expression levels of cell migration factors were significantly upregulated in hep-mASCs compared to those in mASCs alone. The hep-mASCs accumulated in the skin tissues more than mASCs alone. The thickness of skin and hydroxyproline content in BLM-hep-mASC groups were significantly decreased, and the skin mRNA expression levels of interleukin-2, α-smooth muscle actin, transforming growth factor β1, collagen type 1 alpha 1, and tissue inhibitor of metalloproteinase 2 were significantly downregulated compared to those in the BLM-alone group.
Conclusions
hep-mASCs showed higher anti-inflammatory and anti-fibrotic effects than mASCs alone and may be a promising candidate for SSc treatment.
... HGF exhibits anti-inflammatory effects by disrupting the NFkb pathway, decreasing IL-6 production, and increasing IL-10 production from CD14+ monocytes through the ERK1/2 pathway in various cells (35)(36)(37). In addition, HGF exhibits anti-fibrotic effects by inhibiting fibroblast proliferation and TGF-b production from fibroblasts (38,39). Therefore, in addition to the therapeutic effect of ASCs, HGF produced by hep-hASCs may have synergetic effects on LN by suppressing inflammation and fibrosis. ...
Background
Lupus nephritis is a life-threatening complication in systemic lupus erythematosus (SLE), but the efficiency of current therapies involving corticosteroids, immunosuppressants, and biological agents is limited. Adipose-derived mesenchymal stem cells (ASCs) are gaining attention as a novel treatment for inflammation in SLE. Low-molecular-weight heparin (LMWH) exhibits multiple functions including anti-inflammatory, anti-fibrotic, and cell function-promoting effects. LMWH stimulation is expected to increase the therapeutic effect of ASCs by promoting cellular functions. In this study, we investigated the effects of LMWH on ASC functions and the therapeutic effect of LMWH-activated human-ASCs (hep-hASCs) in an SLE mouse model.
Methods
The cellular functions of human-derived ASCs stimulated with different LMWH concentrations were observed, and the optimum LMWH dose was selected. The mice were assigned to control, human-ASC, and hep-hASC groups; treatments were performed on week 20. Twenty-six week-old mice were sacrificed, and urine protein score, serum blood urea nitrogen, creatinine (Cr), anti-ds DNA IgG antibody, and serum IL-6 levels were analyzed in each group. Mice kidneys were evaluated via histological examination, immunohistochemical staining, and gene expression levels.
Results
LMWH significantly promoted ASC migration and proliferation and hepatocyte growth factor production and upregulated immunomodulatory factors in vitro. Hep-hASC administration resulted in significant disease activity improvement including proteinuria, serum Cr and IL-6 levels, anti-ds DNA IgG antibody, glomerulonephritis, and immune complex in mice. Inflammation and fibrosis in kidneys was significantly suppressed in the hep-hASC group; the gene expression levels of TNF-alpha, TIMP-2, and MMP-2 was significantly downregulated in the hep-hASC group compared with the control group.
Conclusions
Hep-hASC exhibited higher anti-inflammatory and anti-fibrotic effects than hASCs and may be a candidate tool for SLE treatment in future.
... Activation of HGF resulted in phosphorylation of the HGF receptor (c-Met) 23 . Furthermore, the anti-fibrotic effect of the HGF/Met pathway is not restricted to the lung and has been consistently demonstrated in experimental models of liver, kidney, heart, and skin fibrosis [24][25][26][27][28][29] . The roles of HGF/ met signaling pathway have been reported. ...
Heparin and low molecular weight heparin (LMWH) have recently been considered useful treatment tools for inflammation. Heparin has antifibrotic activity, mediated by cellular secretion of hepatocyte growth factor (HGF). HGF has antifibrotic properties demonstrated in experimental models of lung, kidney, heart, skin, and liver fibrosis. The ability of LMWH for HGF secretion is similar to that of normal heparin. Poly (lactic-co-glycolic acid) (PLGA) is widely used for sustained drug release, because of its biocompatibility and low toxicity. LMWH-loaded PLGA microparticles are prepared by a conventional water-in-oil-in-water emulsion method. Interstitial pneumonia is a life-threatening pathological condition that causes respiratory failure when it progresses. In the present study, we investigated the therapeutic effect of LMWH-loaded PLGA microparticles in a mouse model of bleomycin-induced lung fibrosis. The ratios of fibrotic area to total area were significantly lower in mice administered LMWH-loaded microparticles than in mice administered bleomycin alone. The microparticle administration did not further enhance the gene expression for inflammatory cytokines. In a cell culture study, HGF secretion by mouse and human lung fibroblasts was significantly increased by LMWH addition. We conclude that LMWH showed anti-inflammatory activity, through the effects of LMWH-loaded PLGA microparticles on cells at sites of inflammation.
... This interaction subsequently turns on a variety of signaling pathways depending on cell types. HGF has been shown to play important roles in the regeneration process of various tissues by stimulating the proliferation and migration of respective progenitor cells (Kawaida et al., 1994;Huh et al., 2004;Wu et al., 2004;Watanabe et al., 2005). For example, HGF has been implicated in both skeletal muscle development and its regeneration after injury (Sisson et al., 2009). ...
Hepatocyte growth factor (HGF) is well known for its role in the migration of embryonic muscle progenitors and the activation of adult muscle stem cells, yet its functions during the adult muscle regeneration process remain to be elucidated. In this study, we showed that HGF/c-met signaling was activated during muscle regeneration, and that among various infiltrated cells, the macrophage is the major cell type affected by HGF. Pharmacological inhibition of the c-met receptor by PHA-665752 increased the expression levels of pro-inflammatory (M1) macrophage markers such as IL-1β and iNOS while lowering those of pro-regenerative (M2) macrophage markers like IL-10 and TGF-β, resulting in compromised muscle repair. In Raw 264.7 cells, HGF decreased the RNA level of LPS-induced TNF-α, IL-1β, and iNOS while enhancing that of IL-10. HGF was also shown to increase the phosphorylation of AMPKα through CaMKKβ, thereby overcoming the effects of the LPS-induced deactivation of AMPKα. Transfection with specific siRNA to AMPKα diminished the effects of HGF on the LPS-induced gene expressions of M1 and M2 markers. Exogenous delivery of HGF through intramuscular injection of the HGF-expressing plasmid vector promoted the transition to M2 macrophage and facilitated muscle regeneration. Taken together, our findings suggested that HGF/c-met might play an important role in the transition of the macrophage during muscle repair, indicating the potential use of HGF as a basis for developing therapeutics for muscle degenerative diseases.
... Among them, the skin tissue fibrosis and dermal thickening were the most important and common pathological features of SSc. Yamamoto and colleagues [43] used the Bleomycin (BLM) treatment to establish a new mouse model of SSc: subcutaneous injection of BLM can induce dermal and pulmonary fibrosis, autoantibody production and skin inflammatory infiltration, which was one of the best animal models for studying on systemic sclerosis. In this study, we used Bleomycin-induced mice on a C57BL/6 background to construct systemic sclerosis model. ...
Background
Systemic sclerosis (SSc) is an autoimmune disease characterized by fibrosis of the skin and internal organs. So far, no Western medicine treatment can completely inhibit or reverse the progress of SSc, while at the same time, our previous series of studies have shown that the treatment of SSc by the Wenyang Huazhuo Tongluo formula (WYHZTL), a Chinese herbal decoction, shows a delightful prospect. The aim of this study is to further investigate the mechanism of anti-fibrosis of WYHZTL formula in SSc mouse model.
Methods
The Bleomycin-induced SSc mouse model was treated with saline (BLM), high-dosage of WYHZTL formula (WYHZTL-H), medium-dosage of WYHZTL formula (WYHZTL-M), low-dosage of WYHZTL formula (WYHZTL-L) and XAV-939, a small molecule inhibitor of Wnt/β-catenin signaling pathway, by the intragastric administration and intraperitoneal injection, respectively. The mRNA and protein levels of Wnt/β-catenin signaling pathway associated genes, fibrosis markers and histopathology were detected by reverse transcription-quantitative polymerase chain reaction, Western blotting and hematoxylin/eosin-staining. The levels of Wnt1, CTGF and DKK1 protein in serum were detected by enzyme-linked immunosorbent assay.
Results
Compared with BLM group, the WYHZTL formula and XAV-939 could significantly inhibit the thickness of the skin tissue of the SSc mouse model. The mRNA expression levels of GSK3β and DKK1 in the WYHZTL formula and XAV-939-treated group were significantly higher than those in the BLM group, while Wnt1, β-catenin, TCF4, cyclin D1, survivin, VEGF, CTGF, FN1, collagen I/III were decreased. Compared with BLM group, the protein expression levels of GSK3β and DKK1 in the WYHZTL formula and XAV-939-treated group were upregulated, while Wnt1, β-catenin, cyclin D1, survivin, CTGF, FN1, collagen I/III were downregulated. WYHZTL formula and XAV-939 could inhibit expression of Wnt1 and CTGF, but promoted DKK1 in serum. Furthermore, WYHZTL-H seemed more effective than WYHZTL-M and/or XAV-939 on regulating Wnt1, β-catenin, TCF4, GSK3β, DKK1, cyclin D1, survivin, VEGF, CTGF, FN1 and collagen I/III.
Conclusion
This present study demonstrates that WYHZTL formula has anti-fibrosis effect in Bleomycin-induced SSc mouse model in a dosage-dependent manner, and the molecular mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.
Electronic supplementary material
The online version of this article (10.1186/s13020-018-0175-z) contains supplementary material, which is available to authorized users.
... Hepatocyte growth factor (HGF) is a multifunctional cytokine that is secreted by MSCs [13] and whose angiogenic, antiapoptotic, anti-fibrotic, and anti-inflammatory benefits have received increasing attention [14][15][16]. HGF acts as an anti-fibrotic agent that protects the host against transforming growth factor beta1 (TGF-b1)-mediated pro-fibrotic effects [14,17]. HGF plays an important role in the tissue-repair process, but the sustained delivery of HGF into the injured area remains a major challenge. ...
... Hepatocyte growth factor (HGF) is a multifunctional cytokine that is secreted by MSCs [13] and whose angiogenic, antiapoptotic, anti-fibrotic, and anti-inflammatory benefits have received increasing attention [14][15][16]. HGF acts as an anti-fibrotic agent that protects the host against transforming growth factor beta1 (TGF-b1)-mediated pro-fibrotic effects [14,17]. HGF plays an important role in the tissue-repair process, but the sustained delivery of HGF into the injured area remains a major challenge. ...
This study was designed to investigate the effects of hepatocyte growth factor (HGF) transgenic mesenchymal stem cells (HGF-MSCs) on wound healing in the sinonasal mucosa and nasal epithelial cells (NECs). We also sought to determine whether HGF-MSCs and MSCs can migrate into the injured mucosa and differentiate into ciliated cells. Human HGF-overexpressing umbilical cord MSCs (hHGF-UCMSCs) were established, and upregulation of hHGF expression was confirmed by RT-PCR and ELISA. To investigate the paracrine effect of human MSCs (hMSCs) on nasal epithelial repair, hMSC- and HGF-MSC-conditioned media (CM) were used in NEC proliferation assays and in an in vitro scratch-wound repair model. The in vivo sinonasal wound-healing model was established, and all enrolled rabbits were randomly assigned to four groups: the GFP-MSC group, the HGF-MSC group, the Ad-HGF group, and the surgery control group. The average decreased diameter was recorded, and the medial wall of the maxillary sinus was removed for histological analysis and scanning electron microscopy. Collagen deposition in the wound tissue was detected via Masson trichrome (M&T) staining. The distribution of MSCs and HGF-MSCs were observed by immunofluorescence. MSCs improved nasal wound healing both in vivo and in vitro. HGF overexpression in MSCs augmented the curative effects. Reduced collagen deposition and TGF-β1 expression were detected in the HGF-MSC group compared with the MSC-, Ad-HGF- and PBS-treated groups based on M&T staining and ELISA. HGF-MSCs and MSCs were capable of migrating to the injured tissues. HGF-MSCs promote wound healing both in vitro and in vivo. Moreover, MSCs inhibit collagen deposition in the maxillary sinus mucosa and are accompanied by decreased levels of the fibrogenic cytokine TGF-β1. In addition, both HGF-MSCs and MSCs can migrate to the injured mucosa and epithelial layer.
... The absence of IFNγ in the serum might suggest minimal systemic exposure to the protein; hence, side effects previously observed in the clinical trials could be avoided by topical administration of pDNA via the nanoparticulate delivery system. The present data suggest that topical application of IFNγ-coding pDNA using cationic gemini delivery vehicles significantly enhances cutaneous IFNγ levels after application to intact skin and appears to be comparable or more efficient than other non-viral delivery systems tested in vivo [87][88][89]. Additionally, its effect on collagen synthesis indicates the feasibility of this application to treat cutaneous involvement of scleroderma in humans. ...
Nonviral delivery systems must have higher transfection efficiencies than have been realized to date, in order to be useful in gene therapy. Gemini nanoparticles are composed of gemini surfactants and nucleic acids (DNAorRNA) with or without neutral lipid vesicles. This chapter provides potential nonviral delivery carriers for topical gene therapy applications for gemini nanoparticles. It discusses the physicochemical and transfection properties of the particle in detail. A section focuses on the application of cationic gemini nanoparticles as a gene delivery system for the treatment of localized scleroderma. Functionalization of gemini surfactants with fluorescent probes (e.g., pyrene) can also provide useful information on the intracellular fate of the nanoparticles, which aids in designing nanoparticles for subcellular intelligence.
... Indeed, it has been demonstrated, using different animal models of lung injury, that administration of exogenous HGF promotes lung repair and reduces fibrosis (Table 1) (62)(63)(64). In addition, the antifibrotic effect of the HGF/Met pathway is not restricted to the lung and has been consistently demonstrated in experimental models of liver (65), kidney (66), heart (67), and skin fibrosis (68,69). ...
Idiopathic pulmonary fibrosis is currently believed to be driven by alveolar epithelial cells, with abnormally activated alveolar epithelial cells accumulating in an attempt to repair injured alveolar epithelium (1). Thus, targeting the alveolar epithelium to prevent or inhibit the development of pulmonary fibrosis might be an interesting therapeutic option in this disease. Hepatocyte growth factor (HGF) is a growth factor for epithelial and endothelial cells, which is secreted by different cell types, especially fibroblasts and neutrophils. HGF has mitogenic, motogenic, and morphogenic properties and exerts an antiapoptotic action on epithelial and endothelial cells. HGF has also proangiogenic effect. In vitro, HGF inhibits epithelial-to-mesenchymal cell transition and promotes myofibroblast apoptosis. In vivo, HGF has antifibrotic properties demonstrated in experimental models of lung, kidney, heart, skin, and liver fibrosis. Hence, the modulation of HGF may be an attractive target for the treatment of lung fibrosis.
... Along with a series of in vitro experiments using cultured cells from different organs, various animal models (induced/ injured and spontaneous/uninjured) of tissue fibrosis have been used to explore the molecular basis of fibrosis in vivo (Ellmers, 2010;Leask, 2010;Mastuzaki, 2010;Yang et al., 2010;Yue et al., 2010). The widely used animal models of tissue fibrosis include the following: For skin and lung fibrosis, the bleomycin-induced mouse model (Murray et al., 2008;Wu et al., 2004aWu et al., ,2009Yamamoto et al., 1999) and the graft-versus-host disease (GVHD) mouse model (McCormick et al., 1999); for cardiac fibrosis, the transverse aortic banding (TAB) model (Sun et al., 2007), angiotensin II (AngII)-induced model , phenylephrine-induced model (Farivar et al., 1995), isoproterenol-induced model (Rohrer et al., 1996), and aldosterone-induced model ; for kidney fibrosis, the unilateral ureteral obstruction (UUO)-induced model and aldosterone-induced renal injury model (Ma et al., 2006;Ma and Fogo, 2009;Oda et al., 2001); for liver fibrosis, the carbon tetrachloride (CCl 4 )-induced mouse model and bile duct ligation model (Wang et al., 2007); and for gastrointestinal tract fibrosis, the chronic 2,4,6-trinitrobenzene sulfonic acid-induced colitis model (Wu and Chakravarti, 2007). Besides the use of induced models of tissue fibrosis, spontaneously developed tissue fibrosis has also been reported in genetically modified animals. ...
Fibrosis is defined as a fibroproliferative or abnormal fibroblast activation-related disease. Deregulation of wound healing leads to hyperactivation of fibroblasts and excessive accumulation of extracellular matrix (ECM) proteins in the wound area, the pathological manifestation of fibrosis. The accumulation of excessive levels of collagen in the ECM depends on two factors: an increased rate of collagen synthesis and or decreased rate of collagen degradation by cellular proteolytic activities. The urokinase/tissue type plasminogen activator (uPA/tPA) and plasmin play significant roles in the cellular proteolytic degradation of ECM proteins and the maintenance of tissue homeostasis. The activities of uPA/tPA/plasmin and plasmin-dependent MMPs rely mostly on the activity of a potent inhibitor of uPA/tPA, plasminogen activator inhibitor-1 (PAI-1). Under normal physiologic conditions, PAI-1 controls the activities of uPA/tPA/plasmin/MMP proteolytic activities and thus maintains the tissue homeostasis. During wound healing, elevated levels of PAI-1 inhibit uPA/tPA/plasmin and plasmin-dependent MMP activities, and, thus, help expedite wound healing. In contrast to this scenario, under pathologic conditions, excessive PAI-1 contributes to excessive accumulation of collagen and other ECM protein in the wound area, and thus preserves scarring. While the level of PAI-1 is significantly elevated in fibrotic tissues, lack of PAI-1 protects different organs from fibrosis in response to injury-related profibrotic signals. Thus, PAI-1 is implicated in the pathology of fibrosis in different organs including the heart, lung, kidney, liver, and skin. Paradoxically, PAI-1 deficiency promotes spontaneous cardiac-selective fibrosis. In this review, we discuss the significance of PAI-1 in the pathogenesis of fibrosis in multiple organs.
... Growing evidence shows that HGF elicits the regression of fibrosis in numerous organs, such as scleroderma,105) cardiomyopathy,74) vocal scarring and peritoneal fibrosis, in which TGF-β-induced MyoFBs are critical for tissue scarring. The resolution of fibrosis provides an open space for repair of the epithelium and endothelium, resulting in recovery from organ failure.81) ...
It has been more than 25 years since HGF was discovered as a mitogen of hepatocytes. HGF is produced by stromal cells, and stimulates epithelial cell proliferation, motility, morphogenesis and angiogenesis in various organs via tyrosine phosphorylation of its receptor, c-Met. In fetal stages, HGF-neutralization, or c-Met gene destruction, leads to hypoplasia of many organs, indicating that HGF signals are essential for organ development. Endogenous HGF is required for self-repair of injured livers, kidneys, lungs and so on. In addition, HGF exerts protective effects on epithelial and non-epithelial organs (including the heart and brain) via anti-apoptotic and anti-inflammatory signals. During organ diseases, plasma HGF levels significantly increased, while anti-HGF antibody infusion accelerated tissue destruction in rodents. Thus, endogenous HGF is required for minimization of diseases, while insufficient production of HGF leads to organ failure. This is the reason why HGF supplementation produces therapeutic outcomes under pathological conditions. Moreover, emerging studies delineated key roles of HGF during tumor metastasis, while HGF-antagonism leads to anti-tumor outcomes. Taken together, HGF-based molecules, including HGF-variants, HGF-fragments and c-Met-binders are available as regenerative or anti-tumor drugs. Molecular analysis of the HGF-c-Met system could provide bridges between basic biology and clinical medicine.
(Communicated by Kunihiko SUZUKI, M.J.A.)
