ABSTRACT
Objective:To observe the impacts of herb-partitioned moxibustion,warm moxibustion and electroacupuncture on the basic fibroblast growth factor(bFGF)and collagen type Ⅰ(Col Ⅰ)in colons of rats with Crohn' disease(CD),and discuss the mechanism of acupuncture therapy on the intestinal fibrosis in CD.Methods:The model rats were developed by TNBS as multiple proinflammatory method.The rats were randomly divided into 5 groups:a normal group,a model group,a warm moxibustion group,an electroacupuncture group and a herb-partitioned moxibustion group.The treatments were carried out at Tianshu(ST 25)(bilateral)and Qihai(CV 6)in different treatments.The immunohistochemistry was used to detect the expression position of Col Ⅰ and bFGF.Results:The expressions of Col Ⅰ and bFGF in colons of rots in the model group significantly increased(compared with the normal group,P<0.01).After the herb-partitioned moxibustion,warm moxibustion and electroacupuncture,the expressions of Col Ⅰ and bFGF reduced markedly in the rats with CD(P<0.01).The expression of bFGF and Col Ⅰ in the colons had an obvious correlation in the Spearman rank correlation analysis.Conclusion:Acupuncture treatment reduced the abnormally high levels of expressions for Col Ⅰ and bFGF in colons.Col Ⅰ and bFGF participated in the fibrosis.Acupuncture treatment may reduce the bFGF expression in colons to regulate the excessive deposition,treating the intestinal fibrosis in CD.
ABSTRACT
Objective:To study the effect of a new medical biomaterial—bfgf-fibrin glue-amniotic membrane(bFAM) for corneal alkali burn and provide experimental foundation for the clinical application in the future. Methods: We use fibrin glue compounded with bFGF and amniotic membrane to prepare the bFAM. 45 New Zealand rabbits were used to make corneal alkali burn models and divided into 3 groups:bFAM group、amniotic membrane group、comparison group, each group have 15 rabbits. After operation, we observe the linical curative effect. 7d、14d and 28d after operation, 5 corneas were extracted for histopathological analysis. The expression of bFGF in corneas was measured by immunohistochemistry analysis. Results: After operation,bFAM group have better curative effect than amniotic membrane group in Inflammatory reaction、tissue repair and anti-CNV. The expression of bFGF in bFAM group is much higher than amniotic membrane group and comparison group in the day 7 afer operation, and in other time points the expression have not distinct difference in all groups. Conclusion: Compared with single layer amniotic membrane, bFAM shows better curative effect for corneal alkali burn.
ABSTRACT
· AIM: To explore the relationship between the expression of Fas/FasL and the apoptosis in retinal ischemia/reperfusion injury of rats, as well as the therapeutic effects of basic fibroblast growth factor (bFGF)on the ischemic retina.injury were made by transiently elevating introcular pressure. A total of 28 rats were divided into Normal Group and Operative Group. The latter were subdivided into 1, 6, 12, 24, 48 and 72h after reperfusion, in which the left eyes of the rats were in the ischemia/reperfusion groups and the right ones were in the treatment groups (bFGF intracameral injection). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and theexpression of Fas/FasL ligand was studied by strept avidin-biotin complex (SABC) immunohistochemistry.mai rats' retinae, but there were a significant number of TUNEL positive cells in 6-24h after transient ischemia followed by a decrease at 48h. The number of TUNEL positive cells reached a maximum at 24h after ischemia.The expression of Fas gradually increased as early as at 6h, reached a peak at 24h, then decreased at 48h. Similarly, the expression of Fas ligand was at peak in 24-48h in GCL and INL of retina. bFGF ministered before reperfusion inhibited apoptotsis and ameliorated the tissue damage. It also diminished Fas and FasL expression in ischemic/reperfused retina.siently elevated IOP induced apoptosis of cells in the retina. Fas/FasL may have an important role in the early events of the apoptotic pathways. bFGF can rescue RGCs from retinal ischemia/reperfusion injury through down-regulation of Fas and Fas ligand expression and may represent an important mechanism for therapeutic neuroprotection.
ABSTRACT
AIM To investigate if scallop (Placopecta magellanicus) skirt glycosaminoglycan (SS-GAG) inhibits the proliferation of vascular smooth muscle cell (VSMC) as heparin does so and to clarify its mechanism. METHODS The inhibitory effects of SS-GAG on the proliferation of rat thoracic aorta and abdominal aorta VSMC induced by fetal bovine serum (FBS) or basic fibroblast growth factor (bFGF) were determined by cell counting, crystal violet staining and MTT colorimetry. The effects of SS-GAG on the expression of proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor (PDGF) in VSMC proliferation induced by bFGF were evaluated by immunohistochemical technique (LSAB method) and computer image analysis system. RESULTSSS-GAG exerted antagonistic effects on VSMC proliferation induced by 20% FBS and 50 μg·L-1 bFGF at concentrations ranging from 50 mg·L-1 to 200 mg·L-1 and repressed the increasing expression of PCNA and PDGF. CONCLUSION SS-GAG significantly inhibits the proliferation of VSMC, which may be carried out through repression of PDGF and PCNA expression.
ABSTRACT
Objctive To explore the relationship between the expression of Fas/FasL and the apoptosis occurs in retinal ischemia/reperfusion injury of rats, as well as the therapeutic effects of bFGF on the ischemic retina. Methods The models of retinal ischemia/reperfusion injury was made by transient elevating introcular pressure. A total of 28 rats were divided into normal and operation group.The latter were subdivided into 1 hour, 6, 12, 24, 48 and 72 hours after reperfusion group, in which the left eyes of the rats were in the ischemia/reperfusion groups and the right ones were in the treatment groups (bFGF intracameral injection). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of Fas and Fas ligand was studied by strept avidin-biotin complex (SABC)immunohistochemistry. Results No positive cells were observed in the normal rats′ retinae, but there was a significant number of TUNEL positive cells in 6-24 hours after transient ischemia followed by a decrease at the 48th hour. The number of TUNEL positive cells reached a maximum at the 24th hour after ischemia. The expression of Fas gradually increased as early as when it was at the 6th hour, reached a peak at the 24th hour, and then decreased at the 48th hour. Similarly, the expression of Fas ligand was at peak in 24-48 hours in GCL and INL of retina. Conclusions Retinal ischemia-reperfusion after transient elevated IOP induced apoptosis of cells in the retina. Fas/FasL may play an important role in the early events of the apoptotic pathways. bFGF can rescue RGCs from retinal ischemia/reperfusion injury through downregulation of the expression of Fas/FasL and may represent an important mechanism for therapeutic neuroprotection.
ABSTRACT
Objective To investigate the differentiation from human mesenchymal stem cells (hMSC) into neuron-like cells. Methods hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with DMEM/BHA/DMSO or DMEM/monothioglycerol, respectively. Neuron-specific enolase (NSE), neurofilament (NF), nestin, glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. Results hMSC were expanded to be undifferentiated cells in culture for more than 10 passages. The isolated and cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. Simple method induced hMSC exhibiting a neuronal phenotype, with a positive expression of NSE, NF-M and nestin at 5 hours. But the neuron-like cells did not express the glial astrocyte marker GFAP. Conclusion It suggests that hMSC can be differentiated into neurons in vitro .
ABSTRACT
Objective To investigate the expression of the basic fibroblast growth factor (bFGF) in normal and degenerated human intervertebral discs in order to determine whether bFGF is related to degeneration of the intervertebral disc. Methods The specimens of the intervertebral discs from 11 male and 19 female patients undergone lumbar disc herniation surgery (observation group) and 6 patients with scoliosis following anterior release (control group) were harvested. The tissues were histologically observed to confirm their degenerated or normal status and then conducted for the expression of bFGF and its mRNA with immunohistochemistry and in situ hybridization. Results In the observation group, all the samples were found to be degenerated disc tissue, and the positive rate of the expression of bFGF was 90% (27/30) with immunohistochemistry and 20% (6/30) with in situ hybridization. In the control group, all the samples were shown to be normal disc tissues, and had negative expression of bFGF with immunohistochemistry and in situ hybridization. The positive rate of the expression of bFGF showed significant difference between the two groups. Conclusion The expression of bFGF showed significant difference between the degenerated and normal intervertebral discs, which indicated that the bFGF might promote proliferation of chondrocyte and synthesis of extracellular matrix in the degenerated discs as a proliferation stimulating factor.
ABSTRACT
To explore the mechanism of therapeutic effect of basic fibroblast growth factor (bFGF) in the treatment of blast-induced deafness, and to define its optimal clinical use, bFGF was infused into the guinea pig's cochlea, combined with intramuscular injection of bFGF after being exposed to explosion. The compound action potential (CAP) and auditory brainstem response (ABR) were measured in these animals. 125I labeled basic fibroblast growth factor ( 125I -bFGF) was injected intraperitoneal to the guinea pigs to observe whether it could pass through the blood-labyrinthine barrier. The results showed that bFGF infused to the cochlea might facilitate recovery of hearing loss following acoustic trauma. Basic fibroblast growth factor ( 125I -bFGF) intraperitoneally injected, could not pass through the blood-labyrinthine barrier. However intramuscular bFGF promoted the recovery of hearing, probably indirectly through the neuro-immunity network.
ABSTRACT
To investigate the effect of bFGF and sucralfate on the improvement of the quality of expanded skin flap, white piglets were employed to establish a continuous tissue expansion model. They were randomly divided into 3 groups: group 1: both bFGF and sucralfate were injected; group 2: both bFGF and normal saline was injected; and group 3(the control group):only normal saline was injected. Three days after completion of the expansion ,normal and expanded skin flaps were created at random to assess flap viability and stretch back .The results showed that the flap survival length in group 1 was significantly larger than that of the control group and normal random flap( P
ABSTRACT
Flap pedicled with the arteria epigastrica superficialis was transposed to primarily repair firearm wound after the rabbit's posterior limb was shotted. Recovery of the wound was observed and the content of bFGF mRNA in myoideum of the concussion area was measured by reverse transcroption polymerase chain reaction(RT PCR). All wounds repaired with transposition of fascia flaps attained primary healing and the content of bFGF mRNA in myoideum of the concussion area was higher than that in the control group. The result suggests that transposition of arterialized fascia flaps could primarily repair firearm wounds successfully and the high expression of bFGF is one of the causes.
ABSTRACT
To study the method and the optimal quantity of osteoblast transplantation to stimulate bone regeneration by transplanting 6～8 day old cultured osteoblasts and bFGF, together with osteoblast free calvaria of fetal rabbits into the defects of the rabbit radius. The fibroblasts were embedded in the type Ⅰcollagen in the concentrations of 10 7 ,10 6 ,10 5 , 10 4 /ml. Bone repair was evaluated by X ray, bone density,and scanning electron microscopy. Bone union ratio of the radius with transplantation of 10 7 ,10 6 ,10 5 , 10 4 /ml osteoblasts was 40%, 65%, 31 25%, 5 55% respectively at 4 weeks after transplantation, and 41 67%, 75%, 37 5%, 10% respectively at 8 weeks after transplantation. Bone density of radius defect with transplantation of 10 7 , 10 6 , 10 5 , 10 4 /ml osteoblasts was (0 112?0 018), (0 159?0 033), (0 122?0 039), (0 066?0 002) respectively at 6 weeks after transplantation and (0 150?0 059), (0 173?0 041), (0 145?0 023), (0 103?0 023) respectively at 8 weeks after transplantation, In 10 cases of bone non union, union was achieved by transplanting fetal osteoblast and basic fibroblast growth factor. This finding indicated that that transplantation of fetal osteoblasts could stimulate bone defect repair. The optimal concentration of osteoblasts implant was 10 6 cells /ml. It might be used in the treatment of bone non union.
ABSTRACT
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 measured 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
ABSTRACT
AIM: The role of protein kinase C(PKC) in the effect of Interleukin-6(IL-6) on basic fibroblast growth factor(bFGF) expression was investigated in rat vascular smooth muscle cells(VSMC). METHODS: Western-blotting was adopted to observe the variation of bFGF and its receptor type I isoforms expression. RESULTS: IL-6 increased all the three basic fibroblast growth factor isoforms in a dose-dependent (0-10.0 ?g/L) manner. The upregulatory activities peaked at 24 h as demonstrated . In addition, after exhaustion of intracellular phorbol ester-sensitive PKC, the upregulatory effects of IL-6 on bFGF exprssion in VSMC declined ( P
ABSTRACT
AIM: To observe whether metallothionein plays a role in cardiac protective effect of basic fibroblast growth factor on anoxic/reperfusion (A/R) injury in cultured cardiomyocytes and study the possible mechanism of cardiac protection by bFGF.METHODS: The present study made the model of myocyte A/R injury after having a 24 h incubation by bFGF(10 -10、10 -9、10 -8 mol/L) and bFGF(10 -9 mol/L)+PD 098059 respectively. We measured the levels of MT and MDA in myocytes, and the changes of LDH and protein in cultured medium. We also counted the number of viable cell in groups.RESULTS: The contents of myocardial MT were significantly increased after treatment by bFGF. The levels of MT in 10 -10 mol/L、10 -9 mol/L and 10 -8 mol/L bFGF treated groups increased 54%\, 62%\, 76% respectively, compared with the A/R group, and the number of viable cell were also greatly increased, LDH and protein leakage in cultured medium and MDA contents in myocyte were dramatically decreased in bFGF treated groups. All the protection were completely disappeared with the inhibition of MT production with PD 098059, the inhibitor of mitogen-activated protein kinase(MAPK).CONCLUSION: MT involves in the protection of bFGF in cultured cardiomyocytes. It might be related with activation of MAPKase.
ABSTRACT
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.
ABSTRACT
Objective To investigate the effects of GMl on inducing adult rat MSCs to the neural progenitor cells and their descendants. Methods Purified MSCs were induced by basic fibroblast growth factor (bFGF) alone, GMl alone or by combination of bFGF and GMl. After 3 days of incubation, fibronectin and collagen I were detected by immunocytochemistry, and nestin by immunofluorescence. Neuron-specific enolase ( NSE) , glial fibrillary acidic protein ( GFAP) and galactose cerebroside ( GalC) were detected by immunocytochemistry after 7 days of incubation. Results After induction by bFGF alone or combination of bFGF and GM1, some MSCs exhibited the phenotypes of neural progenitor cells in 3 days, and then exhibited neurons and astrocytes in 7 days. In these two groups, cells positive for fibronectin and collagen I were decreased markedly after 3 days of induction. At the same time, cells positive for nestin were increased markedly. After 7 days of induction, NSE and GFAP-positive cells were increased significantly (22. 28% ?2. 97% and 26. 65%?3. 17% ). Furthermore, the combination of bFGF and GMl showed a maximal increase (30. 35%?3. 51% and 32. 22%?2. 68% ). But the GMl alone had shown no inductive effects. Conclusion Combination of bFGF and GMl might synergistically improve the neural induction of MSCs, showing a promising route for the application of MSCs.
ABSTRACT
Objective To observe the reparative response and expression of basic fibroblast growth factor (bFGF) protein in the rotator cuff subjected to subacromial impingement. Methods Subacromial impingement of the infraspinatus tendon was experimentally created in 50 male SD rats by thickening the undersurface of the acromion with one platelike bony transplantation of the ipsilateral scapular spine. The contralateral shoulders that had undergone a sham operation were used as controls. The rats were sacrificed at 3, 7, 14, 28, and 56 th day, the whole shoulder joint was removed for detecting bFGF protein and the reparative response in the impinged infraspinatus tendon. Computer image analysis system were used to monitor the expression intensity and numbers of positive cells of bFGF protein. The OD scores and the size of area represent the expression intensity and numbers of positive cells respectively. Results All rats with experimental subacromial impingement showed an infraspinatus tear on the bursal side of the tendon. The shoulders in the control group were found intact without any alteration. There was proliferating cells in the fragmented tendons and vascularised connective tissue covering the area of ruptured area, whose source was the subacromial bursa. Few tenocytes and bursal cells expressed bFGF protein in unwounded tendons. In contrast, tendons subjected to impingement exhibited an increased signal for bFGF protein in both resident tenocytes concentrated along the epitenon and infiltrating fibroblasts and inflammatory cells from the subacromial bursa. Conclusion The bFGF protein is upregulated during tendon healing and the subacromial bursa is the main source of both bFGF secreting and rotator cuff repair; one should preserve as much as possible the subacromial bursa.
ABSTRACT
Objective To study the biological functions of growth factors in the process of osteoblasts allografting and its acceleration in fracture healing in osteoporosis rats. Methods The fracture model of rat osteoporosis was established and then cell grafting was performed. Immunnohistochemistry and in situ hybridization were utilized to examine the expression of bFGF and bFGF mRNA during the process of fracture healing, the image was also analyzed. Results In the experimental group, bFGF positive cells were seen after 7 days after cell grafting, and peaked in chondrocytes about 14 day (384 65?8 60) after grafting. bFGF began declining after 21 days (286 24?2 30) and disappeared after 56 days. However in the control group, obvious high quanty was not seen and being only 125 33?4 50 even peaked at 21st day. Conclusions bFGF not only plays a very important role in bone formation but also enhances circulation formation in the fracture areas. It also accelerates the fracture healing in osteoporosis rats.
ABSTRACT
Objective To further confirm the Efficacy and safety of tadenan (Pygeum africunum extract) in the treatment of benign prostatic hyperplasia (BPH) in larger population in China. Methods In this open multicenter study,a total of 3 508 patients with symptomatic BPH received oral tadenan of 50 mg twice daily for 8 weeks. Results IPSS,maximum urinary flow and post voiding volume were 19.57? 14.93 ,(13.52?5.36)ml/s and (44.58?30.95)ml four weeks after the treatment of tadenan and were 13.31?11.86,(15.58?5.98)ml/s and (32.64?24.23)ml eight weeks after the treatment,both improved significantly comparing to those before the treatment ( P
ABSTRACT
AIM: To observe whether metallothionein plays a role in cardiac protective effect of basic fibroblast growth factor on anoxic/reperfusion (A/R) injury in cultured cardiomyocytes and study the possible mechanism of cardiac protection by bFGF.METHODS: The present study made the model of myocyte A/R injury after having a 24 h incubation by bFGF( 10-10、10-9、10-s mol/L) and bFGF( 10-9 mol/L) + PD098059 respectively. We measured the levels of MT and MDA in myocytes, and the changes of LDH and protein in cultured medium. We also counted the number of viable cell in groups. RESULTS: The contents of myocardial MT were significantly increased after treatment by bFGF. The levels of MT in I0-l0 mol/L、10-9 mol/L and 10-8 mol/L bFGF treated groups increased 54 %、 62%、 76% respectively, compared with the A/R group, and the number of viable cell were also greatly increased, LDH and protein leakage in cultured medium and MDA contents in myocyte were dramatically decreased in bFGF treated groups. All the protection were completely disappeared with the inhibition of MT production with PD098059, theinhibitor of mitogen- activated protein kinase(MAPK). CONCLUSION: MT involves in the protection of bFGF in cultured cardiomyocytes. It might be related with activation of MAPKase.