Basic fibroblast growth factor promoting the healing of palatal perforation in rats

AIM: To study the influence of basic fibroblast growth factor(bFGF) on treating ed palatal perforation in rats. METHODS: bFGF was given to the early palatal perforation in rat. The granulation tissues in perforations were grossly and pathologically obserVed. RESULTS: The the of wound healing was significantly in- crease in the bFGF group (P<0.01 ). bFGF could obviously promote the proliferation of the granulation tissues and fibroblasts. The number of AgNORs granules in fibrablasts were 3.73 ±0.52 in the buy group and 2.11 ±0.31 in control group (P < 0.05). CONCLUSION: bFGF could promote the growth of granulation tissues and had a strong promotion on the wound healing in palatal perforation. It was helpful in repairing palatal perforation.     

The condition for the healing of dural defects in vitro and the influential factors

AIM:To study the condition for the healing process of dural defects in vitro and its influential factors. METHODS:Rabbit dura mater was cultured in 24-well plates that had been coated with collagen, laminin and polylysine to observe the influence of extracellular matrix on it. Dural cells were subcultured on slides for immunocytochemistry. Dural healing was observed by scanning electronic microscope. Different growth factors were added to the culture medium respectively to analyze the influential factors on it. RESULTS:Only the dura pieces cultured on collagen coated wells showed migration of cells into the central defect. Dural cells stained strongly positive with antibodies against vimentin and negative with Ⅷ factor. The dural pieces with bFGF added showed earlier migration of dural cells from defect margin after a period of 3-4 days and the dural defect healing occurred after 7-8 days. CONCLUSION:Collagen is an essential matrix for the dural healing. Cellular migration from the defect margin is an important echanism in the process of dural healing and bFGF accelerates dural healing.     

Expression of VEGF and bFGF in rat model of pressure ulcer

AIM:To study the effects of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) on the molecular pathogenesis of pressure ulcer.METHODS:SD rats were randomly divided into control group and experiment group. The pressure ulcer model was established by magnetic disk circulating compression method. HE staining was used to observe the pathological changes of the skin in the rats. The expression of VEGF and bFGF in the tissues was detected by immunohistochemical method. RESULTS:The expression of VEGF and bFGF in the tissues of rat Ⅲ-degree pressure ulcer was lower than that in the surrounding tissues and normal skin(P<0.01). The changes of VEGF and bFGF were consistent(κ=0.58). CONCLUSION:The expression levels of VEGF and bFGF are decreased in the tissues of rat pressure ulcer, suggesting that they may be the potential key factors in the difficult healing of pressure ulcer.     

Involvement of bFGF in the lung response to silica in a mouse model

AIM: To investigate the relationship between basic fibroblast growth factor (bFGF) and the development of silicosis in mice. METHODS: MTT test was utilized to examine the effects of bFGF-neutralizing antibody and the bronchoalveolar lavage fluid (BALF) of the mice exposed to silica on lung fibroblast cell growth. RESULTS: BALF from mice treated intrabronchially with silica promoted the growth of lung fibroblasts and anti-bFGF antibody inhibited the effect of BALF dramatically. CONCLUSION: These results indicates that bFGF secretion increases in lung in a mice silicosis model and participates in the development of silicosis.     

Effects of angiotensin II type 2 receptor blocker on wound healing in a mouse skin full-thickness injury model

AIM：To observe the effect of angiotensin II (Ang II) and its type 2 receptor (AT2R) on re-epithelialization, granulation tissue formation and growth factor production during wound healing, and to explore the possible mechanism by which Ang II and AT2R influence wound healing. METHODS：Two full-thickness skin wounds were created on the dorsum of C57BL/6J mice. The animals were treated with or without AT2R blocker PD123319 at a dose of 10 mg/kg daily after wounding. Specimens were taken from the wound of each mouse on days 3, 5, 7, 9, 11, 13 and 15 after wounding. Re-epithelialization and granulation tissue formation in the wounded skin tissues were evaluated by HE staining. The production of growth factors,epidermal growth factor (EGF),vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in wounded tissues during the healing process was detected by ELISA. RESULTS：Treatment with PD123319 significantly increased the rate of re-epithelialization and the area of granulation tissue compared with control group at 5 d and 7 d after wounding. Moreover, peritoneal application of PD123319 increased the production of EGF, VEGF and bFGF in the wounded tissues at the indicated time points after wounding. CONCLUSION：AT2R blocker PD123319 accelerates wound healing via promoting re-epithelialization,granulation tissue formation and growth factor production.     

Effects of basic fibroblast growth factor on proliferation and collagen production in human umbilical cord mesenchymal stem cells

AIM：To observe the growth pattern and surface markers of human umbilical cord mesenchymal stem cells(hUCMSCs) and to explore the influence of basic fibroblast growth factor (bFGF) on the proliferation and collagen production in hUCMSCs cultured in vitro. METHODS：hUCMSCs were isolated by enzyme digestion method and adherent culture. The surface markers CD45, CD34, CD105, CD29 and HLA-DR of the cells were analyzed by flow cytometry. The osteogenic ability and adipogenic differentiation were confirmed with oil red O and alizarin red staining. The optimal concentration of bFGF to promote the proliferation of hUCMSCs was 20 μg/L. The cells in control group were cultured in the growth medium consisting of DMEM/F12 and 10% volume fraction of fetal bovine serum. The cells in experiment group were cultured under the same condition of control group but plus 20 μg/L bFGF. The proliferation of hUCMSCs was analyzed by MTT assay. The expression of type I and III collagens at mRNA and protein levels was determined by RT-PCR and Western blotting. RESULTS：The growth curves indicated that bFGF promoted the proliferation of hUCMSCs. The hUCMSCs expressed CD29, but did not express CD34, CD45 or HLA-DR in the presence or absence of bFGF unanimously. The cells were alizarin red staining-positive and oil red O staining-positive. Compared with control group, the expression of type I and III collagens significantly decreased at mRNA and protein levels in experiment group. CONCLUSION：bFGF promotes the proliferation of hUCMSCs and does not change the expression of the surface markers. bFGF inhibits the expression of type I and III collagens at mRNA and protein levels, indicating that bFGF enhances the healing of wound without inducing scar hyperplasia.     

Coriaria sinica Maxim extract promotes burn wound healing in rats

AIM：To investigate the effects of Coriaria sinica Maxim extract (CSME) on burn wound healing in rats. METHODS：Forty male SD rats were randomly divided into normal saline (NS) group, white petrolatum jelly (WPJ) group, silver sulfadiazine (SSD) group and CSME group. After the animals were anesthetized, the skin of their backs was burnt to induce deep Ⅱ degree burn wounds. These wounds were treated respectively for 21 d by covering dressings with NS, WPJ, SSD and CSME, respectively. On the 1st, 3rd, 7th, 14th and 21st days, after the clinical symptoms of the animals and conditions of the wounds such as the epithelization rate, crusting and hair growth were observed, the wound tissues were also taken for histological examination. The content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD), and the expression of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and collagen were detected. RESULTS：On the 21st day after burn, the new epithelial tissues in CSME group extraordinarily　developed and a great deal of hair also grew along the margin of the wounds. The epithelization rate of the wound tissues in CSME group was higher than that in other groups. There was rare new hair in the center of the wound area in SSD group, but intensive new hair was observed in CSME group. On the 14th day and 21st day after burn, multilayer of epithelial cells was entirely covered with the wound area in CSME group. Some of the healing signs, such as sufficient differentiation and a line up in order of collagen fibers, clear tissue structure, exceedingly active hyperplasia of sebaceous glands and hair follicles, and so on, were also observed in the wound area in CSME group. From the 1st day to 21st day after burn, the increase in protein expression of EGF and bFGF in the wound tissues was significantly higher than that in other groups in the early stage, which was quickly decreased and was obviously lower than other groups in the latter stage. The mRNA expression ratio of type I and III collagens in SSD group was significantly higher than that in other groups, while that in CSME group was significantly lower than that in other groups. CONCLUSION：CSME promotes burn wound healing without scarring. The effects of CSME are likely to associate with the expression reinforcement of EGF and bFGF at mRNA and protein levels in the early stage, and associate with the expression inhibition of them later. Moreover, CSME may also inhibit the mRNA expression of type I collagen and promote the synthesis of type III collagen in the wound tissues of burn.     

Effect of VEGF/bFGF complex peptide vaccine on toxicity and reproductivity in female-mice

AIM: To investigate the toxicity and reproductive effect of vascular endothelial growth factor (VEGF)/basic fibroblast growth factor (bFGF) complex peptide vaccine (VBP3) on the female-mice. METHODS: The VBP3 was purified with Ni-NTA affinity chromatography. The female BALB/c mice were immunized with the purified VBP3. The antibody titer in the serum was detected by ELISA. The data of the body weight and the organ weight of the parent mice were gathered and analyzed, and the pathological changes of the organs were observed with HE staining to investigate the toxicity of VBP3. To investigate the toxicity of VBP3 in the F1 mice, the parent immunized female mice were mated with the parent non-immunized male mice. After the F1 mice were born, the survival rate was calculated, the change of the body weight and the organ weight of the F1 mice were also determined. The pathological changes of the organs in F1 mice were also observed with HE staining. RESULTS: In the parent mice, high titers of the antibodies were detected against VEGF and bFGF, and no difference of the body weight, the organ weight and the pathological change between the immunized mice and control mice was found. In the F1 mice, a low titer of anti-bFGF antibody was detected compared with blank group. The survival rate in control group was higher than that in immunized group. In the 2 groups of the F1 mice, no obvious difference of the weight of the spleen, kidney, lung and heart was observed, and there was some difference in the weight of liver between the 2 groups. The results of the HE staining in the F1 mice showed some difference in the liver between the 2 groups. CONCLUSION: VEGF/bFGF complex peptide vaccine has no obvious organ toxicity in the parent female mice, but has some side effects on the reproductive and the healthy processes of F1 mice.     

The synergistic effect of endothelin-1 and basic fibroblast growth factor on rat vascular smooth muscle cell proliferation

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin-1 (ET-1) on rat aortic vascular smooth muscle cells (VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro-proliferative effect of VSMC. Western blotting was used to observe the variation of bFGF and FGFR-1 isoforms expression. RESULTS: bFGF and ET-1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose- and time-dependent. ET-1 increased all the three bFGF isoforms and FGFR-1 protein level in dose- and time-dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET-1 on bFGF and FGFR-1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET-1 had synergistic effect on VSMC proliferation. ET-1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR-1, which was PKC-dependent.     

Expression of bFGF in malignant tumor and its clinical pathological significance

AIM: To detect basic fibroblast growth factor (bFGF) expression in clinical common malignant tumor (non-small-cell lung cancer,breast cancer, colon cancer and melanoma), and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS: Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients (68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma).RESULTS: The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM. In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed. The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis. Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION: bFGF may participate in the process of occurrence and progression of malignant tumor. Expression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.     

双断根嫁接对辣椒嫁接苗愈合及生长的影响

以"巨根"辣椒为砧木,"龙椒六号"辣椒为接穗,以砧木不断根常规嫁接和辣椒自根苗分别作为对照,研究双断根套管嫁接方法对辣椒嫁接苗愈合及苗期生长的影响,以期为辣椒种植提供参考依据。结果表明:双断根套管嫁接可有效预防辣椒嫁接苗的徒长,显著提高壮苗指数。辣椒双断根套管嫁接时,砧木与接穗结合部的伤口愈合与砧木根系再生同时进行。嫁接可以有效提高根系活力,采取双断根套管嫁接后根系活力的提升更显著,根系活力随着嫁接时间的延长呈现降低趋势,双断根套管嫁接始终显著高于常规嫁接苗。2种嫁接方法之间的输导组织运输能力差异不显著;与自根苗相比,2种嫁接方法的输导能力在结合部愈合前期显著降低,在结合部完全愈合、嫁接苗成活后(嫁接后20 d)输导能力显著提高,且双断根方法略高于常规嫁接。     

Role of cAMP and cGMP in the regulation of bFGF gene expression of rat cardiomyocytes

AIM: To investigate the role of cAMP and cGMP in the regulation of bFGF gene expression of rat cardiomyocytes. METHODS: 8-Bromo-cAMP and sodium nitroprusside were added into the media to raise the concentrations of cAMP and cGMP of cultured cardiomyocytes, in situ hybridization and immunohistochemistry methods were used to identify the mRNA and protein expression of bFGF. The amount of bFGF mRNA and protein expression were detected by computor imaging analysis system MIAS exe. RESULTS: At 24 h after adding 8-Bromo-cAMP(0 1 mmol/L), bFGF mRNA and protein expression elevated significantly , at 48 h and 72 h it still expressed higher than that of control group( P< 0.05). But after adding sodium nitroprusside (1 mmol/L),bFGF expression decreased significantly,and especially at 48 h. CONCLUSION: The regulation of bFGF expression of cardiomyocytes is related to cAMP signal pathway.     

Adenosine promotes bFGF protein and bFGF mRNA expression in human umbilical vein endothelial cells in vitro

AIM: To investigate the influence of adenosine on human umbilical vein endothelial cells (HUVEC) bFGF protein production and bFGF mRNA expression. METHODS: Immunohistochemistry staining was performed to detect bFGF protein. RT-PCR was performed to detect bFGF mRNA expression. RESULTS: Immunohistochemistry study demonstrated that there was only a small amount of bFGF positive cells and the color was weak in control group (without adenosine). In groups treated with 10-4 mol/L and 10-6 mol/L adenosine, bFGF protein was significantly higher than that in control group (P<0.05). In 10-8 mol/L and 10-10 mol/L adenosine groups, there were no significant differences compared with control group (P>0.05). RT-PCR showed that in 10-4 mol/L and 10-6 mol/L adenosine groups, bFGF mRNA expression was higher than that in control group (P<0.05), while the difference between 10-8 mol/L adenosine group and control group was not significant (P>0.05). CONCLUSION: Adenosine may promote HUVEC proliferation and angiogenesis partly through inducing bFGF expression.     

Effects of bFGF and insulin on proliferation of mice cartilage tissue cells

AIM:To explore the effects of basic fibroblast growth factor ( bFGF) and insulin on the cell proliferation and differentiation in primary cartilage cells. METHODS:After induction with different concentrations of bFGF and insulin, cell proliferation was measured with WST-1 method and fluoroscope methods. RESULTS:bFGF and insulin exerted their related action on primary cartilage cells in 0.4% fatal bovine serum at different concentrations. 25 μg/L bFGF and 5 mg/L insulin promoted cell proliferation significantly. CONCLUSION:bFGF and insulin prolong the survival time and promote cell proliferation in primary cartilage cells.     

Expression of bovine bFGF-pcDNA3 eukaryotic vector in human umbilical vein endothelial cells and product identification

AIM: To study the expression and activity of bovine bFGF-pcDNA₃ in cultured human umbilical vein endothelial cells and its effect on the bioactivity of these cells. METHODS: Recombinant bovine bFGF-pcDNA₃ was transferred into cultured human umbilical vein endothelial cells by lipofectin transfection, and positive clones were selected by G418. The bFGF expression position and Ⅷ related antigen was examined by immunohistochemical analysis. Chemiluminescence Western blotting analysis of total cell extracts was carried to detect bFGF expression in transfected and non-transfected cells and to compare their expression level. Growth curves of transfected and non-transfected cells were drawn and doubling time were caculated. The activity of bFGF was estimated by MTT analysis. RESULTS: Human umbilical vein endothelial cells appeared elongated after transfection. Immunohistochemical analysis confirmed its endothelial origin both before and after transfection and demonstrated that both transfected and non-transfected cells express bFGF in cytoplasm. Western blotting confirmed that both cell lines express 17kD bFGF but the level is much higher in transfected cells. Growth curve of transfected cell line moves upward and the doubling time is shorter. MTT analysis confirmed that the bioactivity of the expression product is equal to about 571.4 ng/L of exogenous bFGF continuous stimulation. CONCLUSION: bFGF cDNA-transfected human umbilical vein endothelial cells can grow in vitro and express functional bFGF. bFGF-pcDNA₃ can increase endogenous bFGF expression and has corresponding bioactivity on human umbilical endothelial cells.     

Effect of bFGF on promoting angiogenesis in infarct area and improving myocardial fibrosis in mouse myocardial infarction model

AIM: To investigate the protective effect of basic fibroblast growth factor (bFGF) on the heart of mice with myocardial infarction and its mechanism. METHODS: The model of myocardial infarction was established by the ligation of left anterior descending artery of C57/B6 mice (8~12 weeks old) after lateral thoracotomy. The mice were divided into sham operation group, myocardial infarction group and bFGF administration group. bFGF at 0.5 μg was intraperitoneally injected on alternate days after myocardial infarction for 7 d. Cardiac Doppler ultrasonography was used to detect cardiac function after myocardial infarction for 28 d, and left ventricular end-diastolic diameter, left ventricular end-systolic diameter, left ventricular ejection fraction and left ventricular shortening fraction (LVFS) were used to evaluate cardiac function. After myocardial infarction for 28 d, the mice were sacrificed and the hearts were collected for preparing pathological sections. The degrees of myocardial fibrosis and angiogenesis in the myocardial infarction area were observed. Western blot was used to detect the indicators of angiogenesis. RESULTS: The results of Masson staining showed that bFGF administration significantly reduced myocardial fibrosis at 28 d after myocardial infarction. Cardiac ultrasound data showed that cardiac functions in myocardial infarction group were poorer than those in sham group, and bFGF administration significantly improved cardiac functions. Immunofluorescence staining showed that neovascularization in myocardial infarction area of bFGF administration group was more than that in myocardial infarction group. The results of Western blot showed that bFGF activated AKT/HIF-1α/VEGF signaling pathway. CONCLUSION: Intraperitoneal injection of bFGF reduces myocardial fibrosis and improves cardiac function in myocardial infarction mice. bFGF may promote angiogenesis by activating AKT/HIF-1α/VEGF signaling pathway.     

Inhibitory effect of different target-side antisense oligonucleotides on bFGF expression in SWO-38 cells

AIM:Three different antisense oligonucleotides complementary to basic fibroblast growth factor (bFGF) mRNA were compared in inhibitory effect on gene targeted expression.METHODS:After transfecting bFGF antisense oligonucleotides (asODN) into SWO-38 cells by lipofectin, the proliferation of cells was identified by MTT method, apoptosis was examined by flow cytometric cell cycle analysis and the expression levers of bFGF were detected by Western-blotting.RESULTS:There were 49%, 33%, 51% inhibition of cell growth and 35%, 27%, 18% cell apoptosis after asODN1, asODN2 and asODN3 treatment.In addition, the decrease in bFGF protein was 63%, 42%, 11%, respectively.CONCLUSION:The data suggeste that asODN1 is a potent target to bFGF mRNA, which inhibits cell growth and induces apoptosis in SWO-38 cells.     

Advances inthe research of bFGF binding to heparin

Basic fibroblast growthfactor (bFGF) is a multi-potential growth factor whose biological activities depend on its receptor’s intrinsic tyrosine kinase activity and second messengers such as the mitogen activated protein kinases. Heparin sulfate proteoglycans (HSPGs) have been demonstrated to enhance or inhibit bFGF activity. The response elicited by HSPGis related to the relative concentrations and binding kinetics for bFGF of the various pools of HSPG. The type of cellular response might depend on the specific HSPG and FGF receptor expressed on the cell surface. The specific core protein of HSPG, and tissue-specific differences in heparin sulfate modification result in altered bFGF regulation.     

Effect of basic fibroblast growth factor on synthesis of C-type natriuretic peptide in cultured human umbilical vein endothelial cells

AIM: To investigate the effect of basic fibroblast growth factor (bFGF) on C-type natriuretic peptide (CNP) production, release and mRNA expression. METHODS: Human endothelial cell cultured;CNP was mea sured by radioimmunoassay method;CNP mRNA expression was determined by RT-PCR technique.RESULTS: bFGF could augment CNP synthesis in human endothelial cells. Compared with control group,25 ng, 50 ng, 100 ng bFGF increased CNP contents in endothelial cells by 88% (P＜0.05), 95% (P＜0.05), 187% (P＜0.01), respectively.100 ng bFGF also stimulated CNP release from cultured human endothelial cell. In addition, 25 ng, 50 ng and 100 ng bFGF stimulated CNP mRNA expression of cultured human endothelial cells in a dose-dependent manner. CONCLUSION: bFGF might regulate CNP synthesis,release and mRNA expression in cultured umbilical human endothelial cells.     

Effects of probucol on the proliferation of rat vascular smooth muscle cells stimulated by basic fibroblast growth factor and hydrogen peroxide

AIM: To investigate the effect of probucol on proliferation of rat vascular smooth muscle cells(VSMC) stimulated by basic fibroblast growth factor (bFGF) and/or hydrogen peroxide(H₂O₂). METHODS: Effects of probucol on VSMC proliferation and DNA synthesis stimulated by bFGF and/or H₂O₂ were observed by means of MTT test, cell number count and [3H]-TdR incorporation. RESULTS: ①Probucol significantly inhibited proliferation and DNA synthesis in VSMC stimulated by bFGF and/or H₂O₂, with dosage-dependent manner. Cell number, A value and [3H]-TdR incorporation in group probucol+bFGF and group probucol+H₂O₂ were reduced by 40.0%, 39.1%, 45.5% and 46.9%, 45.0%, 39.5%, respectively, compared with group bFGF and group H₂O₂ (P＜0.05, P＜0.01, respectively). ②Pretreatment of VSMC with probucol for 24 h prior to bFGF and/or H₂O₂ stimulation exhibited significant inhibiton of VSMC proliferation and DNA synthesis, but after prestimulation by bFGF and/or H₂O₂ for 24 h, probucol had no influence on VSMC proliferation and DNA synthesis (P＞0.05). CONCLUSION: Probucol dramatically inhibited proliferation and DNA synthesis in VSMC stimulated by bFGF and/or H₂O₂, but had no inhibitory effect on the cell proliferation prestimulated by bFGF and /or H₂O₂.