Role of Daptomycin on Burn Wound Healing in an Animal Methicillin-Resistant Staphylococcus aureus Infection Model.

Prolonged hospitalization and antibiotic therapy are risk factors for the development of methicillin-resistant Staphylococcus aureus (MRSA) infections in thermal burn patients. We used a rat model to study the in vivo efficacy of daptomycin in the treatment of burn wound infections by S. aureus, and we evaluated the wound healing process through morphological and immunohistochemical analysis. A copper bar heated in boiling water was applied on a paraspinal site of each rat, resulting in two full-thickness burns. A small gauze was placed over each burn and inoculated with 5 × 107 CFU of S. aureus ATCC 43300. The study included two uninfected control groups with and without daptomycin treatment, an infected control group that did not receive any treatment, and two infected groups treated, respectively, with intraperitoneal daptomycin and teicoplanin. The main outcome measures were quantitative culture, histological evaluation of tissue repair, and immunohistochemical expression of wound healing markers: epidermal growth factor receptor (EGFR) and fibroblast growth factor 2 (FGF-2). The highest inhibition of infection was achieved in the group that received daptomycin, which reduced the bacterial load from 107 CFU/ml to about 103 CFU/g (P < 0.01). The groups treated with daptomycin showed better overall healing with epithelialization and significantly higher collagen scores than the other groups, and these findings were also confirmed by immunohistochemical data. In conclusion, our results support the hypothesis that daptomycin is an important modulator of wound repair by possibly reducing hypertrophic burn scar formation.

Blue light does not impair wound healing in vitro.

Irradiation with red or near infrared light promotes tissue repair, while treatment with blue light is known to be antimicrobial. Consequently, it is thought that infected wounds could benefit more from combined blue and red/infrared light therapy; but there is a concern that blue light may slow healing. We investigated the effect of blue 470nm light on wound healing, in terms of wound closure, total protein and collagen synthesis, growth factor and cytokines expression, in an in vitro scratch wound model. Human dermal fibroblasts were cultured for 48h until confluent. Then a linear scratch wound was created and irradiated with 3, 5, 10 or 55J/cm(2). Control plates were not irradiated. Following 24h of incubation, cells were fixed and stained for migration and fluorescence analyses and the supernatant collected for quantification of total protein, hydroxyproline, bFGF, IL-6 and IL-10. The results showed that wound closure was similar for groups treated with 3, 5 and 10J/cm(2), with a slight improvement with the 5J/cm(2) dose, and slower closure with 55J/cm(2) p<0.001). Total protein concentration increased after irradiation with 3, 5 and 10J/cm(2), reaching statistical significance at 5J/cm(2) compared to control (p<0.0001). However, hydroxyproline levels did not differ between groups. Similarly, bFGF and IL-10 concentrations did not differ between groups, but IL-6 concentration decreased progressively as fluence increased (p<0.0001). Fluorescence analysis showed viable cells regardless of irradiation fluence. We conclude that irradiation with blue light at low fluence does not impair in vitro wound healing. The significant decrease in IL-6 suggests that 470nm light is anti-inflammatory.

Combination Efficacy of Astragalus membranaceus and Curcuma wenyujin at Different Stages of Tumor Progression in an Imageable Orthotopic Nude Mouse Model of Metastatic Human Ovarian Cancer Expressing Red Fluorescent Protein.

BACKGROUND/AIM: The present study determined the efficacy of extracts of Astragalus membranaceus (AM) and Curcuma wenyujin (CW), a traditional Chinese medicine herbal mixture, at different tumor stages of an orthotopic nude mouse model of human ovarian cancer expressing red fluorescent protein. MATERIALS AND METHODS: The tumor-bearing mice were treated with cisplatinum (CDDP), AM, CW, or a combination of AM and CW in each of three tumor stages, using the same regimen. Group 1 received saline as negative control. Group 2 received CDDP i.p. as positive control with a dose of 2 mg/kg, every three days. Group 3 received AM daily via oral gavage, at a dose of 9120 mg/kg. Group 4 received CW daily via oral gavage, at a dose of 4560 mg/kg. Groups 5, 6 and 7 received combinations of AM and CW daily via oral gavage at low (AM, 2280 mg/kg; CW, 1140 mg/kg), medium (AM, 4560 mg/kg; CW 2280 mg/kg), and high (AM, 9120 mg/kg; CW, 4560 mg/kg) doses. The expression of angiogenesis- and apoptosis-related genes in the tumors were analyzed by immunohistochemistry for matrix metalloproteinase 2 (MMP-2), vascular endothelial growth factor (VEGF) fibroblast growth factor 2 (FGF-2), B-cell lymphoma 2 (Bcl-2) and cyclooxygenase 2 (Cox-2), and by polymerase chain reaction for MMP-2, FGF-2 and Bcl-2. RESULTS: CDDP, AM, and its combination with CW-induced significant growth inhibition of Stage I tumors. Strong efficacy of the combination of AM and CW at high dose was observed. Monotherapy with CDDP, AM, CW, and the combination treatments did not significantly inhibit Stage II and III tumors. The expression of MMP-2, VEGF, FGF-2, and Cox-2 was significantly reduced in Stage I tumors treated with AM, CW, and their combination, suggesting a possible role of these angiogenesis- and apoptosis-related genes in the observed efficacy of the agents tested. CONCLUSION: This study is the first report on the efficacy of anticancer agents at different stages of ovarian cancer in an orthotopic mouse model. As the tumor progressed, it became treatment-resistant, similar to the clinical situation, further demonstrating the utility of the model and the need for agents acrtive in advanced-stage ovarian cancer.

Restoration of impaired endothelial myocyte enhancer factor 2 function rescues pulmonary arterial hypertension.

BACKGROUND: Pulmonary arterial hypertension (PAH) is a progressive disease of the pulmonary arterioles, characterized by increased pulmonary arterial pressure and right ventricular failure. The cause of PAH is complex, but aberrant proliferation of the pulmonary artery endothelial cells (PAECs) and pulmonary artery smooth muscle cells is thought to play an important role in its pathogenesis. Understanding the mechanisms of transcriptional gene regulation involved in pulmonary vascular homeostasis can provide key insights into potential therapeutic strategies. METHODS AND RESULTS: We demonstrate that the activity of the transcription factor myocyte enhancer factor 2 (MEF2) is significantly impaired in the PAECs derived from subjects with PAH. We identified MEF2 as the key cis-acting factor that regulates expression of a number of transcriptional targets involved in pulmonary vascular homeostasis, including microRNAs 424 and 503, connexins 37, and 40, and Kruppel Like Factors 2 and 4, which were found to be significantly decreased in PAH PAECs. The impaired MEF2 activity in PAH PAECs was mediated by excess nuclear accumulation of 2 class IIa histone deacetylases (HDACs) that inhibit its function, namely HDAC4 and HDAC5. Selective, pharmacological inhibition of class IIa HDACs led to restoration of MEF2 activity in PAECs, as demonstrated by increased expression of its transcriptional targets, decreased cell migration and proliferation, and rescue of experimental pulmonary hypertension models. CONCLUSIONS: Our results demonstrate that strategies to augment MEF2 activity hold potential therapeutic value in PAH. Moreover, we identify selective HDAC IIa inhibition as a viable alternative approach to avoid the potential adverse effects of broad spectrum HDAC inhibition in PAH.

Role of sensory and motor intensity of electrical stimulation on fibroblastic growth factor-2 expression, inflammation, vascularization, and mechanical strength of full-thickness wounds.

Electrical stimulation (ES) profoundly affects angiogenesis by modulating the production of angiogenic factors. We evaluated the effect of sensory (direct current, 600 microamperes) and motor (monophasic pulse current, 2.5 to 3 milliamperes, 300-microsecond pulse duration, 100 Hz) intensities of cathodal current on the release of fibroblastic growth factor-2 (FGF-2) at the wound site and also the biomechanical and histological properties of healed skin. Ninety-six male, Sprague-Dawley rats were randomly assigned into one control and two experimental groups. A full-thickness skin incision was made on the dorsal region of each animal. The experimental groups received 10 sessions of ES (sensory or motor) for 1 hour per day every other day. The results showed that FGF-2 levels in the sensory group were significantly greater than in the other groups on the third day. In the motor group, FGF-2 levels were significantly decreased compared with the control group. There were no significant differences between the normalized ultimate strength and stiffness in the groups, but they tended to be higher in the motor ES group. We conclude that the application of sensory ES during the early stage of wound healing may have a beneficial effect on wound healing by inducing the release of angiogenic factors and decreasing the duration of the inflammation phase.

Low-level laser irradiation affects the release of basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), and receptor of IGF-I (IGFBP3) from osteoblasts.

OBJECTIVE: It was the aim of the present study to evaluate whether the laser irradiation of osteoblasts could enhance the release of growth factors including basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), and receptor of IGF-I (IGFBP3). BACKGROUND DATA: Low-level laser therapy (LLLT) has been shown to have biostimulatory effects on various cell types by enhancing production of some cytokines and growth factors. MATERIALS AND METHODS: Human mesenchymal stem cells (MSCs) were seeded in osteogenic medium and differentiated into osteoblasts. Three groups were formed: in the first group (single dose group), osteoblasts were irradiated with laser (685 nm, 25 mW, 14.3 mW/cm(2), 140 sec, 2 J/cm(2)) for one time; and in the second group, energy at the same dose was applied for 2 consecutive days (double dose group). The third group was not irradiated with laser and served as the control group. Proliferation, viability, bFGF, IGF-I, and IGFBP3 levels were compared between groups. RESULTS: Both of the irradiated groups revealed higher proliferation, viability, bFGF, IGF-I, and IGFBP3 expressions than did the nonirradiated control group. There was increase in bFGF and IGF-I expressions and decrease in IGFBP3 in the double dose group compared to single dose group. CONCLUSIONS: The results of the present study indicate that LLLT increases the proliferation of osteoblast cells and stimulates the release of bFGF, IGF-I, and IGFBP3 from these cells. The biostimulatory effect of LLLT may be related to the enhanced production of the growth factors.

Effect of laser phototherapy on the release of fibroblast growth factors by human gingival fibroblasts.

The effects of laser phototherapy on the release of growth factors by human gingival fibroblasts were studied in vitro. Cells from a primary culture were irradiated twice (6 h interval), with continuous diode laser [gallium-aluminum-arsenium (GaAlAs), 780 nm, or indium-gallium-aluminum-phosphide (InGaAlP),_660 nm] in punctual and contact mode, 40 mW, spot size 0.042 cm(2), 3 J/cm(2) and 5 J/cm(2) (3 s and 5 s, respectively). Positive [10% fetal bovine serum (FBS)] and negative (1%FBS) controls were not irradiated. Production of keratinocyte growth factor (KGF) and basic fibroblast growth factor (bFGF) was quantified by enzyme-linked immunosorbent assay (ELISA). The data were statistically compared by analysis of variance (ANOVA) followed by Tukey's test (P

Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from gingival fibroblasts.

Various studies have shown biostimulation effects of laser irradiation by producing metabolic changes within the cells. Little is known about the biological effect of laser irradiation on the oral tissues. Among the many physiological effects, it is important to recognize that low-level laser therapy (LLLT) may affect release of growth factors from fibroblasts. Therefore, the aim of the present study was to determine whether the laser irradiation can enhance the release of basic fibroblast growth factor (bFGF), insulin-like growth factor-1 (IGF-1), and receptor of IGF-1 (IGFBP3) from human gingival fibroblasts (HGF). The number of all samples in the study were 30, and the samples were randomly divided into three equal groups; In the first group (single dose group), HGF were irradiated with laser energy of 685 nm, for 140 s, 2 J/cm(2) for one time, and in the second group, energy at the same dose was applied for two consecutive days (double dose group). The third group served as nonirradiated control group. Proliferation, viability, and bFGF, IGF-1, IGFBP3 analysis of control and irradiated cultures were compared with each other. Both of the irradiated groups revealed higher proliferation and viability in comparison to the control group. Comparison of the single-dose group with the control group revealed statistically significant increases in bFGF (p < 0.01) and IGF-1 (p < 0.01), but IGFBP3 increased insignificantly (p > 0.05). When the double dose group was compared with the control group, significant increases were determined in all of the parameters (p < 0.01). In the comparison of the differences between the two irradiated groups (one dose and two doses), none of the parameters displayed any statistically significant difference (p > 0.05). In both of the laser groups, LLLT increased the cell proliferation and cell viability. The results of this study showed that LLLT increased the proliferation of HGF cells and release of bFGF, IGF-1, and IGFBP3 from these cells. LLLT may play an important role in periodontal wound healing and regeneration by enhancing the production of the growth factors.

[Effect of panax quinquefolius saponin on angiogenesis and expressions of VEGF and bFGF in myocardium of rats with acute myocardial infarction].

OBJECTIVE: To study the effect of Panax quinquefolius Saponin (PQS), an extraction from stem and leaf of American ginseng, on vascular regeneration in infarcted area, and expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in myocardium of rats with acute myocardial infarction (AMI). METHODS: Fifty male Wistar rats were established into AMI model successfully and randomly divided into 5 groups equally, i.e. the model group, the high, middle and low dose PQS groups and the metoprolol group. They were treated with saline, different doses of PQS (54 mg/kg x d, 27 mg/kg x d, 13.5 mg/kg x d)and metoprolol (4.5 mg/kg x d) respectively, by gavage once a day for 14 days. Besides, a sham operated group and a normal control group were set up for control with 10 rats in each group. All rats were killed on the 15th day. Six samples of heart were chosen from each group for examining expressions of VEGF, bFGF and the VIII coagulation factor under light microscopy by immunohistochemical staining, and the quantitative analysis on positive responsive intensity of VEGF and bFGF was conducted on the other 4 heart samples using the image analysing system, then mean micro-vessel density (MMVD) was calculated. RESULTS: The expressions of VEGF and MMVD were higher in the high and the middle dose PQS groups than those in the model group (P < 0.05) and the expression of bFGF was higher in the three PQS groups than that in the model group (P < 0.05). CONCLUSION: PQS can protect myocardium from ischemic injury in rats after AMI by way of promoting angiogenesis in the infarcted or ischemic area of myocardium and up-regulating expressions of VEGF and bFGF in myocardial cells.

Magnolia ovovata extract and its active component magnolol prevent skin photoaging via inhibition of nuclear factor kappaB.

Transcriptional activity of nuclear factor kappaB (NF-kappaB) is induced by environmental signals including inflammation, UV irradiation and oxidative stress. It was shown that the NF-kappaB activity greatly contributes to the skin photoaging process. Thus, it is plausible that NF-kappaB inhibitors could directly prevent skin photoaging. In this study, we found that Magnolia ovovata extract inhibited NF-kappaB-mediated gene expression and demonstrated that external swabbing with Magnolia extract preventing skin photoaging processes through keratinocyte hyperproliferation and degradation of collagen fibers in mice skin. We have identified magnolol as the solely responsible active compound in Magnolia extract. Magnolol effectively inhibited the NF-kappaB-dependent transcription, but no effect was observed with other inducible transcription factors such as activator protein-1 (AP-1) and cyclic-AMP responsive element-binding protein (CREB). In addition, magnolol was effective in inhibiting the production of basic fibroblast growth factor (bFGF) and matrix metalloprotease-1 (MMP-1) from the cells overexpressing p65, a major subunit of NF-kappaB. Although magnolol did not affect the phosphorylation and degradation of IkappaBalpha, it inhibited the nuclear translocation of the activated NF-kappaB. These findings suggest that Magnolia extract and its active component magnolol can be used to prevent the skin photoaging via inhibiting NF-kappaB by external topical application.

Influence of Echinacea purpurea intake during pregnancy on fetal growth and tissue angiogenic activity.

The process of angiogenesis and control of blood vessels sprouting are fundamental to human health, as they play key roles in many physiological and pathological conditions. Intake of different pharmaceuticals with antiangiogenic activity by pregnant women may lead to severe developmental disturbances as it was described in case of thalidomide. It may also cause immunomodulatory effects as it was shown for antibiotics, theobromine, caffeic acid or catechins on the pregnant mice model. At present, Echinacea purpurea-based phytoceuticals are among the most popular herbals in the marketplace. Many compounds of Echinacea extracts (polysaccharides, alkamides, polyphenols, glycoproteins) exert immunomodulatory, anti-oxidative and anti-inflammatory activity. Echinacea is one of the most powerful and effective remedies against many kinds of bacterial and viral infections. In previous studies we shown significant inhibitory effect of the Echinacea purpurea based remedy on tumour angiogenic activity using cutaneous angiogenesis test, and an inhibitory effect on L-1 sarcoma growth was observed . The aim of the present study was to establish whether pharmaceuticals containing alcoholic extracts of Echinacea purpurea given to pregnant mice influence angiogenic activity and tissue VEGF and bFGF production of their fetuses. We showed that angiogenic activity of tissue homogenates was increased in Esberitox group and diminished in case of Immunal forte as compared to standard diet group. In case of Echinapur group we did not find significant differences in angiogenic activity. VEGF and bFGF concentration were lower in all groups compared to the control. In the case of Echinapur and Esberitox number of fetuses in one litter were slightly lower as compared to control group, but the difference is on the border of statistical significance. In conclusion, there is some possibility that pharmaceuticals containing Echinacea purpurea might influence fetal development in human also, because they may interfere with embrional angiogenesis , and should not be recommended for pregnant women.

Combination therapy of human pancreatic cancer implanted in nude mice by oral fluoropyrimidine anticancer agent (S-1) with interferon-alpha.

PURPOSE: We evaluated the antitumor and antiangiogenic activities of human natural interferon-alpha (IFN-alpha) alone or in combination with S-1 against human pancreatic cancer cells. METHODS: Three days after the subcutaneous (s.c.) implantation of tumor cells, mice (n = 12) were received s.c. injection with IFN-alpha alone (10,000 U six times a week), oral administration with S-1 alone (8 mg/kg six times a week), or both with IFN-alpha and S-1 (8, 10, 12 mg/kg six times a week). RESULTS: Administration of IFN-alpha in combination with S-1 significantly decreased progressive growth and angiogenesis of human pancreatic cancer cells. The combination therapy produced more significant inhibition in expression of the representative proangiogenic molecules, vascular endothelial growth factor and basic fibroblast growth factor than individual treatment either IFN-alpha or S-1 alone did. These treatments also decreased the staining of proliferating cell nuclear antigen, induced apoptosis and decreased microvessel density. In order to better understand the precise molecular mechanisms by which IFN-alpha and S-1 exert its effects, we have utilized cDNA microarray including 124 known genes to determine the gene expression profile altered by IFN-alpha and S-1 treatment. We found a total of seven genes which showed a twofold change after IFN-alpha and S-1 treatment in addition to VEGF, bFGF, CD31, MMP-2, MMP-7 and MMP-9. Among these genes, we found down-regulation of six genes and up-regulation of one gene, which are related to angiogenesis, tumor cell invasion and metastasis. CONCLUSIONS: These data suggest that administration of IFN-alpha in combination with S-1 may provide a novel and effective approach to the treatment of human pancreatic cancer.

[Protective and reverse effects of qindan capsule on aortic lesion in spontaneously hypertensive rats].

OBJECTIVE: To investigate the effects and mechanism of Qindan Capsule (QC) on aortic structure in spontaneously hypertensive rats (SHR). METHODS: Thirty-two SHR of 14 weeks old, were divided into the QC group, the Niuhuang Jiangya Capsule (NJC) group, the captopril group and the model group. Besides, Wistar-Kyoto (WKY) rats were taken as the normal control. All the others were administered with corresponding medicine and their blood pressure measured. After 12 weeks, the morphological changes of aorta were observed by HE and Masson staining, the level of angiotensin II (Ang II) in aorta was detected by radioimmunoassay, and the mRNA expression of basic fibroblast growth factor (bFGF) in aortic wall was analyzed by real-time quantitative fluorescent PCR. RESULTS: QC could reduce the blood pressure in SHR, improve their aortic structure, lower the Ang II level and inhibit the bFGF mRNA expression in aortic wall (P < 0.05 or P < 0.01), showing a good effect similar to that of captopril (P > 0.05) and better than that of NJC (P < 0.01). CONCLUSION: QC has a significant protective and reverse effect on aortic lesion in SHR. The mechanism may be related to its actions in reducing Ang II level and inhibiting bFGF mRNA expresion in aortic wall.

High-dose atorvastatin improves hypercholesterolemic coronary endothelial dysfunction without improving the angiogenic response.

BACKGROUND: Although 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) can restore endothelial function in coronary disease, in vitro and murine studies have shown their effects on myocardial angiogenesis to be biphasic and dose dependent. We investigated the functional and molecular effects of high-dose atorvastatin on the endogenous angiogenic response to chronic myocardial ischemia in hypercholesterolemic swine. METHODS AND RESULTS: Yucatan pigs were fed either a normal (NORM group; n=7) or high-cholesterol diet, with (CHOL-ATR group; n=7) or without (CHOL group; n=6) atorvastatin (3 mg/kg per day) for 13 weeks. Chronic ischemia was induced by ameroid constrictor placement around the circumflex artery. Seven weeks later, microvessel relaxation responses, myocardial perfusion, and myocardial protein expression were assessed. The CHOL group demonstrated impaired microvessel relaxation to adenosine diphosphate (29+/-3% versus 61+/-6%, CHOL versus NORM; P<0.05), which was normalized in the CHOL-ATR group (67+/-2%; P=NS versus NORM). Collateral-dependent myocardial perfusion, adjusted for baseline, was significantly reduced in the CHOL group (-0.27+/-0.07 mL/min per gram versus NORM; P<0.001) as well as the CHOL-ATR group (-0.35+/-0.07 mL/min per gram versus NORM; P<0.001). Atorvastatin treatment was associated with increased phosphorylation of Akt (5.7-fold increase versus NORM; P=0.001), decreased vascular endothelial growth factor expression (-68+/-8%; P<0.001 versus NORM), and increased expression of the antiangiogenic protein endostatin (210+/-48%; P=0.004 versus NORM). CONCLUSIONS: Atorvastatin improves hypercholesterolemia-induced endothelial dysfunction without appreciable changes in collateral-dependent perfusion. Increased myocardial expression of endostatin, decreased expression of vascular endothelial growth factor, and chronic Akt activation associated with atorvastatin treatment may account for the diminished angiogenic response.

[Arnebia root oil promotes histological change and up-regulates bFGF and it's mRNA expression in the raw surface of the rabbits].

OBJECTIVE: To explore the molecular biological mechanism of arnebia root oil in promoting wound surface healing by observing histological change and basic fibroblast growth factor(bFGF) mRNA expression in the wound surface tissues of 2 groups, as well as the wound surface healing rate. METHOD: Experimental model of incised-wound was produced on the back of 18 New Zealand albino rabbits. The wound surfaces were randomly divided into two groups, namely, experimental group and control group. The wound surfaces in the experimental group were treated by arnebia root oil and those in control group were treated by petrolatum gauze. Then raw surfaces were evaluated by the techniques of histology, histochemistry and electron microscope and the healing rates of the raw surfaces were compared between the two groups. Content of bFGF and it's mRNA expression in wound surface tissue was also measured by means of Western-blot and RT-PCR. RESULT: The wound surface healing rate in experimental group was higher than that in control group( P < 0.05). The fibroblast, collagen and blood capillaries were comparatively richer in experimental group as compared with those in control group, and similarly, the expression of bFGF mRNA was also significantly enhanced in the experimental group as compared with control group during the various periods of treatment. In addition, the changes in the expressions of bFGF and it's mRNA paralleled the changes of healing rates in the two groups. CONCLUSION: the present results showed that amebia root oil significantly can promote the healing of raw surfaces, which may be mediated by up-regulation of bFGF expression.

[Inhibitory effects of ampelopsin on angiogenesis].

OBJECTIVE: To study the effect of ampelopsin on angiogenesis. METHODS: The anti-angiogenic effect was evaluated by MTT assay for proliferation of endothelial cells. The concentration of vascular endothelial growth factor (VEGF) and basic-fibroblast growth factor (bFGF) from human hepatocellular carcinoma Bel-7402 cells were detected by enzyme linked immunosorbant assay (ELISA). Immunohistochemical staining was conducted to detect the expression of VEGF and bFGF. The VEGF and bFGF in the cancer cells were examined by flow cytometry. The inhibitory effect of ampelopsin on the growth of human hepatocellular carcinoma Bel-7402 in nude mice was studied. RESULTS: Ampelopsin was shown to inhibit the proliferation of primary cultured bovine aortic endothelial cells in a concentration dependent manner in range of 6.4 - 51.2 microg/ml. The IC50 (50% inhibition concentration) value was 22.0 +/- 4.0 microg/ml. ELISA assay was shown that treatment with 12.8 microl/m1, 25.6 microl/ml and 38.4 microg/ml of ampelopsin resulted in an inhibition of VEGF production released by Bel-7402, and the inbibtitory rate was 14.2%, 40.0% and 49.6%, respectively. After exposure to 12.8 microg/ml of ampelopsin, a decrease in the expression and activity of VEGF and bFGF was observed by immunohistochemical staining. The concentration of VEGF and bFGF secretion by Bel-7402 cells were lower following ampelopsin treatment as shown by flow cytometry. Treatment with 25.6 microg/mL and 38.4 microg/ml of ampelopsin, the inbibitory rates were 32.2% and 57.4% for VEGF, and 54.9% and 62.6% for bFGF, respectively. The inhibitory rate of ampelopsin to the growth of the transplant tumor in nude mice were 24.3%, 41.4% and 45.75 respectively at the dose of 100 mg/kg, 150 mg/kg and 200 mg/kg. CONCLUSION: Ampelopsin is a potent inhibitor of VEGF and bFGF expression and production in human hepatocellular carcinoma Bel-7402 cell, and may be a promising angiogenesis inhibitor.

[Arnebia root oil promotes wound healing and expression of basic fibroblast growth factor on the wound surface in rabbits].

OBJECTIVE: To observe the promoting effect of arnebia root oils on expression of basic fibroblast growth factor (bFGF) in skin wound of rabbits and the histomorphological changes in the wound surface, and to discuss its mechanism. METHODS: Bilateral round skin wounds were made on the back of 15 rabbits. The three wounds on one side of the back of each rabbit were treated with arnebia root oils, while the three wounds on the other side were treated with vaseline in order to promote the wound healing. The histomorphology and ultrastructure under electron microscopy of the wounds, and the rate of wound healing were examined at different time. Western blotting assay was used to detect the expression of bFGF in the wound surface. RESULTS: The healing rate of the arnebia root oils-treated wounds was evidently higher than that of the vaseline-treated wounds (P<0.05). The quantities of fibroblast, collagen and capillary in the arnebia root oils-treated wounds were much more than those in the vaseline-treated wounds, and the expression of endogenous bFGF in the arnebia root oils-treated wounds was enhanced obviously as compared with that in the vaseline-treated wounds in different period of wound healing. There existed a parallel correlation between the expression level of bFGF and the rate of wound healing. CONCLUSION: The promoting effect of arnebia root oils on wound healing may be related to increasing the expression level of basic fibroblast growth factor in the skin wound.

[Effect of arnebia root oil in promoting the tissue recovery of surface of wound and basic fibroblast growth factor (bFGF) mRNA expression].

OBJECTIVE: To explore the molecular biological mechanism of Arnebia Root oil (AO) in promoting the recovery of surface of wound by observing basic fibroblast growth factor (bFGF) mRNA expression in the wound tissue and healing rate of the wound. METHODS: Patients in the observed group with their wound treated by AO and those in the control group treated by petrolatum gauze. The wound surface healing rate was estimated and bFGF mRNA expression was observed by RT-PCR. RESULTS: Endogenous bFGF mRNA expression existed in the wound surface of both groups, but its level in the observed group at any time point was obviously higher than that in the control group respectively, with significant difference in comparison of the gray density between the two groups (P < 0.05). The wound surface healing rate kept abreast with bFGF mRNA expression in wound tissues, so it was significantly higher in the observed group than that in the control group (P < 0.05). GAPDH gene, which was taken as a parameter for internal reference, expressed with a certain amount unchanged in different periods of healing (P > 0.05 ). CONCLUSION: AO shows obviously promotive action on bFGF, an important regulatory factor on wound healing, it might complete the recovery process by stimulating the increase of bFGF.

Combined effect of hyberbaric oxygen and N-acetylcysteine on fibroblast proliferation.

OBJECTIVE: To examine the combined effect of hyperbaric oxygen and N-acetylcysteine, a well-studied antioxidant, on fibroblast proliferation and production of 3 specific growth factors: basic fibroblast growth factor, vascular endothelial growth factor, and transforming growth factor beta1. DESIGN: In vitro study. SUBJECTS: None. INTERVENTIONS: Human dermal fibroblasts were propagated in serum-free medium and subjected to daily 90-minute 2-atm hyperbaric oxygen treatments with varying concentrations of N-acetylcysteine for 7 consecutive days. Cell proliferation and growth factor assays were performed on days 0, 1, 3, 5, and 7. RESULTS: Population doubling time decreased significantly with 40 micromol/L of N-acetylcysteine supplementation of 2-atm hyperbaric oxygen treatment. Higher levels of N-acetylcysteine increased population doubling time. CONCLUSIONS: Supplementation of hyperbaric oxygen therapy with 40 mumol/L of N-acetylcysteine appears to increase fibroblast proliferation without producing an unfavorable growth factor profile for normal healing. This suggests that this level of N-acetylcysteine may foster an ideal redox environment for fibroblast proliferation in a hyperbaric oxygen environment.

Differential effects of reproductive and hormonal state on basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in the hypothalamus and cingulate cortex of female rats.

Morphological changes in astrocytes occur in a number of brain regions including the hypothalamus and hippocampal regions as a function of hormonal and reproductive state. Because basic fibroblast growth factor has been shown to play an important role in morphological changes in astrocytes, we investigated whether basic fibroblast growth factor immunoreactivity would also be influenced by reproductive state and circulating gonadal steroids. To do this we compared astrocytic basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in hypothalamic nuclei and the cingulate cortex, area 2 among groups of cycling, late pregnant and lactating rats as well as in ovariectomized and ovariectomized hormone-replaced females. Significant differences in both basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity were observed across groups in the supraoptic nucleus, parvocellular paraventricular nucleus, medial preoptic area of the hypothalamus and cingulate cortex 2. The pattern of change in basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity varied across regions both in direction and magnitude. For example, although in the supraoptic nucleus ovariectomized rats had lower levels of basic fibroblast growth factor-ir than cycling females, this pattern was reversed within cingulate cortex. Overall the results of this study suggest that reproductive and hormonal states are associated with robust changes in basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in a number of brain areas but that the changes observed vary in magnitude as well as direction from one brain region to another.