Hepatocyte Growth Factor Group Research Articles

Chronic treatment with a combination of hepatocyte growth factor and JNK inhibitor ameliorates cavernosal veno-occlusive dysfunction: a rat model of cavernous nerve injury.

Structural alterations of the penis, including cavernosal apoptosis and fibrosis, induce venous leakage into the corpus cavernosum or cavernosal veno-occlusive dysfunction, a key pathophysiology associated with erectile dysfunction after radical prostatectomy. We hypothesized that the effect of JNK inhibitors on reducing apoptosis and hepatocyte growth factor (HGF) on inducing tissue regeneration could be another treatment mechanism of erectile dysfunction after radical prostatectomy. To investigate whether JNK inhibition combined with intracavernosal administration of HGF can completely preserve cavernosal veno-occlusive function (CVOF) in a rat model of erectile dysfunction induced via bilateral cavernosal nerve crush injury (CNCI). A total of 42 male Sprague-Dawley rats were randomly assigned to sham control (group S), CNCI (group I), and CNCI treated with a combination of JNK inhibitor and HGF (group J + H) for 5weeks after surgery. Rats in each group were evaluated via dynamic infusion cavernosometry (DIC), caspase-3 activity assay, Masson trichrome staining, immunohistochemical staining of α-smooth muscle actin, and immunoblotting at 5weeks after surgery. Regarding CVOF, group I showed decreased papaverine response, increased maintenance, and drop rates of DIC when compared with group S. Group J + H showed significant improvement in the 3 DIC parameters vs group I. No differences in the 3 DIC parameters were found between group J + H and group S. Regarding the structural integrity of the corpus cavernosum, group I showed increased caspase-3 activity, decreased smooth muscle (SM):collagen ratio, decreased SM content, decreased protein expression of PECAM-1, and decreased phosphorylation of c-Jun and c-Met. Group J + H showed significant attenuation in histologic and molecular derangement as compared with group I. There were no differences in caspase-3 activity, SM content, SM:collagen ratio, PECAM-1 protein expression, c-Jun phosphorylation, and c-Met phosphorylation between groups J + H and S. Our results suggest that antiapoptotic and regenerative therapy for the corpus cavernosum is a potential mechanism of penile rehabilitation after radical prostatectomy. This study provides evidence that combination treatment of JNK inhibitor and HGF preserves erectile function by restoring corporal SM and endothelium. However, additional human studies are needed to confirm the clinical effect. Chronic treatment with JNK inhibitor and HGF may preserve CVOF to levels comparable to sham control by preserving the structural integrity of the corpus cavernosum and so represents a potential therapeutic option for preventing the development of cavernosal veno-occlusive dysfunction.

Abstract 13947: Heparin-Mediated Release of Hepatocyte Growth Factor at the Acute Phase of STEMI

Introduction: Hepatocyte Growth Factor (HGF) is a cytokine promoting cell survival via its receptor c-met. Heparin injection is known to induce a massive release of native HGF in blood. Heparin is used at the acute phase of myocardial infarction (MI). However, HGF release at the acute phase of MI and its potential cardioprotective effect is unknown. Hypothesis: We aimed to assess HGF kinetics in a cohort of ST-elevation myocardial infarction (STEMI) and its potential cardioprotective effect. Methods: We prospectively included consecutive STEMI patients admitted in our hospital from 2016 to 2019. All patients underwent coronary angiography with revascularization. Sera were collected at 5 time points (admission, 4 hours (H4), H24, H48 and 1 month after STEMI). Samples were stored at -80°C. HGF levels were assess by ELISA. We used a mouse model of myocardial ischemia reperfusion (ligation of the left anterior descending coronary artery followed by reperfusion) to evaluate HGF effect. Results: We included 230 patients. We observed an intense and early peak of HGF as early as admission, before revascularization (8750 pg/ml, IQR [8021-9492]) followed by a rapid decrease within the first 48h (p<0.0001). We demonstrated that this rapid increase was induced by heparin administration (unfractionated heparin or low molecular weight heparins). To evaluate the effect of HGF on myocardial infarct size we administrated HGF or placebo in mice during myocardial reperfusion. In the HGF group, infarct size was significantly smaller compared to the control group (52.0% of left ventricle vs 58.0%, p=0.002). Conclusions: We demonstrated an intense and early HGF peak at the acute phase of STEMI, induced by heparin. In our preclinical mouse model, HGF had cardioprotective effect after MI. Thus we may suppose that heparin-mediated release of HGF at the acute phase of STEMI have cardioprotective effect that may confer a basal cardioprotection in all patients.

P-023 Exogenous glial cell-derived neurotrophic factor contributes hepatocyte growth factor-induced in vitro organization of Sertoli cells on matrigel-based three-dimensional culture

Abstract Study question Does glial cell-derived neurotrophic factor (GDNF), a crucial factor for spermatogonial stem cell (SSC) self-renewal, have a role in in vitro tubule-formation capacity of Sertoli cells? Summary answer Exogenous GDNF treatment together with hepatocyte growth factor (HGF) significantly increased the in vitro tubule-forming capacity of Sertoli cells on matrigel-based three-dimensional (3D) culture system. What is known already GDNF increases the tubule-forming capacity of HGF in vitro by regulating c-Met expression during renal tubulogenesis. In vitro testicular organization of Sertoli cells consists of two stages: testicular cord-like structures and tubule-like structures. HGF secreted from Sertoli cells plays a direct role in forming seminiferous tubules in vivo and in vitro by activating c-Met. GDNF maintains SSC numbers by activating GFRα1 and Ret receptors in SSCs and can also directly phosphorylate c-Met in GFRα1-positive cells. Study design, size, duration Four groups were established using the 15P-1 Sertoli cell line: Control, HGF-administrated, GDNF-administrated and, HGF and GDNF-administrated. All groups were pursued on a matrigel-based 3D culture system for seven days. Experiments were repeated three times in all groups. Kit ligand (KitL, Sertoli cell marker; positive control) and c-kit (germ cell marker; negative control) were used as the control for Sertoli cell characterization. This work was supported by TUBITAK (SBAG-120S554) and Akdeniz University Research Foundation (TYL-2020-5481). Participants/materials, setting, methods The cord-forming and tubule-forming capacity of Sertoli cells on matrigel-based 3D culture was evaluated under an inverted microscope, and c-Met and p-c-Met expression levels were analyzed using western blot. In addition to KitL and c-kit protein expression, zonula occludens-1 (ZO-1) protein expression for polarization of Sertoli cells was evaluated by immunofluorescence staining. Data were analyzed using one-way ANOVA and Tukey post hoc test for multiple comparisons. p < 0.05 was considered statistically significant. Main results and the role of chance In the first 24 hours, interaction and aggregation between Sertoli cells began. The highest cellular interaction was found in the group treated with HGF and GDNF compared to the other groups (p < 0.05). At 48 and 72 hours, the formation of cord-like structures was observed by forming connections between the clusters formed in all groups in the first 24 hours. The number of cord-like structures was higher in the groups treated with only HGF and HGF and GDNF at 48th and 72nd hours (p < 0.05). At 96th and 120th hours, structures forming spaces were observed, and these structures were determined to be lumen-like structures. The capacity of 15P-1 Sertoli cells to form tubule-like structures was higher in the HGF and GDNF treated groups than in the other groups (p < 0.05). c-Met and p-c-Met protein expression levels were high in HGF, GDNF, HGF, and GDNF groups compared to control (P < 0.05). Sertoli cells retained their identity until the end of the culture and formed tight junctions in the HGF and GDNF-administrated group as evaluated by ZO-1 expression. Limitations, reasons for caution The cord- and tubule-forming capacity of Sertoli cells was evaluated in vitro utilizing exogenous GDNF treatment but the effect of testicular endothelial cell-derived GDNF should be also evaluated. Wider implications of the findings Exogenous GDNF treatment enhances HGF-induced in vitro tubule-formation capacity of Sertoli cells by directly phosphorylating c-Met in Sertoli cells. Our findings highlight the importance of designing studies related to GDNF-induced cell signaling pathway for in vitro tubulogenesis of Sertoli cells to support ex vivo spermatogenesis. Trial registration number None

Abstract 5138: Cetuximab resistance induced by hepatocyte growth factor is overcome by MET inhibition in KRAS, NRAS, and BRAF wild-type colorectal cancer

Abstract Background: Recent evidence has highlighted the role of hepatocyte growth factor (HGF) as a putative biomarker to predict EGFR inhibitor resistance. This study investigated the impact of plasma HGF levels on EGFR inhibition and the therapeutic implications of MET inhibition in KRAS, NRAS, and BRAF (RAS/RAF) wild-type colorectal cancer (CRC). Methods: Clinical outcomes were analyzed according to the plasma HGF levels in patients with metastatic CRC (mCRC) receiving palliative first-line cetuximab or bevacizumab + FOLFIRI chemotherapy. Then, in vitro experiments were conducted to validate the clinical findings and to establish pre-clinical evidence of MET inhibition in the HGF-high population. Results: From March 2015 to September 2019, a total of 80 patients were enrolled. The median age was 63 (range, 24-85) years. Of these, 45 patients (56.2%) were treated with bevacizumab + FOLFIRI and 35 patients (43.8%) were treated with cetuximab + FOLFIRI. Among patients with evaluable disease (43 patients (95.6%) in the bevacizumab group and 34 patients (97.1%) in the cetuximab group), the objective response rate was 60.0% and 37.8%, respectively (p=0.162). The median serum HGF level was 374.75 (range, 9.43-30,644.44) pg/mL, and the optimal cut-off value of HGF levels was 12.70 pg/mL by the maximal chi-square method. During a median follow-up of 29.3 (range, 1.2-71.3) months, both progression-free survival (PFS) and overall survival (OS) were significantly shorter in the high HGF group compared with the low HGF group: median 11.8 (95% confidence interval [CI], 10.5-13.1) months vs. 24.7 (95% CI, 23.4-25.9) months for PFS (p=0.009), and median 21.1 (95% CI, 17.9-24.3) months vs. not reached for OS (p=0.018). The difference in PFS and OS was statistically significant in the cetuximab group (p=0.003 for PFS and p=0.035 for OS), but not in the bevacizumab group. In five RAS/RAF wild-type CRC cell lines (Caco-2, COLO 320DM, KM12C, SNU-C1, and SNU-C4), the addition of HGF activated ERK1/2 and AKT via MET phosphorylation. In the cytotoxicity assays, Caco-2 and SNU-C4 were relatively sensitive to cetuximab compared with the others and, in the presence of HGF, cetuximab sensitivity was significantly decreased in the two cells. Moreover, capmatinib, a MET inhibitor, abrogated the effect of HGF on common downstream signaling pathways of EGFR and MET and thus the cetuximab resistance was significantly overcome in vitro. Conclusions: Among patients with mCRC receiving cetuximab + FOLFIRI, those with high plasma HGF levels had significantly worse PFS and OS. HGF induced cetuximab resistance by AKT and ERK activation while capmatinib, a MET inhibitor, significantly increased the anti-tumor effects of cetuximab in the presence of HGF in vitro. Our data may provide evidence of future clinical trials of dual EGFR and MET targeting strategies in patients with HGF-high, RAS/RAF wild-type mCRC. Citation Format: Sang-A Kim, Hyejoo Park, Kui-Jin Kim, Ji-Won Kim, Ji Hea Sung, Milang Nam, Ju Hyun Lee, Eun Hee Jung, Koung Jin Suh, Ji Yun Lee, Se Hyun Kim, Jeong-Ok Lee, Jin Won Kim, Yu Jung Kim, Jee Hyun Kim, Soo-Mee Bang, Jong Seok Lee, Keun-Wook Lee. Cetuximab resistance induced by hepatocyte growth factor is overcome by MET inhibition in KRAS, NRAS, and BRAF wild-type colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5138.

MP38-03 COMBINATION TREATMENT WITH A JNK INHIBITOR AND HEPATOCYTE GROWTH FACTOR FOR RESTORATION OF ERECTILE FUNCTION IN A RAT MODEL OF CAVERNOSAL NERVE INJURY: COMPARISON WITH A JNK INHIBITOR ALONE OR HEPATOCYTE GROWTH FACTOR ALONE

You have accessJournal of UrologyCME1 May 2022MP38-03 COMBINATION TREATMENT WITH A JNK INHIBITOR AND HEPATOCYTE GROWTH FACTOR FOR RESTORATION OF ERECTILE FUNCTION IN A RAT MODEL OF CAVERNOSAL NERVE INJURY: COMPARISON WITH A JNK INHIBITOR ALONE OR HEPATOCYTE GROWTH FACTOR ALONE Junghoon Lee, Sangjun Yoo, Min Soo Choo, Hwancheol Son, Hyeon Jeong, Soo Woong Kim, and Min Chul Cho Junghoon LeeJunghoon Lee More articles by this author , Sangjun YooSangjun Yoo More articles by this author , Min Soo ChooMin Soo Choo More articles by this author , Hwancheol SonHwancheol Son More articles by this author , Hyeon JeongHyeon Jeong More articles by this author , Soo Woong KimSoo Woong Kim More articles by this author , and Min Chul ChoMin Chul Cho More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002592.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The structural derangements of the corpus cavernosum are known to be potential causes of intractable impotence or to limit efficacy of PDE5 inhibitors by inducing cavernosal venoocclusive dysfunction, a key pathophysiology of postprostatectomy erectile dysfunction. The aim of this study was to determine if combined administration of a JNK inhibitor and HGF (hepatocyte growth factor) would restore erectile function through both antiapoptotic and regenerative effects in a rat model of erectile dysfunction induced by cavernous nerve crush injury (CNCI), and to compare it with administration of a JNK inhibitor alone or HGF alone. METHODS: We randomized 70 rats into five experimental groups for two weeks from the day of CNCI: a sham surgery group (S), a CNCI (I) group, a group treated with once-daily intraperitoneal administration of 10.0 mg/kg of JNK inhibitor (J), a twice-weekly intracavernosal administration of 4.2 μg HGF group (H), and a combined treatment with 10.0 mg/kg JNK inhibitor and 4.2 μg HGF group (J+H). We investigated erectile responses to electrostimulation, histological staining, caspase-3 activity assay, and immunoblotting at two weeks following surgery. RESULTS: The three treatment groups showed improvements in the erectile responses (ICP/MAP or AUC/MAP ratio) after stimulation at all voltages as compared to the I group. The erectile responses in the J+H group were greater than those in the J or H group. The erectile responses in the J+H group were not generally different from those in the S-group. Regarding the cavernosal apoptosis, the caspase-3 activity and c-Jun phosphorylation in the J and J+H groups were improved as compared to the I group, whereas the caspase-3 activity in the H group partially improved. Regarding the endothelial status and smooth muscle (SM) content, the protein expression of PECAM-1, eNOS phosphorylation, and α-SM actin content in the J+H group were normalized, although those in the J or H group were partially restored. Phosphorylation of cMet was normalized in the H and J+H groups. CONCLUSIONS: Our data indicates that combination therapy with a JNK inhibitor and HGF can generally normalize erectile function through antiapoptosis and the preservation of cavernosal endothelium or SM in a rat CN injury model. The combined treatment with these two kinds of agents also appears to be superior to the respective agent alone in terms of the therapeutic effects. Source of Funding: This research was supported by the National Research Foundation of Korea, grant 2018R1D1A1A09083833, and by the JEIL Pharmaceutical Co., LTD, grant 01-2018-33 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e619 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Junghoon Lee More articles by this author Sangjun Yoo More articles by this author Min Soo Choo More articles by this author Hwancheol Son More articles by this author Hyeon Jeong More articles by this author Soo Woong Kim More articles by this author Min Chul Cho More articles by this author Expand All Advertisement PDF DownloadLoading ...

The preventive effect of recombinant human hepatocyte growth factor for hepatic steatosis in a rat model of short bowel syndrome

PurposeShort bowel syndrome (SBS) patients require total parenteral nutrition (TPN) following massive small bowel resection (SBR), which may cause intestinal failure-associated liver disease (IFALD), a life-threatening complication. Hepatocyte growth factor (HGF) acts as a potent hepatocyte mitogen with anti inflammatory and antioxidant actions. The present study evaluated the effect of recombinant human HGF (rh-HGF) on SBR and subsequent IFALD using a parentally fed rat model of SBS. MethodsRats underwent jugular vein catheterization for continuous TPN and 90% SBR. They were divided into 2 groups: TPN alone (SBS/TPN group: n = 7) or TPN plus the intravenous administration of rh-HGF (0.3 mg/kg/day) (SBS/TPN+HGF group: n = 7). On day 7, their tissues and stool were harvested to evaluate the effects of HGF. ResultsRegarding the histological findings, based on the nonalcoholic fatty liver disease (NAFLD) activity score, the SBS/TPN+HGF group showed significantly less hepatic steatosis and inflammatory cell infiltration than the SBS/TPN group (NAFLD activity score, 4.00 ± 1.83 vs. 1.00 ± 0.82; p < 0.01). The SBS/TPN+HGF group showed a higher expression of Farnesoid X receptor in the liver and lower expression of Toll-like receptor 4 in the ileum than the SBS/TPN group. Regarding the composition of the bacterial gut microbiota, Actinobacteria, Bacteroidetes and Proteobacteria were decreased in the SBS/TPN+HGF group compared with the SBS/TPN group. ConclusionIn our SBS with TPN rat model, rh-HGF administration had a preventive effect against hepatic steatosis and dysbiosis. rh-HGF may therefore be a potentially effective therapeutic agent for SBS and subsequent IFALD. Type of studyExperimental research.

Cetuximab resistance induced by hepatocyte growth factor is overcome by MET inhibition in KRAS, NRAS, and BRAF wild-type colorectal cancers.

Recent evidence has highlighted the role of hepatocyte growth factor (HGF) as a putative biomarker to predict EGFR inhibitor resistance. This study investigated the impact of plasma HGF levels on EGFR inhibition and the counter effect of MET inhibition in KRAS, NRAS, and BRAF (RAS/RAF) wild-type colorectal cancers (CRCs). Plasma HGF levels were analyzed with clinical outcomes of patients with metastatic CRC (mCRC) receiving palliative first-line chemotherapy. Then, in vitro experiments were conducted to validate the clinical findings and to establish pre-clinical evidence of MET inhibition by capmatinib. A total of 80 patients were included: cetuximab + FOLFIRI (n = 35) and bevacizumab + FOLFIRI (n = 45). Both progression-free survival (PFS) and overall survival (OS) were significantly lesser in the high vs low HGF group: median 11.8 vs. 24.7months, respectively, for PFS (p = 0.009), and median 21.1months vs. not reached, respectively, for OS (p = 0.018). The difference was significantly evident in the cetuximab group. In five RAS/RAF wild-type CRC cells, the addition of HGF activated ERK1/2 and AKT via MET phosphorylation, resulting in cetuximab resistance in vitro. In cetuximab-sensitive Caco-2 and SNU-C4 cells, capmatinib overcame cetuximab resistance in the presence of HGF by attenuating HGF-induced MET signaling activation. Patients with mCRC receiving cetuximab + FOLFIRI who presented with high plasma HGF levels had significantly worse PFS and OS. Cetuximab resistance induced by HGF was mediated by AKT and ERK activation and overcome by MET inhibition in vitro.

Combination Therapy with a JNK Inhibitor and Hepatocyte Growth Factor for Restoration of Erectile Function in a Rat Model of Cavernosal Nerve Injury: Comparison with a JNK Inhibitor Alone or Hepatocyte Growth Factor Alone

We determined if combined administration of JNK-inhibitors and HGF (hepatocyte-growth-factor) would restore erectile-function through both antiapoptotic and regenerative effects in a rat model of cavernous-nerve-crush-injury (CNCI), and compared the results with administration of JNK-inhibitor alone or HGF alone. We randomized 70 rats into 5 groups: sham-surgery-group (S), CNCI (I) group, a group treated with once-daily intraperitoneal-administration of 10.0-mg/kg of JNK-inhibitors (J), a twice-weekly intracavernosal-administration of 4.2-μg HGF group (H), and a combined-treatment with 10.0-mg/kg JNK-inhibitors and 4.2-μg HGF group (J+H). We investigated erectile-responses to electrostimulation, histological-staining, caspase-3-activity-assay, and immunoblotting at two-weeks postoperatively. The three treatment groups showed improvements in erectile-responses (ICP/MAP and AUC/MAP ratios) compared to Group-I. The erectile-responses in Group-J+H were greater than those in Group-J or Group-H. The erectile-responses in Group-J+H were generally normalized. Caspase-3-activity and cJun-phosphorylation in Group-J and Group-J+H improved compared to Group-I, whereas caspase-3-activity in Group-H partially improved. Protein-expression of PECAM-1, eNOS-phosphorylation, and smooth-muscle content in Group-J+H were normalized, although those in Group-J or Group-H were partially restored. Combination therapy with JNK-inhibitors and HGF can generally normalize erectile-function through anti-apoptosis and preservation of endothelium or SM in rat CNCI model. The combined treatment appears to be superior to the respective agent alone in terms of therapeutic effects.

Hepatocyte growth factor and soluble cMet levels in plasma are prognostic biomarkers of mortality in patients with severe acute kidney injury.

BackgroundHepatocyte growth factor (HGF)/cMet pathway is necessary for repair and regeneration following acute kidney injury (AKI). We evaluated the clinical potential of plasma HGF and soluble cMet as prognostic biomarkers for severe AKI requiring continuous renal replacement therapy (CRRT).MethodsOne hundred thirty-six patients with severe AKI who participated in the VENUS (volume management under body composition monitoring in critically ill patients on CRRT) trial between 2017 and 2019 were enrolled in this study. We investigated associations between plasma HGF and cMet concentrations and all-cause mortality.ResultsPlasma HGF and soluble cMet levels were positively correlated. Patients were divided into three groups based on their HGF and soluble cMet concentrations. The day D 0, D2, and D7 highest concentration HGF groups had significantly higher in-hospital mortality after adjusting for sex, body mass index, Acute Physiology and Chronic Health Evaluation II, and age-adjusted Charlson comorbidity index score, especially on D7 (hazard ratio, 4.26; 95% confidence interval, 1.71–10.62; p = 0.002). D7 soluble cMet level was also associated with mortality. Receiver operating characteristic curve analysis indicated that D7 HGF and soluble cMet levels were best at predicting mortality. Addition of plasma HGF and soluble cMet to conventional prognostic indices significantly improved the predictive value for mortality on D7. However, plasma HGF and soluble cMet were not associated with fluid status.ConclusionPlasma HGF and soluble cMet levels were significant predictors of the outcomes of severe AKI patients undergoing CRRT. There was no correlation between plasma HGF and soluble cMet levels and fluid balance.

Serum hepatocyte growth factor as a predictor of disease severity and future exacerbations in patients with non-cystic fibrosis bronchiectasis

BackgroundSerum biomarkers associated with the severity of non-cystic fibrosis (CF) bronchiectasis are insufficient. This study determined the association of serum hepatocyte growth factor (HGF), osteopontin, and pentraxin-3 levels with disease severity and exacerbation in patients with non-CF bronchiectasis. MethodsSerum levels of HGF, osteopontin, and pentraxin-3 were measured in patients with clinically stable non-CF bronchiectasis (n = 61). The correlation between the biomarkers and bronchiectasis severity index (BSI) and FACED score was assessed using univariate and multivariate linear regression analyses. Predictive variables associated with exacerbation were analyzed using a Cox proportional hazards model and the time to first exacerbation in high and low HGF groups during the observation period was compared using Kaplan-Meier survival curves. ResultsThe BSI showed significant correlation with HGF (r = 0.423; p = 0.001) and pentraxin-3 (r = 0.316; p = 0.013). The FACED score was significantly correlated with HGF (r = 0.406; p = 0.001). Univariate and multivariate linear regression analysis revealed that serum level of HGF was independently associated with both scoring systems. The high HGF group showed a significantly shorter time to first exacerbation (Log-rank test, p = 0.014). Multivariate Cox proportional hazards regression analysis revealed that high serum HGF level and colonization with non-pseudomonas organisms were independent predictors of future exacerbations (HR 2.364; p = 0.024 and HR 2.438; p = 0.020, respectively). ConclusionSerum level of HGF is a potential biomarker that is closely associated with disease severity and future risk of exacerbations in patients with non-CF bronchiectasis.

The administration of hepatocyte growth factor prevents total parenteral nutrition-induced hepatocellular injury in a rat model

Total parenteral nutrition (TPN) sometimes induces parenteral nutrition-associated liver disease (PNALD). Hepatocyte growth factor (HGF) acts as a potent hepatocyte mitogen anti-inflammatory and antioxidant actions. We aimed to evaluate the effect of HGF on PNALD in a rat model of TPN. A catheter was placed in the right jugular vein for 7-day continuous TPN. All rats were divided into three groups: TPN alone (TPN group), TPN plus intravenous HGF at 0.3mg/kg/day [TPN + HGF (low) group], and TPN plus HGF at 1.0mg/kg/day [TPN + HGF (high) group]. On day 7, livers were harvested and the histology, inflammatory cytokines and apoptosis were evaluated. Histologically, lipid droplets were apparent in the TPN group, but decreased in the TPN + HGF (low) and TPN + HGF (high) groups. The histological nonalcoholic fatty liver disease activity scores in the TPN + HGF (low) and TPN + HGF (high) groups were significantly lower than that in the TPN group (p < 0.01). There were no significant differences in the inflammatory cytokine levels of the three groups. The caspase-9 expression levels in the TPN + HGF (low) and TPN + HGF (high) groups were significantly decreased in comparison to that in the control group (p < 0.05). The intravenous administration of HGF attenuated hepatic steatosis induced by 7-day TPN dose dependently.

Effect of Human Umbilical Cord Mesenchymal Stem Cells Transfected with HGF on TGF-β1/Smad Signaling Pathway in Carbon Tetrachloride-Induced Liver Fibrosis Rats.

The research on human umbilical cord-derived mesenchymal stem cells (hUCMSCs) suggests promising therapeutic strategy for ameliorating liver fibrosis and it can be an effective alternative method of orthotopic liver transplantation. Hepatocyte growth factor (HGF) is the most basic cytokine involved in the inhibition of liver fibrosis and promotion of hepatocyte proliferation and regeneration. The objective of this study was to determine the possible mechanism about how the microencapsulated hUCMSCs made by alginate-poly-lysine-alginate (A-P-A) transfected with HGF could ameliorate liver fibrosis through the TGF-β1/Smad signaling pathway. The microencapsulated cells were divided into four groups: hUCMSC (microcapsules of hUCMSCs), HGF (microcapsules of HGF+hUCMSCs), LV5-NC (microcapsules of LV5-NC, an rLV-EF1a-EGFP+Puro control lentiviral vector+hUCMSCs), and empty microcapsule (microcapsules without any hUCMSCs), and then transplanted by intraperitoneal injection into carbon tetrachloride (CCl4)-induced liver fibrosis rats, respectively. The results showed that the fibrosis in the hUCMSC, LV5-NC, and HGF groups was significantly alleviated. Moreover, the messenger RNA (mRNA) and protein levels of collagen I, collagen III, α-SMA, TGF-β1, Smad2, and Smad3 were significantly decreased compared with the empty microcapsule group and these indices in HGF group were more decreased compared with hUCMSC and LV5-NC groups. This study indicated that microencapsulated hUCMSCs transfected with HGF could effectively improve CCl4-induced rat liver fibrosis and the possible mechanism was closely related to the inhibition of TGF-β1/Smad signaling pathway.

Hepatocyte growth factor predicts failure of Fontan circulation.

AimsThis study aimed to assess the value of the hepatocyte growth factor (HGF) as an independent predictor of a Fontan circulation failure.Methods and resultsThis retrospective case–control study included 34 consecutive patients (19 men and 15 women) who underwent a post‐operative cardiac catheterization after a Fontan operation at the Saitama Medical University International Medical Center between April 2017 and December 2019. We divided the patients into two groups according to the HGF level: HGF < 0.4 ng/mL (n = 20, normal HGF group) and HGF ≥ 0.4 ng/mL (n = 14, elevated HGF group). The age at the time of the cardiac catheterization was 59.3 ± 7.9 months. The range of the duration between the Fontan operation and the cardiac catheterization was 37.5 ± 7.9 months. The age (P = 0.417), gender (P = 0.08), morphology of the functional ventricle (P = 0.99), presence or closure of the Fontan fenestration (P = 0.704), and rate of medication use (angiotensin‐converting enzyme inhibitors or angiotensin II receptor blockers) (P = 0.99) were equivalent between the two groups. Laboratory parameters including the brain natriuretic peptide level (P = 0.085), serum creatinine level (P = 0.27), and aspartate aminotransferase level (P = 0.235) were similar between the two groups. The elevated HGF group had a higher C‐reactive protein level than the normal HGF group (0.42 ± 0.14 and 0.05 ± 0.01 mg/dL, P = 0.005). The elevated HGF group had a higher central venous pressure (CVP) level than the normal HGF group (13.4 ± 0.7 and 9.7 ± 0.4 mmHg, P < 0.0001), and the HGF was positively correlated with the CVP (P = 0.0004, r 2 = 0.33). The SvO2 level was significantly lower in the elevated HGF group than in the normal HGF group (61.9 ± 2.3% and 75.0 ± 1.2%, P < 0.0001), and the HGF was negatively correlated with the SvO2 (P < 0.0001, r 2 = 0.65). Of the 34 patients, six underwent catheter interventions. Patients who underwent catheter interventions had a higher HGF level than those who did not (0.44 ± 0.03 and 0.37 ± 0.01 ng/mL, P = 0.032). The receiver operating characteristic curve created for the discrimination of a catheter intervention revealed that an HGF value of >0.405 ng/mL could detect the need for a catheter intervention with 75.0% sensitivity and 83.3% specificity. A multivariable regression analysis showed that an elevated HGF was an independent predictor of an elevated CVP (β‐coefficient 21.2, SE 5.5, P = 0.0005) and decreased SvO2 (β‐coefficient −92.9, SE 12.4, P < 0.0001).ConclusionsThe HGF is an independent predictor of a failure of a Fontan circulation. The HGF is an indicator for an additional catheter intervention after a Fontan operation.

The Efficacy of Topical HGF on Corneal Fibrosis and Epithelial Healing after Scar-Producing PRK Injury in Rabbits.

PurposeTo determine the effect of topical hepatocyte growth factor (HGF) on myofibroblast development and corneal opacity after fibrosis-producing photorefractive keratectomy (PRK).MethodsTwelve New Zealand rabbits had transepithelial PRK. Six rabbits received topical recombinant human HGF (rhHGF) (50 µL of 0.1 mg/mL) 3 times a day for 1 week beginning 6 hours prior surgery and until full closure of the epithelium, and 6 control rabbits received vehicle by the same schedule. Slit lamp photos were taken immediately and at 43 to 45 hours after surgery to determine the rate of epithelial healing. Slit lamp photographs and immunohistochemistry for α-smooth muscle actin were analyzed at 1 month in masked fashion.ResultsThe rhHGF group tended to have slower re-epithelization when compared with the controls, but no statistically significant difference was noted (P = 0.62). There was no significant difference in the density of myofibroblasts in the central stroma (P = 0.49) or corneal opacity (P = 0.84) between the HGF and control groups at 1 month after PRK.ConclusionsTopical rhHGF applied three times a day during the early postoperative period prior to epithelial closure did not significantly change the corneal epithelial healing rate, myofibroblast density, or opacity compared with vehicle after transepithelial −9.0 D PRK injury of the central cornea in rabbits.Translational RelevanceHGF has been reported to decrease myofibroblast generation and fibrosis in many organs, but topical HGF applied to the cornea until epithelial healing had no effect on scarring fibrosis in rabbit corneas.

Combination of HGF and IGF-1 promotes connexin 43 expression and improves ventricular arrhythmia after myocardial infarction through activating the MAPK/ERK and MAPK/p38 signaling pathways in a rat model.

In this study, we hypothesized that the combination of hepatocyte growth factor (HGF) and insulin-like growth factor-1 (IGF-1) alters the expression of connexin 43 (Cx43) and results in a reduced frequency of induced ventricular arrhythmia in rats after myocardial infarction (MI) and explored the preliminary mechanisms involved. Cardiomyocytes were cultured in vitro in medium with PBS, HGF, IGF-1, GFs (HGF + IGF-1), HGF + p38 inhibitor, HGF + ERK inhibitor, IGF-1 + p38 inhibitor or IGF-1 + ERK inhibitor. The expression of Cx43 was tested by real-time PCR and Western blotting after 48 hours. MI was induced in 48 male Sprague-Dawley rats. The rats were randomly divided into four groups and received an injection of PBS, HGF, IGF-1 or GFs into the infarct border zone two weeks after MI. Six weeks after injection, the expression levels of Cx43 and programmed stimulation-induced ventricular arrhythmias were examined. In vitro, the expression of Cx43 mRNA and the Cx43 protein in cardiomyocytes was higher in the HGF, IGF-1, and GFs groups than in the PBS group. GFs had a combinatorial effect on the Cx43 mRNA level but not on the Cx43 protein level. There was a significant reduction in Cx43 mRNA and Cx43 protein levels in the IGF-1 + p38 inhibitor group and IGF-1 + ERK inhibitor group compared to the IGF-1 group. In vivo, programmed stimulation significantly decreased the frequency of ventricular arrhythmia in the GFs, HGF and IGF-1 groups, and this effect was accompanied by increased immunohistochemical staining for Cx43, myocardial Cx43 protein levels and Cx43 mRNA levels in the infarct border zone of the left ventricle compared with those in the PBS group. The combinatorial effect of GFs on Cx43 expression was only observed at the mRNA level. Both HGF and IGF-1 enhanced the expression of Cx43 and improved induced ventricular arrhythmia in rats with MI. Both synergistic and antagonistic effects of HGF and IGF-1 were not observed. In addition, IGF-1 may function through the MAPK/p38 and ERK1/2 signaling pathways to regulate Cx43 expression.

Up-regulation of Grb2-associated binder 1 promotes hepatocyte growth factor-induced endothelial progenitor cell proliferation and migration.

ObjectivesGrb2-associated binder 1 (Gab1), a scaffolding adaptor protein, plays an important role in transmitting key signals that control cell growth, migration, and function from multiple tyrosine kinase receptors. This study was designed to investigate the influence of upregulation of Gab1 in endothelial progenitor cells (EPCs) stimulated with hepatocyte growth factor (HGF), and the underlying molecular mechanisms.Materials and MethodsEndothelial progenitor cells isolated from human umbilical cord blood were identified and divided into four groups. EPCs in the Control group were cultured normally; those in the Control+HGF group were treated with HGF stimulation; those in the AD-Gab1 group were transfected with adenovirus containing the Gab1 gene but not treated with HGF stimulation; and, those in the AD-Gab1+HGF group were treated with both HGF stimulation and transfection with adenovirus containing the Gab1 gene. Subsequently, Gab1 expression and proliferation and migration ability were compared for EPCs grown under different conditions. Furthermore, we measured phosphorylation levels of three key proteins Gab1, SHP2, and ERK1/2.ResultsThe AD-Gab1+HGF group had the highest expression of Gab1 and higher proliferation and migration than the other three groups.ConclusionsUpregulation of Gab1 promoted HGF-induced EPC proliferation and migration. Mechanistically, HGF stimulated Gab1 tyrosine phosphorylation in EPCs, thus leading to activation of extracellular regulated MAP kinase 1/2, which is involved in proliferation and migration signaling.

Downregulation of miR-1224 protects against oxidative stress-induced acute liver injury by regulating hepatocyte growth factor.

To study the effect of microRNA-1224 (miR-1224) on hydrogen peroxide (H2 O 2 )-induced oxidative stress injury in hepatocytes, and explore its underlying mechanism. L02 cells were treated with H2 O 2 (100 mmol/L) to establish the model of an oxidative stress injury in hepatocytes. Quantitative reverse transcriptase polymerase chain reaction was used to detect the expression of miR-1224 and hepatocyte growth factor (HGF) in L02 cells. L02 cells were transfected with anti-miR-con (H 2 O 2 + anti-miR-con group), anti-miR-1224 (H 2 O 2 + anti-miR-1224 group), pcDNA3.1 (H 2 O 2 + ctrl group), pcDNA3.1-HGF (H 2 O 2 + HGF group), si-HGF and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + HGF group), si-NC and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + ctrl group) by liposome method. Cells without any treatment were regarded as a negative control (NC) group. The protein expression of HGF in each group cells was detected by Western blot analysis. Cell viability and apoptosis of each group were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay or flow cytometry, respectively. The interaction between miR-1224 and HGF was measured by dual luciferase reporter gene assay. The expression of miR-1224 was enhanced in H2 O 2 -treated L02 cells and its knockdown alleviated H 2 O 2 -induced suppression of viability and promotion of apoptosis. HGF is a target of miR-1224 and its overexpression abated H 2 O 2 -induced injury in hepatocytes. Moreover, silencing of HGF rescued the effect of downregulation of miR-1224 on cell viability and apoptosis in H 2 O 2 -treated L02 cell. Downregulation of miR-1224 could attenuate oxidative stress-induced inhibition of viability and increase of apoptosis in hepatocytes by targeting HGF, which may provide a target for potential therapy of acute liver injury.

Effects of three inducing factors on differentiation of bone marrow derived mesenchymal stem cells into lymphatic endothelial cells

Objective To observe the effects of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor C (VEGF-C) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into lymphatic endothelial cells (LECs). Methods The third to the fifth passage of BMSCs of rats were collected for the following experiments. (1) BMSCs of rats were collected and divided into negative control group, CD90 group, CD44 group, and CD34 group according to the random number table (the same grouping method below), with 3 samples in each group. Phosphate buffer of 5 μL was added to cells in negative control group, and cells in the other 3 groups were added with 5 μL corresponding antibodies respectively. The positive expression of cell surface antigen was detected by flow cytometer. (2) BMSCs of rats in 3 batches were collected and divided into blank control group, VEGF-C group, HGF group, bFGF group, VEGF-C+ HGF group, VEGF-C+ bFGF group, HGF+ bFGF group, and VEGF-C+ HGF+ bFGF group, with 3 samples in each group. Cells in blank control group were added with 2 mL complete medium, cells in VEGF-C group were added with 2 mL complete medium and 10 μL VEGF-C of 10 μg/mL, cells in HGF group were added with 2 mL complete medium and 16 μL HGF of 10 μg/mL, and cells in bFGF group were added with 2 mL complete medium and 20 μL bFGF of 1 μg/mL. Cells in VEGF-C+ HGF group, VEGF-C+ bFGF group, HGF+ bFGF group, and VEGF-C+ HGF+ bFGF group were added with 2 mL complete medium and induction factors with corresponding concentration and volume as above. On 10 d of culture, the morphology of the cells was observed by the inverted phase contrast microscope, and the protein and mRNA expressions of lymphatic vessel endothelial hyaluronic acid receptor 1 (LYVE-1), VEGF receptor 3 (VEGFR3), and integrin α9 were detected by Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction respectively. (3) BMSCs of rats were collected and divided into blank control group, HGF+ VEGF-C+ bFGF group, bFGF+ VEGF-C+ HGF group, and VEGF-C+ HGF+ bFGF group, with 3 samples in each group. Cells in blank control group were added with 2 mL complete medium. Cells in HGF+ VEGF-C+ bFGF group were added with 2 mL complete medium, 16 μL HGF of 10 μg/mL, and 10 μL VEGF-C of 10 μg/mL, after 6 hours, 20 μL bFGF of 1 μg/mL was added. Cells in bFGF+ VEGF-C+ HGF group were added with 2 mL complete medium, 20 μL bFGF of 1 μg/mL, and 10 μL VEGF-C of 10 μg/mL, after 6 hours, 16 μL HGF of 10 μg/mL was added. Cells in VEGF-C+ HGF+ bFGF group were simultaneously added with 2 mL complete medium and the same concentration and volume of three inducing factors as above. In addition, BMSCs of rats in another 2 batches were collected and grouped, and they were dealt with the same methods as above except that the interval time of 6 hours in HGF+ VEGF-C+ bFGF group and bFGF+ VEGF-C+ HGF group was adjusted to 12 and 24 hours. On 10 d of culture, protein expressions of LYVE-1, VEGFR3, and integrin α9 were detected by Western blotting. Data were processed with analysis of variance of factorial design, one-way analysis of variance, and least significant difference t test, and Bonferroni correction. Results (1) The positive expression rates of surface antigen of cells in negative control group, CD90 group, CD44 group, and CD34 group were 0.39%, 99.84%, 99.90%, and 0.57%, respectively. (2) On 10 d of culture, cells in blank control group, HGF group, bFGF group, and HGF+ bFGF group presented long fusiform, while cells in the other groups presented polygonal shape. (3) On 10 d of culture, there were no protein expressions of LYVE-1, VEGFR3, and integrin α9 in cells of blank control group, HGF group, bFGF group, and HGF+ bFGF group. On 10 d of culture, protein expressions of LYVE-1, VEGFR3, and integrin α9 in cells of VEGF-C+ HGF+ bFGF group were significantly higher than those in VEGF-C group (t=24.21, 11.04, 15.43, P 0.05). Conclusions VEGF-C is a necessary factor for inducing BMSCs to differentiate into LECs. HGF and bFGF may promote the differentiation by up-regulating the expressions of integrin α9 and VEGFR3 respectively. But the induction effects of the two factors may be independent. The combination of VEGF-C, HGF, and bFGF have the best effects of promoting differentiation. Key words: Mesenchymal stem cells; Vascular endothelial growth factor C; Hepatocyte growth factor; Cell differentiation; Basic fibroblast growth factor; Lymphatic endothelial cells

Effect of serum HGF and CK18 in patients with laryngeal cancer on postoperative prognosis

Objective: To investigate the correlation between the levels of serum cytokeratin 18 (CK18) and hepatocyte growth factor (HGF) and tumor stage and prognosis in patients with laryngeal carcinoma. Method: Patients with laryngeal cancer who were admitted to our hospital from January 2012 to December 2013 were enrolled. Blood samples were collected for CK18 and HGF detection, and postoperative prognosis data were collected at regular follow-up. Result: The serum CK18 and HGF concentration of patients with poorly differentiated tumor, high tumor grade and clinical stage were significantly higher than those with well differentiated tumor, low tumor grade and low clinical stage (P<0.05). The 5-year survival rate (69.9%) of patients with laryngeal cancer in the high CK18 group was significantly lower than that in the low CK18 group (92.3%) (P<0.05), and the 5-year survival rate (71.1%) of patients with laryngeal cancer in the high HGF group was significantly lower than that in the low HGF group (94.2%) (P<0.05). The results of univariate analysis showed that tumor type, differentiation, T grade, N grade and clinical stage affected the survival rate of patients with laryngeal cancer. Cox regression model analysis showed high CK18 and high HGF levels were risk factors for poor prognosis ［CK18 HR: 2.594 (1.558-4.318); P<0.001, HGF HR: 2.671 (1.605-4.446); P<0.001］. Conclusion:The results suggested that serum CK18 and HGF levels can be used as prognostic and disease monitoring biomarkers for laryngeal cancer.

Therapeutic effect of hepatocyte growth factor-overexpressing bone marrow-derived mesenchymal stem cells on CCl4-induced hepatocirrhosis.

Hepatocirrhosis is one of the most severe complications of chronic hepatic disease in terms of medical intervention, and the available therapies are limited and not very successful. In this study, bone marrow-derived mesenchymal stem cells (BM-MSCs) from host rats were transduced with an adenoviral vector labelled with green fluorescent protein (EGFP) to overexpress hepatocyte growth factor (HGF). The therapeutic effect of these modified stem cells (HGF-BM-MSC group) transplanted intravenously into hepatocirrhosis model rats treated with CCl4 was evaluated using serological, biochemical and histological approaches. We compared the rats in the HGF-BM-MSC group with those in the other groups (rats treated with BM-MSCs, rats treated with HGF and untreated rats (Controls)) in detail. The localisation of EGFP-tagged BM-MSCs in the injured liver was evaluated using a microscope, and the cells co-expressed hepatocyte nuclear factor 4α, albumin and cytokeratin 18. After treatment for 4 weeks, the HGF-BM-MSC, BM-MSC and HGF groups exhibited increased protein and mRNA levels of hepatocyte nuclear factor 4α, albumin and cytokeratin 18, but decreased levels of aspartate aminotransferase, alanine aminotransferase and total bilirubin. These findings indicate that BM-MSC transplantation and HGF application have great potential for the treatment of hepatocirrhosis.