FGF-2 Levels Research Articles

Coeloglossum viride var. bracteatum extract attenuates staurosporine induced neurotoxicity by restoring the FGF2-PI3K/Akt signaling axis and Dnmt3

We previously demonstrated the antioxidant activity of Coeloglossum viride var. bracteatum extract (CE) in rat cortical neurons and in mice with chemically induced cognitive impairment. In this work, we established a staurosporine (STS)-induced toxicity model to decipher the neuroprotective mechanisms of CE. We found that CE protected cell viability and neurite integrity in STS-induced toxicity by restoring the levels of FGF2 and its associated PI3K/Akt signaling axis. LY294002, a pan-inhibitor of PI3K, antagonized the activity of CE, although its-mediated restoration of FGF2 was unaffected. In addition, CE restored levels of Bcl-2/Caspase-3, PKCα/CaM pathway, and Dnmt3a and Dnmt3b, two methyltransferases that contribute to de novo DNA methylation. The Dnmts inhibitor 5-azacytidine impaired CE-mediated restoration of Dnmt3 or CaM, as well as the transition of DNA methylation status on the Dnmt3 promoter. These results reveal potential mechanisms that could facilitate the study and application of CE as a neuroprotective agent.

Tear cytokine profiles in patients with extranodal marginal zone B-cell lymphoma of the ocular adnexa.

To comprehensively analyse the tear cytokine levels of patients with extranodal marginal zone B-cell lymphoma (EMZL) of the ocular adnexa (OA), and the association with clinical characteristics. Tear cytokine concentrations of 21 OA-EMZL patients and 14 age- and sex-matched healthy controls were measured using a 27-multiplex bead analysis on a Luminex system. Tear break-up time, corneal fluorescent staining and other clinical and demographic data were collected as well. The diagnosis of OA-EMZL was established based on the incisional biopsy and histopathology. The concentrations of interleukin-1 receptor antagonist (IL-1RA) and IL-8, and the ratio of IL-1RA/IL-1β were significantly increased in OA-EMZL tear samples (all P < 0.05), while the levels of three cytokines (FGF-2, IL-2 and IL-4), as well as IL-10/IL-6 ratio were significantly decreased (all P < 0.05). The American Joint Committee on Cancer Tumour stage was significantly associated with tear concentrations of FGF-2 (r = -0.44, P = 0.043), GM-CSF (r = -0.49, P = 0.025) and IL-2 (r = -0.45, P = 0.042), while lacrimal gland lymphoma invasion was related to levels of IL-8 (r = 0.53, P = 0.012), FGF-2 (r = -0.43, P = 0.049) and IL-10/IL-6 ratio (r = -0.48, P = 0.026). Receiver operating characteristic (ROC) curve analysis revealed moderate diagnostic accuracy of these indices in differentiating OA-EMZL from normal eyes (area under ROC: 0.69-0.74). Multiple tear cytokines were significantly dysregulated in OA-EMZL patients. These cytokines could potentially serve as diagnostic biomarkers and therapeutic targets in future.

Intra-Individual Changes in Immune Marker Concentrations and Multiple Myeloma Risk: A Longitudinal Prospective Study Using Repeated Pre-Diagnostic Plasma Samples

Background: We studied immune marker trajectories (i.e. intra-individual changes in immune marker concentrations over time) of ten immune markers in relation to multiple myeloma (MM) risk. Low blood concentrations of MCP-3, MIP-1α, VEGF, FGF-2, Fractalkine, and TGF-α have been associated with myeloma risk in a previous study of single blood samples per participant collected around six years before diagnosis (Vermeulen et al., submitted). Longitudinal studies regarding these immune markers may provide substantial input to understanding their natural history in myeloma development and how these markers could be used in clinical routine.Methods: We identified 66 MM cases who had donated two blood samples before myeloma diagnosis, as well as matched cancer-free controls from the Northern Sweden Health and Disease Study. Blood levels of MCP-3, MIP-1α, MIP-1β, VEGF, FGF-2, Fractalkine, TGF-α, IL-13, TNF-α, and IL-10 were assessed in duplicate using Luminex bead-based multiplex assays (Millipore, Billerca, MA). Samples from matched cases and controls, as well as repeated samples from the same individuals, were included in random order within the same analytical batch. Linear mixed effects modeling was applied to investigate differences in immune marker trajectories between cases and controls.Findings: Samples were donated on median 12.3 ± 4.7 and 4.7 ± 3.8 years (± SD) prior to MM diagnosis for the baseline and the repeated sample, respectively. Median age at myeloma diagnosis was 62.9 ± 8.6 years (± SD). MM cases had significantly lower plasma levels of MCP-3, VEGF, FGF-2, Fractalkine, and TGF-α compared to controls. Notably, these immune markers also decreased significantly over time, while remaining temporally stable among control individuals. Decreasing marker trajectories were especially pronounced for VEGF (β = -0.012, P= 3.450 x 10-4), FGF-2 (β = -0.010, P= 5.700 x 10-5), and TGF-α (β = -0.019, P= 3.640 x 10-4) among future myeloma patients the closer to diagnosis the sample was donated. Subgrouping of samples by time between blood draw and diagnosis showed that plasma levels of MCP-3 were inversely associated with myeloma risk in samples collected as long as 12 to 24 years before diagnosis (OR = 0.23, Ptrend = 0.036) (OR = 4th vs. 1st quartile).Interpretation: Our data provide further support of inverse associations between MM risk and plasma levels of MCP-3, VEGF, FGF-2, Fractalkine, and TGF-α. Low MCP-3 levels might be involved in early myeloma development. Considering that monoclonal gammopathy of unclear significance (MGUS) is detectable in the majority of myeloma patients in samples collected up to 15 years before diagnosis (Landgren et. al. Blood 2009, Weiss et. al. Blood 2009), decreasing immune marker levels of VEGF, FGF-2, and TGF-α might be indicative of progression to MM among MGUS patients. Therefore, these markers could be of potential use as biomarkers to improve prediction of myeloma progression in patients followed for MGUS. However, larger studies including samples from both, myeloma and MGUS patients together with matched controls are needed to further evaluate this hypothesis. DisclosuresNo relevant conflicts of interest to declare.

Association of circulating fibroblast growth factor-2 with progression of HIV-chronic kidney diseases in children.

Children living with HIV frequently show high plasma levels of fibroblast growth factor-2 (FGF-2/bFGF). FGF-2 accelerates the progression of several experimental kidney diseases; however, the role of circulating FGF-2 in childhood HIV-chronic kidney diseases (HIV-CKDs) is unknown. We carried out this study to determine whether high plasma FGF-2 levels were associated with the development of HIV-CKDs in children. The plasma and urine FGF-2 levels were measured in 84 children (< 12 years of age) living with HIV during the pre-modern antiretroviral era, and followed for at least 3 years to determine the prevalence of proteinuria and HIV-CKDs. We also assessed the distribution of the kidney FGF-2 binding sites by autoradiography and Alcian blue staining, and explored potential mechanisms by which circulating FGF-2 may precipitate HIV-CKDs in cultured kidney epithelial and mononuclear cells derived from children with HIV-CKDs. High plasma FGF-2 levels were associated with a high viral load. Thirteen children (~ 15%) developed HIV-CKDs and showed a large reservoir of FGF-2 low-affinity binding sites in the kidney, which can facilitate the recruitment of circulating FGF-2. Children with high plasma and urine FGF-2 levels had 73-fold increased odds (95% CI 9-791) of having HIV-CKDs relative to those with normal FGF-2 values. FGF-2 induced the proliferation and decreased the expression of APOL-1 mRNA in podocytes, and increased the attachment and survival of infected mononuclear cells cultured from children with HIV-CKDs. High plasma FGF-2 levels appear to be an additional risk factor for developing progressive childhood HIV-CKDs.

Integrated Analysis of mRNA and miRNA Changes in Two Haliotis diversicolor Genotypes and Their Hybrid

Heterosis is a widely distributed phenomenon in mollusks. It is vital in aquaculture by bringing beneficial traits into hybrids. People have utilized the heterosis theory in aquaculture for years. However, the molecular basis of heterosis remains elusive. Evident growth and survival heterosis were shown in the hybrid (“Dongyou-1”) of two Haliotis diversicolor geographic genotypes (Japan and Taiwan). To explore the molecular basis underlying the hybrid abalone’s heterosis, we conducted comparative mRNA and miRNA transcriptional analysis in the hybrid and parental genotypes. Differentially expression analysis identified 5,562 differentially expressed genes (DEGs) and 102 differentially expressed miRNAs (DEMs) between the three genotypes. 1,789 DEGs and 71 DEMs were found to be non-additively expressed in the hybrid. Meanwhile, both the expression level dominance pattern (ELD) and expression level overdominance pattern (ELOD) were found in the DEGs and DEMs, showing the existence of dominance and overdominance models in the hybrid’s transcriptome and post-transcriptional regulation. Functional analysis showed the non-additively expressed genes, ELD genes, and ELOD genes were significantly enriched in growth, immunity, and stress response related pathways, while some of the pathways were regulated by the mRNA-miRNA interactions. The expression levels of FGF, C1Q, HC, CAT, SEGPX, and MGST were significantly up-regulated in the hybrid compared to the middle parent value. In conclusion, we identified the existence of non-additivity, dominance, and overdominance models in the transcriptome and miRNAome of the H. diversicolor hybrid; these models facilitate the advantageous parental alleles’ integration into the hybrid, contributing to the hybrid’s growth and survival heterosis.

MO793COMPARATIVE STUDY BETWEEN THE IMPACT OF ETELCALCETIDE AND CINACALCET ON LEVELS OF IPTH, FGF-23, KLOTHO PROTEIN, SCLEROSTIN, CA AND P IN DIALYSIS PATIENTS WITH SECONDARY HYPERPARATHYROIDISM. RESULTS FROM A PROSPECTIVE RANDOMIZED TRIAL

Abstract Background and Aims The treatment of secondary hyperparathyroidism is one of the main tasks in the correction of mineral and bone disorders (MBD) in patients with chronic kidney disease (CKD). However, the results of therapy for secondary hyperparathyroidism are still unsatisfactory. In our prospective randomized controlled trial were evaluated the effect and safety of 26 weeks of treatment with etelcalcetide (intravenous route of administration) compare with cinacalcet (oral administration) on CKD-MBD parameters in patients on program hemodialysis with secondary hyperparathyroidism. Method The study group included 50 stable patients receiving hemodialysis with secondary hyperparathyroidism (PTH &amp;gt; 300 pg/ml) and corrected Ca level greater than 2.2 mmol/L, who were randomized in a 1: 1 ratio for treatment with etelcalcetide (n = 25) or cinacalcet (n = 25) for 26 weeks. All patients were monthly evaluated the levels of P, Ca, intact parathyroid hormone (iPTH), alkaline phosphatase (ALP); the levels of FGF 23, Klotho protein and sclerostin were assessed once in 3 months. The dose of both drugs was adjusted according to the serum iPTH level. The nature, frequency, and severity of treatment-emergent adverse events were assessed. Results Therapy with cinacalcet and etelcalcetide led to a significant decrease in the level of iPTH in the blood serum from 613.1 ± 235.1 to 302.2 ± 205.1pg/ml (p&amp;lt; 0.01) and from 671.2 ± 272.3 to 358,6 ± 292.5 pg/ml (p &amp;lt;0.01), by 49.2% and 53.4%, respectively. A significant decrease in the levels of corrected Ca was noted in both groups: in the etelcalcetide group from 2.20 ± 0.12 to 2.06 ± 0.18 mmol/L (p &amp;lt;0.05), in the cinacalcet group from 2.25 ± 0.12 to 2.04 ± 0.21 mmol/l (p &amp;lt;0.05). There was no significant change in the P levels. The alkaline phosphatase level significantly decreased in the cinacalcet group from 178.7 ± 116.8 to 78.9 ± 34.1 U/L, p &amp;lt;0.05) and in the etelcalcetide group from 170,3 ± 115.7 to 87.1 ± 30.8 U/L, p &amp;lt;0.05. There was a significant increase in Klotho protein levels by the end of the study from 17.9 ± 5.0 to 57.1 ± 39.3 (p &amp;lt;0.05) and from 17.6 ± 3.7 to 91.6 ± 56.2 pg/ml (p &amp;lt;0.05), respectively, in the cinacalcet and etelcalcetide group. Changes in FGF-23 and sclerostin by 6 months reached statistically significant changes only in the etelcalcetide group, a decrease from the FGF-23 level from 42.7 ± 22.2 to 23.0 ± 12.3 pg/ml and an increase in the level of sclerostin from 1, 59 ± 0.31 to 2.20 ± 0.33 ng/ml (p &amp;lt;0.05). During the study, 2 patients in the cinacalcet group dropped out due to dyspeptic symptoms and 1 patient in the etelcalcetide group dropped out due to hypocalcemia. Conclusion Etelcalcetide and cinacalcet are effective PTH-lowering drugs with a comparable safety profile. Treatment with etelcalcetide, in contrast to cinacalcet, was associated with significant increases in sclerostin and decreases in FGF-23, which may have beneficial effects on outcomes and requires further study.

Toll-Like Receptor Signalling Pathways Regulate Hypoxic Stress Induced Fibroblast Growth Factor but Not Vascular Endothelial Growth Factor-A in Human Microvascular Endothelial Cells

Retinal diseases are the leading causes of irreversible blindness worldwide. The role of toll-like receptor (TLR) signalling mechanisms (MyD88 and TRIF) in the production of pro-angiogenic growth factors from human microvascular endothelial cells (HMEC-1) under hypoxic stress remains unexplored. HMEC-1 was incubated under normoxic (5% CO2 at 37 °C) and hypoxic (1% O2, 5% CO2, and 94% N2; at 37 °C) conditions for 2, 6, 24, and 48 h, respectively. For TLR pathway analysis, HMEC-1 was pre-treated with pharmacological inhibitors (Pepinh-MyD88 and Pepinh-TRIF) and subjected to normoxia and hypoxia conditions. Gene and protein expressions of vascular endothelial growth factor-A (VEGF-A), fibroblast growth factor (FGF-2), hypoxia inducible factor 1-alpha (HIF1-α) were performed using quantitative polymerase chain reaction (qPCR), ELISA, and Western blot methodologies. Levels of TLR3 and TLR4 were analysed by flow cytometry. Under hypoxia, levels of VEGF-A and FGF-2 were elevated in a time-dependent fashion. Inhibition of MyD88 and TRIF signalling pathways decreased FGF-2 levels but failed to modulate the secretion of VEGF-A from HMEC-1. Blocking a known regulator, endothelin receptor (ETR), also had no effect on VEGF-A secretion from HMEC-1. Overall, this study provides the proof-of-concept to target TLR signalling pathways for the management of blinding retinal diseases.

Effect of Endurance Training and Stem Cell on Fgf2 and Mmp13 Gene Expression in Knee Tissue of Rats with Osteoarthritis

Introduction: The aim of this study was to investigate the effect of endurance training and stem cell injection on FGF2 and MMP13 gene expression in knee tissue of rats with knee osteoarthritis. Methods: The type of study was an experimental one.Male Wistar rats were randomly divided into 5 groups: control - healthy, control - patient, patient - stem cell, patient - training, and patient - training - stem cell. The training program consisted of 30 minutes of running on a treadmill with no slope at 16 m / min for the first week, with 1 m / min added weekly. Rats received MSCs through intracellular injection of 1*106 cells / kg. Expression of FGF2 and MMP13 genes was measured by Real Time PCR. One way ANOVA and if there was a significant difference, Tukey post hoc test were used to determine the difference between groups. All statistical analyzes were performed using SPSS version 20. Results: Data analysis showed that training and stem cell therapy have significantly increased in genes expression of FGF2 and MMP-13 in mice with osteoarthritis (p &lt;0.000). Moreover, Tukey post hoc test showed a significant difference in the level of FGF2 changes in the training-stem cell groups compared to the training group and the saline group (p=0.000‌). There was also a significant difference between the MMP-13 follow-up test with the control group with the saline group, the training group and the stem cell with the train-stem cell, and the saline group with the training group9(P=0.000‌). Conclusion: The results showed that FGF2 levels and MMP13 in the cartilage tissue of mice with osteoarthritis have increased and endurance training and stem cells therapy caused a decrease in the level of factors.

Growth Factors Assessed during Kasai Procedure in Liver and Serum Are Not Predictive for the Postoperative Liver Deterioration in Infants with Biliary Atresia.

Background: Biliary atresia (BA) is a neonatal cholangiopathy characterized by progressive destruction of the biliary system resulting in liver cirrhosis. Residual bile drainage can temporarily be achieved through Kasai portoenterostomy (KPE) and some children show long-term survival with their native liver. However, most children eventually require liver transplantation (LTX). As several growth factors (GF) and chemokines have been shown to promote fibrogenesis in the liver, we assessed whether GF are predictive for the course of disease. Material and Methods: Liver and sera samples were collected from 49 infants with BA during KPE. Levels of 13 different GF were measured by multiplex immunoassay. Patient outcomes were stratified into favorable (bilirubin < 20 µmol/L at 2-year follow-up) and unfavorable (LTX). GF levels were compared between groups by a t-test, correlation coefficients were calculated, and principal component analyses performed. Results: Twenty-two patients showed a favorable and 27 an unfavorable disease course. No relation of GF and outcome could be established. In both groups, high levels of SDF-1alpha/CXCL12 (1473.0 ± 497.5 pg/mL), FGF2 (301.2 ± 207.8 pg/mL), and VEGF-a (209.0 ± 146.4 pg/mL) levels were measured within the liver, followed (in descending order) by PDGF-bb, LIF, GM-CSF, BDNF, VEGF-d, beta-NGF, IL-7, SCF, PIGF-1, and EGF. Serum marker levels showed much higher mean variation compared to hepatic values and no correlation to the protein microenvironment in the liver. Conclusions: Our study demonstrates high amounts of GF in livers from infants with BA at KPE, but no correlation to the outcome or serum values could be established. Our data suggest that local or systemic GF levels are unsuitable for prediction of the disease course. Collectively, we conclude that in BA the degree of proliferative activity caused by GF is a dismissible factor for the further course of disease.

The Role of Fgf9 in the Antidepressant Effects of Exercise and Fluoxetine in Chronic Unpredictable Mild Stress Mice.

The neurotrophic hypothesis of depression posits that stress and depression decrease neurotrophic factor expression in brain, whereas antidepressants and exercise can contribute to the blockade of stress effects and produce antidepressant effects. Fibroblast growth factor 9 (FGF9), a member of the fibroblast growth factor (FGF) family, has been reported to be dysregulated in depression. The present study aimed to determine whether and how Fgf9 mediates the antidepressant effects of fluoxetine and exercise in chronic unpredictable mild stress (CUMS) mice. Male C57BL/6 mice were exposed to CUMS for 7 weeks. From the fourth week, CUMS-exposed mice were subjected to fluoxetine treatment or swimming exercise for 4 weeks. Forced swim test, tail suspension test, and hole-board test were used to assess behaviors of mice. Real-time polymerase chain reaction was used to examine hippocampal messenger RNA levels of Fgf9, Fgf2, FgfR1, FgfR2, and FgfR3. Western blotting was used to examine the protein levels of Fgf9, protein kinase B (Akt), and phosphorylation of Akt at Ser473 in mouse hippocampus. Our results demonstrated that CUMS induced depression-like behaviors, which were reversed by fluoxetine treatment and swimming exercise. Moreover, we found that CUMS resulted in a dysregulation of Fgf9, Fgf2, and FgfR2 expression, whereas fluoxetine and swimming restored the FGF expression in CUMS-exposed mice. An analysis of the proteins suggests that the antidepressant effects of fluoxetine and exercise in CUMS-exposed mice were associated with ameliorated Fgf9/Akt signaling. Our findings have demonstrated that swimming exercise mimics the antidepressant effects of fluoxetine by regulating Fgf9 in CUMS-exposed mice, which may offer new mechanism-based therapeutic targets for depression.

S100A4 Is Involved in Stimulatory Effects Elicited by the FGF2/FGFR1 Signaling Pathway in Triple-Negative Breast Cancer (TNBC) Cells.

Triple-negative breast cancer (TNBC) is an aggressive breast tumor subtype characterized by poor clinical outcome. In recent years, numerous advancements have been made to better understand the biological landscape of TNBC, though appropriate targets still remain to be determined. In the present study, we have determined that the expression levels of FGF2 and S100A4 are higher in TNBC with respect to non-TNBC patients when analyzing “The Invasive Breast Cancer Cohort of The Cancer Genome Atlas” (TCGA) dataset. In addition, we have found that the gene expression of FGF2 is positively correlated with S100A4 in TNBC samples. Performing quantitative PCR, Western blot, CRISPR/Cas9 genome editing, promoter studies, immunofluorescence analysis, subcellular fractionation studies, and ChIP assays, we have also demonstrated that FGF2 induces in TNBC cells the upregulation and secretion of S100A4 via FGFR1, along with the ERK1/2–AKT–c-Rel transduction signaling. Using conditioned medium from TNBC cells stimulated with FGF2, we have also ascertained that the paracrine activation of the S100A4/RAGE pathway triggers angiogenic effects in vascular endothelial cells (HUVECs) and promotes the migration of cancer-associated fibroblasts (CAFs). Collectively, our data provide novel insights into the action of the FGF2/FGFR1 axis through S100A4 toward stimulatory effects elicited in TNBC cells.

The level of fibroblast growth factor-2 prepared from Advanced Platelet Rich Fibrin (A-PRF) in obese Saudi subjects compared to healthy subjects.

SummaryBackgroundThe prevalence of obesity has increased substantially in the last few decades. World Health Organization (2020) estimated that around 600 million obese adults worldwide were obese, and a further increase is expected in the future due to increased consumption of high-calorie diets and a sedentary lifestyle as per the evidence.AimTo evaluate and compare the level of fibroblast growth factor platelet rich fibrin (A-PRF) in obese subject compare to healthy weight subject.MethodsBlood samples were collected from 23 volunteers, 15 obese subject (test group) and 8 non-obese (control group) at Riyadh Elm University. Considering the smaller sample size of our study, the results are to cautiously be interpreted for generalizability. Studies employing larger sample size are recommended to overcome this point. But considering the meticulous study procedure adhering to the study protocol and set criteria, the study pronounces greater internal validity in the sample chosen. The medical, dental histories, an interview and clinical examination was performed to check the eligibility of the participants to be involved in this study, Blood sample was collected in 10 ml syringe, then being processed using A-PRF centrifugation protocols. Ten milliliters of whole blood without anticoagulant was centrifuged at 1,300 rpm for 14 minutes.ResultsThe level of FGF-2 released from (A-PRF) concentration was significantly lower on obese which was measured on 4 different times (day 1, day 7, day 14 and day 28), compared to healthy.ConclusionsThere was decrease in FGF-2 level released from (A-PRF) from obese compared to healthy.

The role of sperm in inducing genomic changes in the implantation: An experimental study.

Endometrial receptivity and implantation are important topics in reproductive sciences. No evidence was found to support sperm involvement in endometrial receptivity and its associated factors. This study aimed to explore the effect of the normal human spermatozoa-endometrium cell interaction in regulating genes in the endometrial receptivity pathway. Semen samples were collected from a healthy and fertile man; then, they were incubated with endometrial cells for 24hr and considered as the sperm group. A group was cultured without spermatozoa and considered as a control group. About 24hr later, cells were collected from the bottom of the culture dish. The expressions of the VEGF, FGF2, HBEGF, LIFR, EGF, LIF, MUC1, HOXA10, CSF and PGR genes were evaluated in the two groups. Statistical analysis was performed using an independent sample test. Compared with the control group, in the sperm group, the mRNA levels of PGR (p=.0451), VEGF (p=.0101), HBEGF (p=.0163), EFG (p=.0339), FGF2 (p=.012), LIF (p=.0324), LIFR (p=.0321) and HOXA10 (p=.0098) were significantly upregulated. The results showed that there is a need for the interaction between spermatozoa and endometrium for implantation and can be used for preparing uterine in in vitro fertilisation cycles.

It’s all about who you know: Memory retention of a rat’s cagemates during infancy negatively predicts adulthood hippocampal FGF2

Recent research has demonstrated that individual differences in infant fear memory positively predict adulthood anxiety-like behavior and conditioned fear expression. However, the physiological mechanisms underlying this relationship and the effect of environmental (e.g., social) influences on the stability of this relationship have not been explored. In the present study, we examined whether individual differences in infant fear memory predict levels of endogenous fibroblast growth factor-2 (FGF2; a biomarker of fear/anxiety) in adulthood, and whether the mean memory retention of a rat’s cagemates predicts conditioned fear expression and FGF2 in adulthood. We conditioned infant rats to associate a white noise with shock, and tested their memory of the association 1 week later. They were then weaned and randomly assigned to cage/cagemates. In adulthood, rats received weak context conditioning (i.e., a single shock) and were tested for fear of the context the following day. Rats were then euthanized and their brains extracted to measure levels of hippocampal FGF2 protein. Across 2 experiments, an individual rat’s fear memory during infancy positively predicted their own fear expression in adulthood, but the mean memory retention of their cagemates did not predict fear expression. In contrast, the mean memory retention of a rat’s cagemates during infancy negatively predicted hippocampal FGF2 protein in adulthood, but an individual rat’s memory retention did not predict their own levels of FGF2. These data support the idea that variations in the fearfulness of a rat’s cagemates predict individual differences on physiological measures in adulthood.

Conversion of Reactive Astrocytes to Induced Neurons Enhances Neuronal Repair and Functional Recovery After Ischemic Stroke.

The master neuronal transcription factor NeuroD1 can directly reprogram astrocytes into induced neurons (iNeurons) after stroke. Using viral vectors to drive ectopic ND1 expression in gliotic astrocytes after brain injury presents an autologous form of cell therapy for neurodegenerative disease. Cultured astrocytes transfected with ND1 exhibited reduced proliferation and adopted neuronal morphology within 2–3 weeks later, expressed neuronal/synaptic markers, and extended processes. Whole-cell recordings detected the firing of evoked action potentials in converted iNeurons. Focal ischemic stroke was induced in adult GFAP-Cre-Rosa-YFP mice that then received ND1 lentivirus injections into the peri-infarct region 7 days after stroke. Reprogrammed cells did not express stemness genes, while 2–6 weeks later converted cells were co-labeled with YFP (constitutively activated in astrocytes), mCherry (ND1 infection marker), and NeuN (mature neuronal marker). Approximately 66% of infected cells became NeuN-positive neurons. The majority (~80%) of converted cells expressed the vascular glutamate transporter (vGLUT) of glutamatergic neurons. ND1 treatment reduced astrogliosis, and some iNeurons located/survived inside of the savaged ischemic core. Western blotting detected higher levels of BDNF, FGF, and PSD-95 in ND1-treated mice. MultiElectrode Array (MEA) recordings in brain slices revealed that the ND1-induced reprogramming restored interrupted cortical circuits and synaptic plasticity. Furthermore, ND1 treatment significantly improved locomotor, sensorimotor, and psychological functions. Thus, conversion of endogenous astrocytes to neurons represents a plausible, on-site regenerative therapy for stroke.

Paeoniflorin exerts antidepressant-like effects through enhancing neuronal FGF-2 by microglial inactivation

Ethnopharmacological relevance Paeonia lactiflora is a famous Traditional Chinese medicine widely used for immunological regulation. Paeoniflorin, the main component of Paeonia lactiflora, exerts neuroprotective and antidepressant-like effects in rodents. Aim of the studyFibroblast growth factor 2 (FGF-2) is essentially required in the central nervous system as it acts as both a neurotrophic factor and an anti-inflammatory factor participating in the regulation of proliferation, differentiation and apoptosis of neurons in the brain. However, it is unclear whether paeoniflorin could exert antidepressant effects via regulating FGF-2. Materials and methodsIn the present study, the effects of paeoniflorin were evaluated in depressive mice induced by the endotoxin lipopolysaccharide (LPS) injection. ResultsThe results showed that paeoniflorin markedly increased sucrose preference and reduced immobility time in LPS mice, indicating antidepressant effects. Consistent with the results from molecular docking showing paeoniflorin antagonizes TLR4, NF-κB and NLRP3, the biochemical analysis also indicated paeoniflorin inhibited TLR4/NF-κB/NLRP3 signaling, decreased proinflammatory cytokine levels and microglial activation in the hippocampus of LPS induced mice. In addition, the levels of neuronal FGF-2 and the density of dendritic spine were improved by paeoniflorin. More importantly, the FGFR1 inhibitor SU5402 prevented the antidepressant effects of paeoniflorin and blocked the neuroinflammatory and neurogenic regulatory effects of paeoniflorin, indicating that FGF-2/FGFR1 activation was required for the effects of paeoniflorin. ConclusionTaken together, the results demonstrate that paeoniflorin exhibits neuroprotective and antidepressant effects in mice, which may be mediated by activating neuronal FGF-2/FGFR1 signaling via the inhibition of microglial activation in the hippocampus.

FGF gene expression in injured tendons as a prognostic biomarker of 1-year patient outcome after Achilles tendon repair

PurposeHealing outcome after Achilles Tendon Rupture (ATR) is variable and unsatisfactory. Many ATR patients still exhibit pain, functional deficits and limitations in walking one-year post-surgery. The present study was designed to investigate the association between the expression of healing biomarkers and patient outcome after ATR.MethodsTendon biopsies were collected from 25 ATR patients during surgery. At 1-year post surgery, all patients completed questionnaires; Achilles tendon Total Rupture Score (ATRS) and Foot and Ankle Outcome Score (FAOS), and were tested for functional outcomes by heel-rise test. In biopsies, FGF, COL III, FN, COL I and MMP-9 mRNA levels were assessed by quantitative RT-PCR while protein expression was studied by immunohistochemistry (IHC).ResultsOur analysis confirmed the presence of FGF, COL III, FN, COL I and MMP-9 at mRNA and protein levels in tendon biopsies. FGF gene expression associated positively with improved total ATRS and better functional outcomes. Additionally, FGF mRNA levels were associated with less pain, less running limitations and less loss in physical activity. In addition, higher COL III mRNA expression was associated with more tendon strength.ConclusionOur findings indicate that FGF gene expression is associated with improved patient-reported outcome. FGF expression in surgical biopsies could potentially be used to assist the prognostic evaluation of patient outcome and may be used as a predictor for healing. However, further studies are needed to evaluate the role of FGF in Achilles tendon healing.Level of evidenceII

Effects of a Catechol-Functionalized Hyaluronic Acid Patch Combined with Human Adipose-Derived Stem Cells in Diabetic Wound Healing.

Introduction: Chronic inflammation and impaired neovascularization play critical roles in delayed wound healing in diabetic patients. To overcome the limitations of current diabetic wound (DBW) management interventions, we investigated the effects of a catechol-functionalized hyaluronic acid (HA-CA) patch combined with adipose-derived mesenchymal stem cells (ADSCs) in DBW mouse models. Methods: Diabetes in mice (C57BL/6, male) was induced by streptozotocin (50 mg/kg, >250 mg/dL). Mice were divided into four groups: control (DBW) group, ADSCs group, HA-CA group, and HA-CA + ADSCs group (n = 10 per group). Fluorescently labeled ADSCs (5 × 105 cells/100 µL) were transplanted into healthy tissues at the wound boundary or deposited at the HA-CA patch at the wound site. The wound area was visually examined. Collagen content, granulation tissue thickness and vascularity, cell apoptosis, and re-epithelialization were assessed. Angiogenesis was evaluated by immunohistochemistry, quantitative real-time polymerase chain reaction, and Western blot. Results: DBW size was significantly smaller in the HA-CA + ADSCs group (8% ± 2%) compared with the control (16% ± 5%, p < 0.01) and ADSCs (24% ± 17%, p < 0.05) groups. In mice treated with HA-CA + ADSCs, the epidermis was regenerated, and skin thickness was restored. CD31 and von Willebrand factor-positive vessels were detected in mice treated with HA-CA + ADSCs. The mRNA and protein levels of VEGF, IGF-1, FGF-2, ANG-1, PIK, and AKT in the HA-CA + ADSCs group were the highest among all groups, although the Spred1 and ERK expression levels remained unchanged. Conclusions: The combination of HA-CA and ADSCs provided synergistic wound healing effects by maximizing paracrine signaling and angiogenesis via the PI3K/AKT pathway. Therefore, ADSC-loaded HA-CA might represent a novel strategy for the treatment of DBW.

Effects of metformin on lipopolysaccharide induced inflammation by activating fibroblast growth factor 21

ABSTRACT Lipopolysaccharide (LPS) is a component of the cell wall of Gram-negative bacteria that produces endotoxemia, which may cause septic shock. Metformin (MET) is a widely used hypoglycemic drug that exhibits anti-inflammatory properties. Fibroblast growth factor 21 (FGF21) is an endocrine polypeptide that affects glucose and lipid metabolism, and also possesses anti-inflammatory properties. We investigated the effects of MET and FGF21 on inflammation due to LPS induced endotoxemia in male rats. Animals were divided into five groups: control, LPS, pre-MET LPS, LPS + 1 h MET and LPS + 3 h MET. Serum levels of alanine aminotransferase, aspartate aminotransferase, FGF2, interleukin-10 and tumor necrosis factor alpha were measured. Malondialdehyde, myeloperoxidase and FGF21 levels were measured in liver tissue samples. Histopathology of all groups was assessed using hematoxylin and eosin stained sections. LPS caused severe inflammatory liver damage. MET exhibited a partially protective effect and reduced inflammation significantly. FGF21 is produced in the liver following inflammation and MET may increase its production.

Co-transfection with BMP2 and FGF2 via chitosan nanoparticles potentiates osteogenesis in human adipose-derived stromal cells in vitro.

ObjectiveTo investigate if co-transfection of human bone morphogenetic protein 2 (BMP-2, BMP2) and human fibroblast growth factor 2 (FGF2, FGF2) via chitosan nanoparticles promotes osteogenesis in human adipose tissue-derived stem cells (ADSCs) in vitro.Materials and MethodsRecombinant BMP2 and/or FGF2 expression vectors were constructed and packaged into chitosan nanoparticles. The chitosan nanoparticles were characterized by atomic force microscopy. Gene and protein expression levels of BMP-2 and FGF2 in ADSCs in vitro were evaluated by real-time polymerase chain reaction (PCR), western blot, and enzyme-linked immunosorbent assay. Osteocalcin (OCN) and bone sialoprotein (BSP) gene expression were also evaluated by real-time PCR to assess osteogenesis.ResultsThe prepared chitosan nanoparticles were spherical with a relatively homogenous size distribution. The BMP2 and FGF2 vectors were successfully transfected into ADSCs. BMP-2 and FGF2 mRNA and protein levels were significantly up-regulated in the co-transfection group compared with the control group. OCN and BSP mRNA levels were also significantly increased in the co-transfection group compared with cells transfected with BMP2 or FGF2 alone, suggesting that co-transfection significantly enhanced osteogenesis.ConclusionsCo-transfection of human ADSCs with BMP2/FGF2 via chitosan nanoparticles efficiently promotes the osteogenic properties of ADSCs in vitro.