A number of structural changes occur in the airway wall in asthma. The most characteristic is thickening of the subepithelial lamina reticularis which is observed in bronchial tissue even in patients with mild disease. This pathophysiological change which was the result of deposition of interstitial collagens by increased numbers of myofibroblasts is likely to be directed by growth factors having fibropoliferative and profibrotic effect. The acivation of the epithelial-mesenchymal unit involves reciprocal activities of growth factors belonging to the fibroblast growth (FGF), epidermal growth factor (EGF), and transforming growth factor-beta families. Among them FGF is a member of family of heparin binding growth factors that affect the growth and differentiation of a large number of cell types. Especially basic FGF involved in morphogenesis, wound repair, inflammation, angiogenesis, and tumor growth and invasion, and require the glycosaminoglycan side chains of heparan sulphate, proteoglycans for high affinity binding to their specific receptors. Few studies suggested bFGF would be an important regulator of airway remodeling by means of paracrine control of bronchial myofibroblasts in response to cell damage and repair.
Airway Remodeling* ; Asthma* ; Collagen ; Epidermal Growth Factor ; Fibroblast Growth Factors ; Fibroblasts ; Heparin ; Humans ; Inflammation ; Intercellular Signaling Peptides and Proteins ; Morphogenesis ; Myofibroblasts ; Proteoglycans ; Wounds and Injuries

BACKGROUND: A loss of bone mass is usually detected after a bone marrow transplantation (BMT), especially during the early post-transplant period. We recently reported that enhanced bone resorption following a BMT was related to both the steroid dose and the increase in IL-6. We also suggested damage to the marrow stromal microenvironment, by myoablation, partly explains the impaired bone formation following a BMT. It is well known that some growth factors play important role in bone growth and osteogenesis. However, the pathogenetic role of bone growth factors in post-BMT bone loss is unknown and data on the changes in the growth factors, in accordance with bone turnover markers and bone mineral density (BMD) changes are scarce. We investigated changes in bone growth factors such as IGF-I (Insulin-like growth factor-I), fibroblast growth factor-2 (FGF-2) and Macrophage colony stimulating factor (M-CSF), during the post-BMT period, and assessed whether the growth factor changes influenced the bone turnover and post-BMT bone loss. The present study is the first prospective study to describe the changes in bone growth factors following a BMT. METHODS: We prospectively investigated 110 patients undergoing a BMT, and analyzed 36 patients (32.4+/-1.3 years, 17 men and 19 women) whose BMDs were measured before, and 1 year after, the BMT. The serum biochemical markers of bone turnover were measured before, 1, 2, 3 and 4 weeks, 3 and 6 months, and 1 year, after the BMT. The serum FGF-2, IGF-I and M-CSF levels were measured before and 1 and 3 weeks, and 3 months after the BMT. The correlation between the changes of growth factors and various bone parameters was analyzed. RESULTS: The mean bone losses in the lumbar spine and total proximal femur, calculated as the percentage change from the baseline to the level at 1 year, were 5.2 (p<0.05) and 11.6% (p<0.01), respectively. The serum type I carboxyterminal telopeptide (ICTP), a bone resorption marker, increased progressively until 6 months after the BMT, but thereafter decreased, to the base value after 1 year. Serum osteocalcin, a bone formation marker, decreased progressively, until 3 weeks after the BMT but then increased transiently, and finally returned to the base level at 1 year. The serum IGF-I and FGF-2 also decreased progressively until 3 weeks and 1 week after the BMT, respectively, then increased to the base values at 3 months. The serum M-CSF increased briskly at 1 week post-BMT, then decreased to the base level. There were positive correlations between the percentage changes from the baseline proximal femur BMD and the IGF-I levels 3 weeks and 3 months (r=0.52, p<0.01, r=0.41, p<0.05) post BMT. A Significant correlation was found between the IGF-I and osteocalcin levels pre-BMT, and 3 weeks after the BMT. Another positive correlation was found between the M-CSF and the ICTP levels at 3 weeks post BMT (r=0.54, p<0.05). CONCLUSION: In conclusion, there were significant changes in the serum IGF-I, FGF-2 and M-CSF levels in the immediate post-BMT period, which were related to a decrease in bone formation and loss in the proximal femoral BMD during the year following the BMT
Biomarkers ; Bone Density ; Bone Development ; Bone Marrow ; Bone Marrow Transplantation ; Bone Resorption ; Colony-Stimulating Factors ; Femur ; Fibroblast Growth Factor 2 ; Hematopoietic Stem Cell Transplantation* ; Hematopoietic Stem Cells* ; Humans ; Insulin-Like Growth Factor I ; Intercellular Signaling Peptides and Proteins* ; Interleukin-6 ; Macrophage Colony-Stimulating Factor ; Macrophages ; Male ; Metabolism* ; Osteocalcin ; Osteogenesis ; Osteoporosis ; Prospective Studies ; Spine

PURPOSE: The objective of this study was to characterize transdifferentiated lens epithelial cells analyzed by reverse transcription-polymerase chain reaction (RT-PCR) for the expression of mRNAs encoding growth factors, growth factor receptors and pathologic extracellular matrix proteins and by Western blot analysis for the proteins encoded by these mRNAs. Moreover, after antioxidants treatment, such as Nacetyl cysteine (NAC), we observed the effect on changes in the expression of growth factors, growth factor receptors and extracellular matrix proteins. METHODS: TGF-beta treated rat lens cultured with medium 199 (Sigma Co. St. Louis, MO) was subject to RT-PCR and Western blot analysis to assess expression of mRNAs and proteins encoded by these mRNAs. RESULTS: The expression of mRNAs for TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta receptor, epidermal growth factor (EGF), epidermal growth factor receptor, fibroblast growth factor (FGF), fibroblast growth factor receptor and connective tissue growth factor (CTGF) were increased. The levels of type I collagen, fibronectin, and alpha-smooth muscle actin (SMA) mRNAs were also increased. However, the expression of growth factors, receptors, extracellular matrix were decreased by antioxidant, such as NAC. CONCLUSIONS: The enhanced expression of growth factors, growth factor receptors and extracellular matrix in present the molecular mechanism underlying pathogenesis of cataracts. And the suppression of growth factors and growth factor receptors with treatment of antioxidants, such as NAC, suggests the possibility of using drugs in the prevention or treatment of cataracts.
Actins ; Animals ; Antioxidants ; Blotting, Western ; Cataract ; Collagen Type I ; Connective Tissue Growth Factor ; Cysteine ; Epidermal Growth Factor ; Epithelial Cells ; Extracellular Matrix Proteins ; Extracellular Matrix* ; Fibroblast Growth Factors ; Fibronectins ; Intercellular Signaling Peptides and Proteins ; Rats* ; Receptor, Epidermal Growth Factor ; Receptors, Fibroblast Growth Factor ; Receptors, Growth Factor ; Receptors, Transforming Growth Factor beta ; RNA, Messenger ; Transforming Growth Factor beta

OBJECTIVES: To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism. METHODS: Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot. RESULTS: Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05). CONCLUSIONS Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.
Animals ; CCN Intercellular Signaling Proteins ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glucosamine ; pharmacology ; Intercellular Signaling Peptides and Proteins ; metabolism ; Osteoarthritis, Knee ; drug therapy ; Proto-Oncogene Proteins ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Wnt Signaling Pathway ; Wnt1 Protein ; metabolism ; beta Catenin ; metabolism

To investigate the effects of fibronectin (FN), hyaluronic acid (HA), epidermal growth factor (EGF), fibroblast growth factor (FGF), and transforming growth factor-beta(TGF-beta) on the healing of the corneal endothelial cells, the rabbit corneal endothelial cells were cultured from the outgrowth of Descemet's membrane explants. After making a wound at the center of the monolayer of confluent corneal endothelial cells using a rotating silicone tip, three different concentrations of testing agents were added. Then, the wound healing area and healing rate were measured. In addition, the effects of agents on the DNA synthesis by corneal endothelial cells were determined by measuring the incorporation of 3H-thymidine into cold acid-insoluble material. FN and EGF increased the wound healing area at all concentrations tested, and HA only at 1.0mg/ml showed a stimulatory effect. However, FGF showed little effect at all concentrations. TGF-beta promoted wound healing at 1.0 and 10.0 ng/ml. The combined treatment of FN and HA caused a more potent effect on corneal endothelial wound healing than single treatment. Neither the combined treatment of EGF and TGF-beta, nor the combined treatment of FGF and TGF-beta caused any significant effect. Wound healing rate was highest during the first 12 hours in all agents. EGF stimulated DNA synthesis while other agents studied did not, and TGF-beta suppressed EGF-induced DNA synthesis. These results indicate that FN, HA and TGF-beta stimulate the endothelial wound healing by probably enhancing cell migration, and EGF by enhancing cell migration and cell proliferation.
Cell Movement ; Cell Proliferation ; Descemet Membrane ; DNA ; Endothelial Cells* ; Epidermal Growth Factor ; Fibroblast Growth Factors ; Fibronectins* ; Hyaluronic Acid* ; Intercellular Signaling Peptides and Proteins* ; Silicones ; Transforming Growth Factor beta ; Wound Healing* ; Wounds and Injuries*

PURPOSE: To evaluate the effect of mitomycin-C (MMC) on osteotomy site as an adjunctive therapy for dacryocystorhinostomy, the effect of MMC on cultured human osteoblasts was tested. METHODS: Cultured osteoblasts which was obtained from the human iliac crest, were treated with four different concentrations of MMC (0 mg/ml, 0.2 mg/ml, 0.02 mg/ml, 0.002 mg/ml) and cultured for 24hours. To observe the effect of exposed time dependency, cells were treated with MMC during 5, 30minutes, and 24hours and washed and changed with fresh osteogenic media, and then cultured for 24 hours. The effect of fibroblast growth factor (FGF) and transforming growth factor-beta(TGF-beta) on the MMC-treated cells was evaluated. Cell viability was measured using trypan blue staining method and MTT assay. RESULTS: As compared with control group, the lowest growth rate of osteoblasts was 6.8% in 0.2 mg/ml MMC-treated cells. There were no significant differences in the growth rate between 5 minutes and 30 minutes MMC treatment groups, but in case of 24 hours treatment group with MMC (0.2 mg/ml) the growth rate was suppressed to 77.5% of control group with statistical significance. Both growth factors had promotive effect on the growth of in 0.02 mg/ml MMC-treated osteoblasts, but not in 0.2 mg/ml MMC-treated cells. CONCLUSIONS: Osteoblasts which were treated for longer time and with higher concentration of MMC showed more suppression in growth rate. These results suggest that intraoperative application of MMC during dacryocystirhinostomy could have a positive effect of mucosal ostium with suppression of osteoblasts proliferation.
Cell Survival ; Dacryocystorhinostomy ; Fibroblast Growth Factors ; Humans* ; Intercellular Signaling Peptides and Proteins ; Mitomycin* ; Osteoblasts* ; Osteotomy ; Trypan Blue

PURPOSE: To evaluate the effect of mitomycin C (MMC) on osteotomy site as an adjunctive therapeutic agent during dacryocystorhinostomy, the effect of MMC on cultured rabbit osteoblasts was tested. METHODS: Cultured osteoblasts which was obtained from the iliac crest of rabbits, were treated with MMC (0.2 mg/ml) for 5 or 30 minutes, washed and changed with fresh osteogenic media (Opti-MEM), and then cultured for 24 hours. To observe the effect of MMC dose dependency on cultured osteoblasts, four different concentrations of MMC (0.2 mg/ml, 0.02 mg/ml, 0.002 mg/ml, 0.0002 mg/ml) were applied into the cells and cultured for 24 hours. The effect of fibroblast growth factor (FGF) and transforming growth factor-beta(TGF-beta) on the MMC-treated cells was evaluated. In control group, osteoblasts were cultured in osteogenic media without exposure of MMC for 24 hours. Cell viability was measured using trypan blue staining method. RESULTS: As compared with control group, the growth of osteoblasts was inhibited by MMC (0.2 mg/ml) treatment, 30-minute treatment group demonstrated marked suppression twice as much as 5-minute treatment group. Growth rate of 0.2 mg/ml MMC-treated cells was highly suppressed to 7.7% of control and 0.02 mg/ml MMC-treated cells was inhibited to 15.4% in number. Growth rate of 0.002 mg/ml, 0.0002 mg/ml MMC-treated cells was diminished to 53.8%, 84.6% in number, respectively. Both growth factors had promotive effect on the growth of osteoblasts in 0.002 mg/ml MMC-treated cells, especially in TGF-beta. CONCLUSION: Osteoblasts which were treated for longer time and with higher concentration of MMC showed more severe suppression in growth rate. These results suggest that MMC could have some therapeutic effect on osteotomy site of dacryocystorhinostomy.
Cell Survival ; Dacryocystorhinostomy ; Fibroblast Growth Factors ; Intercellular Signaling Peptides and Proteins ; Mitomycin* ; Osteoblasts* ; Osteotomy ; Rabbits ; Transforming Growth Factor beta ; Trypan Blue

No abstract available.
Intercellular Signaling Peptides and Proteins*

No abstract available.
Intercellular Signaling Peptides and Proteins*

The aim of this study was to determine the optimal culturing media for human CD14+ monocytes and to evaluate whether these cells are capable of differentiating into vascular endothelial cells. Human monocytes isolated from peripheral blood were cultured for 1, 3, 7, 10 or 14 days in different media containing either 10% fetal bovine serum (FBS), 10% autologous donor serum (Auto), 10% FBS with interleukin-3 and macrophage colony stimulating factor (FBS-WF) or 10% Auto and the same growth factors (AU-WF). The cells were differentiated using endothelial cell conditioning medium (EC). Viability was measured using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and the cells were characterized by histology, immunohistochemistry and western blot analysis. Monocytes treated with Auto, FBS-WF or AU-WF medium generated a significant higher yield of vital cells after 7 days in culture compared with FBS-only medium (mean difference (MD)=0.318, P=0.01; MD=1.83, P=0.04; or MD=0.271, P=0.01 and MD=0.318, P=0.102). All tested media led to the differentiation of monocytes into macrophages, identified by CD68, especially in the FBS-WF medium (MD=+18.3% P=0.04). Differentiation into ECs caused a significant decrease in cell viability in all media. Endothelial cell markers, including CD31, CD144, VEGF, VEGF-R2 and CD34, could not be detected. Autologous serum significantly increases the yield of monocyte-derived cells with a higher effectiveness than commonly used FBS-only serum. There is no further benefit in culturing monocytes longer than 7 days. The cultivation of monocytes in the tested media leads preferentially to differentiation into macrophages. Differentiation into endothelial cells did not take place.
Blotting, Western ; Cell Survival ; Colony-Stimulating Factors ; Endothelial Cells ; Humans* ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; Interleukin-3 ; Macrophages ; Monocytes* ; Tissue Donors ; Vascular Endothelial Growth Factor A