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[Objective]To investigate the effect of transforming growth factor ?1(TGF-?1) genes transfection on the proliferation and differentiation of chondrocyte by adenovirus vector,and to observe the quality in repair cartilage defect through constructing tissue-engineered cartilage.[Method]Using replicat ion defective adenovirus Adeno-X~(TM) as a carrier,Articular chondrocytes were transferred by high titers level recombinant adenovirus taking TGF-?1 genes.Ultromicrostructure,prolife ration anddifferentiation were observed by light and electron microscope,flow cytometry and so on.Induce expression of exogenous genes coding protein and cartilage cell phenotype were detected by Immunocytochemistry and Northern blot.Gross observation,histology and effect classification were used to evaluate results in repair cartilage defect by combi nation of infected chondrocytes and bone matrix gelatin(BMG).[Result]Immunocytochemistric staining showedthe expression of exogenous gene coding proteins after infection,and procollagen Ⅱ mRNA in cells was detected increasingly by Northern blot.TGF-?1 stimulated proliferation of chondrocytes in primary cultured,and resulted in recruitment of articular chondrocytes into S-G2/M phase.Chondrocytes seeded onto BMG showed high level of proliferation.Transplantation in vivo showed: cartilage defect was repaired satisfactorily by Combination of BMG and infected chondrocytes,and the structure of repair tissue got close to normal articular cartilage.[Conclusion] Articular chondrocytes prolifera tion could be stimulated by adenovirus vector taking TGF-?1 genes,which could be used to constructed tissue-engineered cartilage,and applied in repairing joint cartilage defect.
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Objective To study the mechanism of transforming growth factor ?(TGF-?) and its receptor in the pathogenesis of hepatocellular carcinoma and to explore its clinical significance.Methods 36 hepatocellular carcinoma tisssues and 36 corresponding cancer-adjacent normal tissues were investigated by immunohistochemistry for the expression of TGF-? and its receptor.Results The expression of TGF-? was distributed mainly in cytoplasm,while its receptor mainly in cellular membrane and cytoplasm.The expression of TGF-? in hepatocellular carcinoma was higher than that of normal tissues(P
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[Objective] To investigate the effect of Biejia Jian Decoction on serum transforming growth factor ?1 (TGF-pl) , tumor necrosis factor a (TNF-?), lipid peroxidation ( LPO) and superoxide dismutase ( SOD) in patients with hepatic fibrosis (HF) induced by chronic hepatitis B. Its effect was compared with lamivudine. [Methods] Fifty-three cases of HF were randomized to two groups: group A (n = 27) was treated with Biejia Jian Decoction and group B ( n -26) with lamivudine for oral use. Twenty volunteers served as the normal controls (group C) . Serum levels of TGF-?1, TNF-?, LPO and SOD were observed before and after treatment. [Results] After treatment, serum levels of TGF-?1 and TNF-a were decreased markedly in group A (P 0.05) . Serum LPO level was lower and SOD level higher in groups A and B than those in group C before treatment ( P 0.05) but the level in group B differed from that of groups A and C (P 0.05) but the level in group B did not decrease and differed from that of groups A and C ( P
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Objective To introduce transforming growth factor ?(TGF ?) and the relationship between TGF ? and graft rejection. Methods Relevent articles in recent years were reviewed.Results The immunodepressive function of TGF ? could resist transplant organ rejection injury in early postoperative period ; meanwhile TGF ? also caused fibroblast migration and promoted matrix deposition by increasing collagen production and decreasing collagen breakdown via inhibition of collagenases,which resulted in transplant organ fibrosis and arteriosclerosis, gene polymorphisms of the TGF ? were associated with it. Moreover,ischemia reperfusion injury and immunodepressive drug also affected production of TGF ?.Conclusion TGF ? as a pleiotropic and multifunctional cytokine contributes to the development of acute and chronic rejection.
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Objective To investigate the role of interferon ?on smooth muscle cells proliferation and migration after balloon injury. Methods Animal model of rabbit iliac artery balloon injury was set up, smooth muscle cells derived from injured artery were cultured. Smooth muscle cells were divided into four groups ( control, TGF ? 1, IFN ?, TGF ? 1 and IFN ?).Cells from each group were treated with medium or TGF ? 1 (10 ng/ml) and/or IFN ?(500 u/ml) for 72 h separately. Smooth muscle cell proliferation was determined by cell count and MTT, migration of smooth muscle cells was also detected. Matrix metalloproteinase 2 was also detected by zymography. Results Our results showed that, compared with group control(2.875?0.323?10 5 cells/ml,279.9?8.129 ?m), TGF ? 1 increased cell count (4.188?0.239?10 5 cells/ml, P
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Objective To analyze the actions of transforming growth factor ? 1 (TGF-? 1) and tumor necrosis factor ?(TNF-?) in the liver fibrosis formation in chronic viral hepatitis patients and liver cirrhosis patients.Methods Serum levels of TGF-? 1 and TNF-? were measured by ELISA in 107 patients with chronic liver diseases,liver biopsy was performed in 23 patients in order to identify pathological stages under optic microscope.Results The serum levels of TGF-? 1(?g/L) in mild,middle and severe groups of chronic viral hepatitis and liver cirrhosis group were 14 2?5 9,20 1?7 0,30 2?6 7 and 32 6?7 5 respectively.The serum levels of TNF-? (pg/L) were 6 1?3 2,29 8?18 6,57 3?22 5 and 96 7?38 2 respectively.The serum levels of above two markers increased with hepatitis progression.The results of liver biopsy revealed that the changes in these two markers were related to liver cirrhosis degree.Conclusions TGF-? 1 and TNF-? are related to liver cirrhosis formation.
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Objective To evaluate the expression of TGF? 1 in rat testes after vas deferens ligature and release again. Methods Ninety rats were randomized into 3 groups:vas deferens ligature in 35 rats,release again after ligature in 20,sham-operation in 35.Immunohistochemical staining and hybridization in situ were used to detect the expression of TGF? 1. Results The expression of TGF? 1 in vas deferens ligature group was higher than that in sham-operation group after 8 weeks ( P
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Objective To investigate the influence of the aberrant protein expression and mRNA transcription of transforming growth factor ? 1(TGF? 1) on the biological behavior of human bladder transitional cell carcinomas. Methods The expression of TGF? 1 was investigated in 74 specimens of TCCs by SP immunohistochemistry staining, and the level of TGF? 1 mRNA were determined in 43 cases of TCCs by the method of quantitative RT PCR. Results The positives expression rate of TGF? 1 was 89.2%. Superficial tumors had lower overexpressive rate of TGF? 1(33.3%) than the invasive TCCs(83.8%), P
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Objective To investigate the protective effect of Quercetin on kidneys in diabetic rats. Methods STZ induced diabetic rats were given quercetin 100 mg?kg -1 ?d -1 for 8 weeks. Urinary albumin excretion rate (UAER) was measured by radioimmunoassay. The changes of creatinine clearance rate (Ccr) and glomerular protein kinase C (PKC) activities were determined. The expression of TGF ? 1 mRNA of renal cortex in diabetic rats were determined by RT PCR analysis. The glomerular changes were also observed morphologically. Results In untreated diabetic rats, Ccr, UAER, kidney weight/body weight and PKC activity in renal glomeruli were significantly increased, the expression of TGF ? 1 mRNA in renal cortex was elevated, and glomerular hypertrophy existed. After Quercetin treatment, Ccr, UAER, PKC activity and the expression of TGF ? 1 mRNA were markedly reduced as compared with those of untreated diabetic rats in 2 and 8 weeks, no significantly abnormal changes in kidney morphology were observed in Quercetin treated group. Conclusion Quercetin ameliorates early diabetic renal hyperdynamic abnormality via inhibiting PKC activity, in which inhibiting of TGF ? 1 production seems to be involved. Reduction of the PKC activity is important in preventing or delaying the development of diabetic nephropathy.
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Objective To study the effects of transforming growth factor ?1 (TGF ?1) and transforming growth factor ? (TGF ?) on the growth and invasive potentials in human bladder tumor cells. Methods The effects of TGF ?1 and TGF ? on the growth of EJ cell line and the effects on MMPs and TIMP 2 were studied by means of MTT,Western Blot and RT PCR methods. Results (1)TGF ?1 and TGF ? tended to inhibit the growth of EJ cells but statistically nonsigficant.(2)Higher levles of MMP 9 mRNA but lower levels of MMP 2,TIMP 2,MT1 MMP mRNA were found in EJ cells following the treatment with TGF ?1.The same was true for the expression of MMP 2,TIMP 2 mRNA in TGF ? groups.However,MMP 9 mRNA was not found in both TGF ? groups and the control groups. (3)TGF ?1 ( 0.1 , 1.0 ng/ml) enhanced MMP 2 protein but not the TIMP 2 protein,while TGF ?1 ( 5.0 , 10.0 ng/ml)decreased TIMP 2 protein but not on MMP 2.In TGF ? groups,when the concentration was 1.0 , 5.0 , 10.0 ng/ml,TIMP 2 protein expression was decreased but MMP 2 did not.When the concentration reached 100.0 ng/ml,it increased MMP 2 protein level,not the TIMP 2 protein. Conclusions TGF ?1 and TGF? do not inhibit the proliferation of EJ cells whereas the enhanced invasiveness and metastasis may be associated with regulating the expression of MMPs and TIMP 2.
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Objective To investigate the influence of transforming growth factor ? 1 to expression of tumor necross factor ?(TNF ?) after cerebral ischemic reperfusion injury.Methods Using thread embolism method to develop the model of focal cerebral ischemic reperfusive injury in rats, different dose of TGF ? 1 or 0 9% NaCl was injected intracerebroventricularly.Neurological functional dificit scales in the different dose of TGF ? 1 groups and the expression of TNF ? were observed Results In both big and small dose of TGF ? 1 groups,the expression of TNF ? decreased after cerebral ischemic reperfusion,and the neurological functional dificit scales were significantly lower than those of the control.There was no significant difference between the group of big and small dose of TGF ? 1 Conclusion TGF ? 1 may have a protective effect on cerebral ischemic reperfusive injury by inhibiting expression of TNF ?,and it may be irrelative with the dose
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Objective To understand the expressions of transforming growth factor ?1(TGF-?1) and its receptors(T?R) in villi and decidua during early pregnancy and their effects on early embryo development, and explore the mechanism of spontaneous abortion. Methods By immunohistochemical technique, expressions of TGF-?1 and its receptors were determined in villi and decidua from 10 cases of spontaneous abortion women after in vitro fertilization and embryo transfer (IVF-ET group), 20 cases of spontaneous abortion women (spontaneous abortion group), and 20 cases of normal early pregnancy women (control group). Results TGF-?1 and its receptors had high expressions in decidua cells, villus glands and interstitial cytoplasm in all three groups. The average light density of the expressions of TGF-?1 in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.167 and 0.199, 0.198 and 0.201, 0.277 and 0.274, respectively. The intensity of T?R-Ⅰ in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.144 and 0.150, 0.202 and 0.201, 0.238 and 0.281, respectively. And the expressions of T?R-Ⅱ in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.199 and 0.145, 0.153 and 0.156, 0.300 and 0.301, respectively. The differences between the control group and both abortion groups were all significant (P
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Objective To determine the effects and mechanisms of somatostatin analogues (sandostatin) on pancreatic repair and regeneration in caerulein induced pancreatitis. Methods Acute pancreatitis was induced by intra abdominal infusion of caerulein in rats, sandostatin was administered intra abdominally at the time of induction of pancreatitis and 24, 48 and 72 hours after. Rats were sacrificed at 6, 24, 48, 72 and 96 hours after the operation. The mRNA expression for Transforming growth factor ? 1 (TGF ? 1) was evaluated by reverse transcription polymerase chain reaction, pancreatic tissue DNA synthesis was measured by 3H thymidine method in vitro and protein content was detected by Lowry's method. Results The serum amylase level was decreased significantly in the sandostatin treated group. Expression of TGF ? 1 mRNA was undetectable in the normal pancreas and the treated group at 6 hours. TGF ? 2 was observed at 24 hours after the induction of pancreatitis, reaching maximum at 72 hours. It could be detected in the sandostatin treated group at 6 hours, reaching maximum at 24 hours, the expression of TGF ? 1 was increased significantly in the sandostatin treated group as compared with the non treated group at 24, 48 hours. Pancreatic tissue DNA synthesis showed a significant decrease during the first 72 hours following the induction of pancreatitis and a marked increase was observed at 96 hours after treatment with sandostatin. Within 48 hours of the induction of pancreatitis, total protein content in pancreatic tissue declined, and there was a significant increase in the sandostatin treated group at 48 hours, reaching maximum at 96 hours. Conclusions Effects of somatostatin analogues (sandostatin) on pancreatic tissue regeneration in acute pancreatitis in rats might be attributed to the enhancement of TGF ? 1 gene expression which subsequently stimulates formation of extracellular matrix components, increases protein content and DNA synthesis, thus accelerates pancreatic repair and regeneration.
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Objective To investigate the influence of endogenous transforming growth factor ? 1 (TGF? 1) on the cell cycle regulation and proliferation of bladder cancer. Methods A constructed replication defective retroviral vector pRevT? AS,which carried antisense RNA of TGF? 1,was transfected to a bladder cancer cell line EJ.The proliferation and clone formation of transferred cells were observed in vitro,and the alteration of cell cycle was also detected by flow cytometric analysis. Results TGF? 1 antisense RNA was transferred into EJ cell and expressed efficiently.After the inhibition of target gene expression in EJ cells,the reduced growth and clone formation rates were demonstrated,and the proliferative indexes were decreased by 12%.The ratios of G 0 and G 1 stage cells to the antisense RNA transfected EJ cells were increased,simultaneously,the ratio of S stage cells to the antisense RNA transfected EJ cells ratios were decreased,compared with the control group. Conclusions The proliferative mechanism of endogenous TGF? 1 in bladder cancer cells is to stimulate the G 1 to S stage transition.
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Objective To explore the effects of simvastatin and cilazapril on the expression of transforming growth factor-?_1 (TGF-?_1) and insulin-like growth factor-Ⅰ (IGF-Ⅰ) in the cultured human glomerular mesangial cells. Methods Human embryo glomerular mesangial cells were cultured in media with lower (5.6 mmol/L) or higher (30 mmol/L) glucose concentrations. Forty-eighthoursafteraddingsimvastatin(10 ?mol/L)and/or cilazapril (10 ?mol/L) to the cultured media, the concentrations of TGF-?_1, fibronection, laminin, type Ⅳ collagen proteins in the supernatant of cultured mesangial cells were determined by ELISA and radioimmunoassay and the expressions of TGF-?_1 and IGF-Ⅰ mRNA in cultured mesangial cells were also evaluated by RT-PCR. Results Compared with lower glucose control, the mesangial cells in the medium with higher glucose concentration showed excessive proliferation and higher expressions of TGF-?_1 and IGF-Ⅰ mRNA, and the levels of TGF-?_1, fibronection, laminin, and type Ⅳ collagen in the supernatant were also significantly increased. The expressions of TGF-?_1 and IGF-Ⅰ mRNA in the mesangial cells and the concentrations of TGF-?_1 and the extracellular matrix (ECM) proteins in the supernatant were all decreased after addition of simvastatin and cilazapril. Combination of simvastatin and cilazapril resulted in more profound suppressive effect on the expression of TGF-?_1 mRNA than either of them alone. Conclusion High concentration of glucose stimulates the cultured human mesangial cells excessively to express the TGF-?_1, IGF-Ⅰ and ECM proteins, and the high glucose-induced changes are suppressed by either simvastatin, cilazapril alone or combined treatment.
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Objective To study the regulatory effect of TGF ? 1 on the expression of multiple genes in prostatic stromal cells in vitro. Methods The primary culture of prostatic stromal cells (including fibroblasts and smooth muscle cells ) have been established and cultured to 4～6 passages. Then the cells were cultured in the medium with various concentration of TGF ? 1(0.01, 0.10, 1.00 and 10.0 ng/ml)for 48h. By semi quantitative RT PCR method, the androgen receptor(AR), TGF ? 1, bFGF and smoothlin mRNA were measured. Results The expression of AR could be stimulated by low concentration of TGF ? 1 ( P
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Objective:To investigate the effect of TGF-? on the expression of type Ⅰ inositol 1,4,5-triphosphate receptor in WB rat liver epithelial cells.Methods:The expression of type Ⅰ inositol 1,4,5-triphosphate receptor protein and mRNA were detected by Western blot and RT-PCR in WB rat liver epithelial cells in vitro stimulating with TGF-?.Results:The expression level of type Ⅰ inositol 1,4,5-triphosphate receptor protein and mRNA in WB rat liver epithelial cells were both increased after stimulating with TGF-? in several time points and reached the highest at 8 h stimulated, and then decreased.Conclusion:TGF-? enhance the IP_3R1 protein and mRNA expression in WB rat liver epithelial cells.
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Objective: To investigate the relationship between the Smad4-independent pathway of TGF-?1 and drug-resistance of pancreatic cancer. Methods: The sensitivities of Smad4 homozygous-deleted pancreatic cancer cell line BxPC3 to different kinds of anti-cancer drugs (5-Fu, Gemzar, Oxaliplatin, Cisplatin, CPT-11 and Epirubicin) were observed by MTT assay before and after they were transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 (5 and 10 ng/ml) solution. Western blot was used to detect p170 protein expression after stimulation with different concentrations of TGF-?1. Results: Cisplatin had the most powerful killing effect on BxPC3 cells, followed by Oxaliplatin, 5-Fu and CPT-11 with moderate effect and Gemzar and Epirubicin with the least effect. Cells transfected with full-length cDNA of TGF-?1 or treated with TGF-?1 solution became less sensitive to Cisplatin. Western blot revealed upregulation of p170 expression by TGF-?1. Conclusion: The Smad4-independent pathway of TGF-?1 can increase the drug resistance of pancreatic cancer cells through upregulating expression of p170.
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Objective To block the bio function of TGF ? by the application of anti TGF ? polyclonal antibody, and observe its efficacy of prevention of abdominal adhesion. Methods The postoperative abdominal adhesion model was established in SD rats. The drugs were administrated by abdominal injection with saline (control group), sodium hyaluronate( HA group), and varied dosage of anti TGF? (anti TGF? groups) respectively. The adhesion was scored 21days later, while 30 of them were executed on the day 3 and day 10 after operation respectively. The expression of TGF ? was checked by immunohistochemistry in the samples obtained from the adhesion sites. Results The score of adhesion in anti TGF ? group (2.4?0.99) was significantly lower than that in control group (6.0?1.25) and HA group (3.4?1.03); in different dosage of anti TGF, the 50?g group showed its economical efficacy; in the control and HA groups the expression of TGF ? had a time dependent manner, which reachs to maximum in the day 3, and could be reduced by antibody. Conclusions The polyclonal antibody of TGF ? shows the power to prevent the postoperative abdominal adhesion in animal model, the mechanism of which is due to inhibition of TGF ? expression.
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Objective To investigate the influence of vitamin E on type Ⅰ collagen, messenger RNA expression of transforming growth factor ? 1 (TGF? 1) and tissue inhibitor of matrix metalloproteinases-2 (TIMP 2) in hepatic stellate cells (HSC). Methods HSC s were cultured in vitro, the effect of vitamin E on type Ⅰ collagen was observed by mean of immunohistochemistry; mRNA expression of TGF? 1 and TIMP 2 were investigated with in situ hybridization. Results In experiment group, vitamin E markedly inhibited expression of type Ⅰ collagen of HSC as compared with control group, but could not suppress mRNA expression of TGF? 1 and TIMP 2.Conclusions Anti-fibrotic effect of Vitamin E may be implemented through inhibiting the expression of type Ⅰ collagen.