AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanis...AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.METHODS: One hundred and eight Wistar rats were randomly divided into sham-operated control group (C)(n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nickend labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl 2 protein expression and distribution by immunohistochemical analysis.RESULTS: The rat survival rates in aFGF treated group were higher than group R (P＜0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17±3.49)%, (42.83±5.23)% and (53.33±6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67±6.95, 54.17±7.86, 64.33±6.47, respectively) (P＜0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in groupR during 2-12 h period after reperfusion.CONCLUSION: The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.展开更多
Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human larynge...Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human laryngeal cancer cellline Hep-2 and human vascular endothelial cells in vitro, cells got into the period of exponential phase of growth, was diviced into 3 groups：group I （control group）, group II （DDP group）, group III （Rg3 group）. Added to the Hep-2 cells Rg3 and DDP, made Rg3 final concentration was 300μg/mL, and DDP was 3μg/mL. 48 h later, specimens from sample to be done immunocytochemistry, and the protein of VEGF in Hep-2 cells to be detected. Col ecting Hep-2 cells supernatant, some was used to measure the protein level of VEGF in Hep-2 cells supernatant by ELISA. Some was used to culture HVEC. 24 h later, cellgrowth inhibition rate of human vascular endothelial was determined by MTT. Results：The protein level of VEGF was evi-dently higher in group I compared to group II and group III, it was not only in Hep-2 cells, but also in supernatant of Hep-2 cells. There was no significantly dif erent between group II and group III. MTT results showed that, the human vascular endothelial cellgrowth inhibition rate of group I was significantly lower than that of group II and group III （P〈0.05）. At the same time the HVEC growth inhibition rate of group II was significantly lower than that of group III （P〈0.05）. Conclusion：The inhibition to tumor angiogenesis of Rg3 is stronger than traditional chemotherapy drug cisplatin. It worke by reducing the biological ef ects of secreted VEGF, But the ef ecting worke by reducing the activity of secreted VEGF itself or af ecting endothelial function of VEGF receptor or some other ways to be further studied.展开更多
To investigate the proliferative ef fect of keratinocyte growth factor (KGF 2) on human adult keratinocytes. Methods: The standard medium was keratinocyte growth medium wit hout bovine pituitary extract (BPE), hydroco...To investigate the proliferative ef fect of keratinocyte growth factor (KGF 2) on human adult keratinocytes. Methods: The standard medium was keratinocyte growth medium wit hout bovine pituitary extract (BPE), hydrocortisone or epidermal growth factor ( EGF). Keratinocytes from a 48 year old subject were cultured and seeded on dis hes with standard medium of EGF in cell density of 2×10 4/32 mm 2. After 24 hours, the medium was replaced by the standard medium with 0, 4, 16, 125 and 50 0 ng/ml KGF 2, respectively. The standard medium with EGF was used as the posit ive control and the standard medium without EGF or KGF 2 was used as the negati ve controls. The growth of keratinocytes was monitored by 3 (4,5 dimethythiazo l 2 yl) 2,5 dipheyl tetrazolium bromide (MTT) assay and by photographs on day s 3, 5 and 7, respectively. Results: KGF 2 in concentrations of 4 500 ng/ml showed a sign ificant proliferative effect on days 5 and 7 as compared with that of the negati ve controls (P< 0.01 ). On day 3 the cells were prolifer ated to 1.5 2.5 fold, on day 5 to 3 5 fold and on day 7 to 3 12 fo ld in KGF 2 medium as that of the negative controls. The optimal response occur red when the concentration of KGF 2 was 125 ng/ml on day 7. Cell proliferation was also consistently higher in all KGF 2 concentrations as compared with that of the positive controls. Conclusions: KGF 2 has significant effects on the proliferatio n of adult keratinocytes, which are more effective than that of EGF. This study supports KGF 2 can improve the healing of chronic wounds in adults in clinic.展开更多
Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of...Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor （VEGF） expression of ECV304 cells, and VEGF, angiopoietin-1 （Ang-1）, platelet-derived growth factor （PDGF） and fibroblast growth factor 2 （FGF2） expression of WI-38 cells were assessed by 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-di-phenytetrazoliumromide （MTT） assay, viable cell counting, flow cytometry （FCM）, and enzyme-linked immunosorbant assay （ELISA）. Results： We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion： The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.展开更多
基金Supported by the National Basic Science and Development Programme, No. G1999054204the National Natural Science Foundation of China, No. 30170966, 30230370
文摘AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.METHODS: One hundred and eight Wistar rats were randomly divided into sham-operated control group (C)(n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nickend labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl 2 protein expression and distribution by immunohistochemical analysis.RESULTS: The rat survival rates in aFGF treated group were higher than group R (P＜0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17±3.49)%, (42.83±5.23)% and (53.33±6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67±6.95, 54.17±7.86, 64.33±6.47, respectively) (P＜0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in groupR during 2-12 h period after reperfusion.CONCLUSION: The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.
文摘Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human laryngeal cancer cellline Hep-2 and human vascular endothelial cells in vitro, cells got into the period of exponential phase of growth, was diviced into 3 groups：group I （control group）, group II （DDP group）, group III （Rg3 group）. Added to the Hep-2 cells Rg3 and DDP, made Rg3 final concentration was 300μg/mL, and DDP was 3μg/mL. 48 h later, specimens from sample to be done immunocytochemistry, and the protein of VEGF in Hep-2 cells to be detected. Col ecting Hep-2 cells supernatant, some was used to measure the protein level of VEGF in Hep-2 cells supernatant by ELISA. Some was used to culture HVEC. 24 h later, cellgrowth inhibition rate of human vascular endothelial was determined by MTT. Results：The protein level of VEGF was evi-dently higher in group I compared to group II and group III, it was not only in Hep-2 cells, but also in supernatant of Hep-2 cells. There was no significantly dif erent between group II and group III. MTT results showed that, the human vascular endothelial cellgrowth inhibition rate of group I was significantly lower than that of group II and group III （P〈0.05）. At the same time the HVEC growth inhibition rate of group II was significantly lower than that of group III （P〈0.05）. Conclusion：The inhibition to tumor angiogenesis of Rg3 is stronger than traditional chemotherapy drug cisplatin. It worke by reducing the biological ef ects of secreted VEGF, But the ef ecting worke by reducing the activity of secreted VEGF itself or af ecting endothelial function of VEGF receptor or some other ways to be further studied.
文摘To investigate the proliferative ef fect of keratinocyte growth factor (KGF 2) on human adult keratinocytes. Methods: The standard medium was keratinocyte growth medium wit hout bovine pituitary extract (BPE), hydrocortisone or epidermal growth factor ( EGF). Keratinocytes from a 48 year old subject were cultured and seeded on dis hes with standard medium of EGF in cell density of 2×10 4/32 mm 2. After 24 hours, the medium was replaced by the standard medium with 0, 4, 16, 125 and 50 0 ng/ml KGF 2, respectively. The standard medium with EGF was used as the posit ive control and the standard medium without EGF or KGF 2 was used as the negati ve controls. The growth of keratinocytes was monitored by 3 (4,5 dimethythiazo l 2 yl) 2,5 dipheyl tetrazolium bromide (MTT) assay and by photographs on day s 3, 5 and 7, respectively. Results: KGF 2 in concentrations of 4 500 ng/ml showed a sign ificant proliferative effect on days 5 and 7 as compared with that of the negati ve controls (P< 0.01 ). On day 3 the cells were prolifer ated to 1.5 2.5 fold, on day 5 to 3 5 fold and on day 7 to 3 12 fo ld in KGF 2 medium as that of the negative controls. The optimal response occur red when the concentration of KGF 2 was 125 ng/ml on day 7. Cell proliferation was also consistently higher in all KGF 2 concentrations as compared with that of the positive controls. Conclusions: KGF 2 has significant effects on the proliferatio n of adult keratinocytes, which are more effective than that of EGF. This study supports KGF 2 can improve the healing of chronic wounds in adults in clinic.
基金supported by the National Basic Research Program (973) of China (No.2005CB623906)the Medical Development Foundation of Soochow University (No.EE134702),China
文摘Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor （VEGF） expression of ECV304 cells, and VEGF, angiopoietin-1 （Ang-1）, platelet-derived growth factor （PDGF） and fibroblast growth factor 2 （FGF2） expression of WI-38 cells were assessed by 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-di-phenytetrazoliumromide （MTT） assay, viable cell counting, flow cytometry （FCM）, and enzyme-linked immunosorbant assay （ELISA）. Results： We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion： The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.
