mRNA Expression Levels Of TGF-β1 Research Articles

E0040 The expression of signal transduction pathway of TGF1Smad in the rat cardiac tissue with type 2 diabetes and intervention by atorvastatin

ObjectiveObservation on changes of signal transduction pathway of TGF-β1/Smad in the course of myocardial fibrosis in the rat with type 2 diabetes and preventive effect of atorvastatin.MethodsThe experimental type 2...

Rheumatic valvular surgery and concomitant radiofrequency Maze procedure: TGF-β1 mRNA expression predicting the efficacy

To investigate the relationship of the transforming growth factor beta1 (TGF-β1) mRNA expression in atrial myocardium and the effectiveness of radiofrequency Maze procedure in patients with rheumatic valvular disease (RHD) and permanent atrial fibrillation (AF). Between January 2008 and September 2008, 40 patients with RHD and AF underwent a radiofrequency Maze procedure with concomitant valvular surgery. The patients were assigned to normal sinus rhythm (SR) group (group A) and persistence AF group (group B) according to the results of the 6-month follow-up. Another 10 patients with SR and RHD undergone valvular surgery alone were assigned to control group (group C). Left atrial appendage were obtained in all patients. Expressions of TGF-β1 mRNA were detected by semiquantitative RT-PCR technique. CVF-I and CVF-III were observed by sirius red staining. At 6-month follow-up, there were 28 patients in group A and 12 in group B. Patients in group A and group B had higher mRNA expressions of TGF-β1, CVF-I and CVF-I/CVF-III compared with group C (P < 0.05). Also, the group B had higher mRNA expressions of TGF-β1, CVF-I and CVF-I/CVF-III than group A (P < 0.05). The patients who had return of functional atrial contraction in group A had lower mRNA expression than the non-return patients (39 ± 12 vs. 60 ± 12, P < 0.05). The TGF-β1 mRNA expression had a correlation with both the contents of CVF-I and left atrial diameter (r = 0.786, P < 0.05; r = 0.858, P < 0.05). Multiple logistic regression analysis revealed that the risk factors which independently associated with the postoperative persistence of atrial fibrillation at 6-month follow-up includes mRNA expression levels of TGF-β1 (OR = 1.13, 95%CI 1.05 - 1.18, P = 0.031), CVF-I (OR = 1.07, 95%CI 1.00 - 1.13, P = 0.037) and lett atrial diameter (OR = 2.23, 95%CI 1.08 - 4.59, P = 0.042). The atrial TGF-β1 mRNA expression level could predict the persistence of AF and the return of the functional atrial contraction at 6-month follow-up in patients who underwent rheumatic valvular surgery and concomitant radiofrequency Maze procedure.

−509C&gt;T polymorphism in the TGF-β1 gene promoter is not associated with susceptibility to and progression of colorectal cancer in Chinese

Colorectal cancer is common, accounting for nearly 10% of all cancers. Transforming growth factor-β1 (TGF-β1) is a pleiotropic cytokine that has been implicated in the pathogenesis of colorectal neoplasia. The most studied -509C>T polymorphism of TGF-β1 gene has been associated with various kinds of cancer. This study investigated the association between this genetic variant and the risk and/or progression of colorectal cancer. A case-control study was carried out of 150 colorectal cancer cases and 503 healthy controls. DNA was extracted from blood cell nuclear materials, and -509C>T polymorphism in the TGF-β1 gene promoter was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Colorectal cancer tissues (n = 70) were obtained from the studied cases for measurement of TGF-β1 mRNA expression levels. We also assessed the plasma TGF-β1 levels of cases (n = 88) and healthy subjects (n = 120). The TGF-β1 producer genotype, -509TT, was not associated with an increased risk of colorectal cancer compared with other genotypes. Colorectal cancer patients especially those with a more aggressive disease behaviour were more frequently associated with C allele. The results suggest that TGF-β1 -509C>T polymorphism is not associated with either an increased risk or progression of colorectal cancer.

Change of Peripheral Blood Mononuclear Cells IFN-γ, IL-10, and TGF-β1 mRNA Expression Levels With Active Human Cytomegalovirus Infection in Orthotopic Liver Transplantation

Aim To analyze the expression levels of interferon gamma (IFN-γ), interleukin 10 (IL-10), and transforming growth factor beta 1 (TGF-β1) mRNA in peripheral blood mononuclear cells (PBMCs) in liver transplanted recipients with active HCMV infection. Methods PBMCs were isolated from 20 liver transplanted recipients with active HCMV infection and 20 recipients without HCMV infection. The expression levels of IFN-γ, IL-10 and TGF-β1 mRNA in PBMCs were measured by TaqMan real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The results were compared with that from 20 healthy individuals. Results The expression level of TGF-β1 mRNA was significantly increased in the active HCMV infection group compared with that in stable group or healthy group ( P < .001). The expression level of IL-10 mRNA was significantly increased in the active HCMV infection group compared with the healthy group ( P = .001). However, the IFN-γ mRNA expression level was significantly decreased in the active HCMV infection group compared with that in the stable group or the healthy group ( P < .05). Conclusion Cytokine production plays a role in the HCMV infection. This may provide an important clue to a better understanding of the pathogenesis in liver transplanted recipients with active HCMV infection.

Immune self-tuning in renal cell carcinoma patients

e22069 Background: Renal cell carcinoma (RCC) cells can inhibit protective antitumor immunity by secretion of immunosuppressive factors leading to the induction of regulatory T cells. The objective of this study was to investigate the prognostic impact of mRNA expression levels of IL10, TGFβ and forkhead box transcription factor (FoxP3) mRNA in peripheral blood mononuclear cells (PBMCs) of metastatic RCC patients before receiving treatment with sorafenib. Methods: PBMCs of 46 patients were assessed for their expression levels of TGFβ, IL10 and FoxP3 by quantitative RT-PCR. Clinical features considered included ECOG performance status, hemoglobin, alkaline phosphatase, and calcium concentrations. Disease evaluation was performed every 8 weeks following RECIST criteria. Relationship between pre-treatment factors and survival were examined in univariate analyses and subsequently by multivariate analysis using a stepwise Cox regression model. Results: In contrast to FoxP3, mRNA expression levels of IL10 and TGFβ were significantly higher in the 46 RCC patients compared to healthy volunteers: Median expression levels [ratio marker /housekeeping gene PBGD] were 5.56E-05 vs 2.05E-04 (P=0.034) for IL10 and 7.38E-02 vs 3.04E- 01 (P=0.023) for TGFβ. Univariate analysis revealed a negative prognostic influence of IL10 on progression free survival (p=0.04) and on overall survival, although not significant (P= 0.063). Surprisingly, high TGFβ and FoxP3 expression levels had a positive influence on progression free (P&lt;0.001 and P=0.047, respectively) and overall survival (P&lt;0.001 and P= 0.031, respectively). In the multivariate analysis low ECOG performance status and high TGFβ mRNA expression levels were independently associated with worse progression free (P=0.001 and P=0.054,) and worse overall survival (P=0. 006 and P&lt; 0.001, respectively). Conclusions: RCC caused an immune-suppressive phenotype in PBMC characterized by increased mRNA expression levels of IL10 and TGFβ. Surprisingly, in contrast to IL10, a high TGFβ mRNA expression level was an independent good prognostic factor. Whether this observation can be attributed to recently described immune promoting functions of TGFβ needs to be determined. No significant financial relationships to disclose.

Natriuretic peptide/natriuretic peptide receptor-A (NPR-A) system has inhibitory effects in renal fibrosis in mice

Object This study was designed to examine whether natriuretic peptide/natriuretic peptide receptor-A (NPR-A) system attenuates renal fibrosis in a unilateral ureteral obstruction (UUO) model and also examined the mechanism involved. Methods Three groups were studied: untreated UUO in wild-type mice; untreated UUO in NPR-A KO mice; and ANP treated (0.05 µg/kg/min) UUO in wild-type mice. We measured histological and immunohistochemical findings (α-SMA and F4/80), tissue cGMP levels, various mRNA expression levels by real-time PCR analysis, and transcription factor levels (AP-1 and NF-kappaB) in renal tissue. Results Compared with wild-type UUO mice, NPRA-KO UUO mice had abnormal morphological findings (fibrous area: + 26%, α-SMA expression: + 30%) with lower tissue cGMP levels and increases in the mRNA expression levels of TGF-β, collagen I, collagen III, PAI-1, renin and angiotensinogen, whereas there were no differences in F4/80 positive cells or the mRNA expression levels of ICAM-1, osteopontin, or MCP-1 between the two groups. In contrast, ANP pre-treatment significantly improved morphological changes with increase of tissue cGMP levels and reduction in the mRNA expression level of TGF-β, collagen I, collagen III, PAI-1, ICAM-1, osteopontin, MCP-1, renin, and angiotensinogen. NPRA-KO UUO mice had higher AP-1 levels than wild-type UUO mice and ANP pre-treatment reduced AP-1 and NF-kappaB activity. Conclusion The endogenous natriuretic peptide/NPR-A system may inhibit renal fibrosis partly via inhibition of the angiotensin/AP-1/TGF-β/collagen pathway and exogenous ANP pre-treatment may inhibit it partly via both the angiotensin/AP-1/TGF-β/collagen and NF-kappaB/inflammatory pathways.

Black seed oil ameliorates allergic airway inflammation by inhibiting T-cell proliferation in rats

The black seeds, from the Ranunculaceae family, have been traditionally used by various cultures as a natural remedy for several ailments. In this study, we examined the effect of black seed oil as an immunomodulator in a rat model of allergic airway inflammation. Rats sensitized to ovalbumin and challenged intranasally with ovalbumin to induce an allergic inflammatory response were compared to ovalbumin-sensitized, intranasally ovalbumin-exposed rats pretreated with intraperitoneally administered black seed oil and to control rats. The levels of IgE, IgG1 and ova-specific T-cell proliferation in spleen were measured by ELISA. The pro-inflammatory cytokine IL-4, IL-5, IL-6 and TGF-β1 mRNA expression levels were measured by reverse transcription polymerase chain reaction. The intraperitoneal administration of black seed oil inhibited the Th2 type immune response in rats by preventing inflammatory cell infiltration and pathological lesions in the lungs. It significantly decreased the nitric oxide production in BALF, total serum IgE, IgG1 and OVA-specific IgG1 along with IL-4, IL-5, IL-6 and TGF-β1 mRNA expression. Black seed oil treatment resulted in decreased T-cell response evident by lesser delayed type hypersensitivity and lower T-cell proliferation in spleen. In conclusion, black seed oil exhibited a significant reduction in all the markers of allergic inflammation mainly by inhibiting the delayed type hypersensitivity and T-cell proliferation. The data suggests that inhibition of T-cell response may be responsible for immunomodulatory effect of black seed oil in the rat model of allergic airway inflammation.

Establishment of a real-time PCR for quantifying transforming growth factor beta1 in blood of hepatocellular carcinoma patients

Background The carcinogenesis of hepatocellular carcinoma (HCC) is a multi-factorial, multistep and complex process. Its prognosis is poor and early detection is of the utmost importance. Transforming growth factor β1 (TGF-β1) message RNA (mRNA) has been reported to be elevated in HCC patients using Northern blotting. However, little work has been done about the detection of TGF-β1 mRNA levels in peripheral blood of patients with HCC using the real-time polymerase chain reactions (PCR) method. Objective To assess the prognostic value of quantitative levels of TGF-β1 mRNA in peripheral blood of patients with HCC, and to investigate the relationship between the expression of TGF-β1 mRNA in peripheral blood and many diagnostic and pathological factors. Methods We developed an optimized Taqman real-time PCR to quantify TGF-β1 mRNA in peripheral blood of 53 patients with HCC and 44 healthy volunteers. In addition, blood was collected from patients with HCC for measuring levels of total bilirubin (TBil), prealbumin, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyltranspeptidase (GGT), alpha-L-fucosidase (AFU), alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), viral load and platelet counts. Statistical analysis was performed using the SPSS software system (SPSS 10.0). Results In real-time PCR, fluorescence was detectable in all blood specimens from patients with HCC and healthy volunteers. The levels of TGF-β1 mRNA expression in patients with HCC were significantly higher compared to that in healthy volunteers ( P<0.000 1), suggesting an association of the activated TGF-β1 gene transcription with hepatocarcinogenesis. Patients with HCC were divided into 2 groups according to their TGF-β1 mRNA above (group A, n=28) or below (group B, n=25) the mean level. Statistical results demonstrated that TGF-β1 mRNA expression level was correlated with patients age, serum levels of CEA, CA19-9 and viral copy number ( P<0.05). Conclusion Although this is a small sample size pilot study these findings imply that quantitative measurement of TGF-β1 mRNA level in peripheral blood may be a complementary serologic marker of HCC.

Antifibrotic effects of a herbal combination regimen on hepatic fibrotic rats

Liver fibrosis has been characterized as chronic inflammatory processes involving multiple molecular pathogenetic pathways. This therapeutic study investigated whether a combination regimen of Salvia miltiorrhiza (S), Ligusticum chuanxiong (L) and Glycyrrhiza glabra (G) exerted in vivo antifibrotic effects on rats with hepatic fibrosis. Fibrosis was induced in rats by dimethylnitrosamine (DMN) administration for 4 weeks. Fibrotic rats were randomly assigned to one of the three groups: control, SLG (50 mg/kg) or silymarin (50 mg/kg), each given by gavage twice daily for 3 weeks starting 1 week after DMN injection. The results showed that fibrosis scores of livers from DMN-treated rats with SLG (1.13 +/- 0.13) were significantly reduced in comparison with DMN-treated rats receiving vehicle (1.63 +/- 0.18). Moreover, the hepatic collagen content of DMN rats was significantly reduced by either SLG or silymarin treatment. The double immunohistochemical staining results also showed that alpha-SMA positive cells with NF kappa B nuclear translocation were decreased in the fibrotic livers by SLG and silymarin treatments. The mRNA expression levels of TGF-beta1, alpha-SMA, collagen1 alpha 2, iNOS and ICAM-1 genes were attenuated by SLG and silymarin treatment. The results showed that SLG exerted antifibrotic effects in rats with DMN-induced hepatic fibrosis.

The Effect of Immunosuppression on Peritoneal Adhesions Formation After Small Bowel Transplantation in Rats

The reported incidence of adhesion related small bowel obstruction after abdominal organ transplantation is considerably lower than other abdominal procedures. The purpose of the study was to investigate the influence of immunosuppression on peritoneal adhesion formation after intestinal transplantation in rats. Four groups of rats (n = 6) underwent small bowel intestinal transplantation in syngeneic (Groups A, B) and allogeneic (Groups C, D) combinations. Groups B and D received tacrolimus immunosuppression 1 mg/kg/d. Animals were euthanized on postoperative day 7, and the total adhesion score (TAS), tissue hydroxyproline content (HPC), TGF-beta mRNA expression levels and histology were examined. All of the animals in Group C showed severe histological (Grade III) acute cellular rejection. There were no histological signs of rejection in Group D. A significant reduction in TAS was observed in tacrolimus treated animals in both syngeneic and allogeneic combinations (Groups B and D), compared with controls (Groups A and C) (P < 0.001 and P < 0.01, respectively). TAS results correlated with the differences in TGF-beta levels that showed significant reduction when each immunosuppressed group was compared with its nontreated counterpart, i.e., (Groups B versus A, P < 0.05, and Groups D versus C, P < 0.01). TGF-beta levels were significantly high in the rejection group (C) and correlated with the intense adhesion formation that was demonstrated in that group. Group C was also the only group in which a significant elevation in HPC was demonstrated (P < 0.001). Intense adhesion formation occurs during early posttransplant acute rejection. Postsurgical adhesion formation is significantly reduced in immunosuppressed rats after intestinal transplantation.

Expression of messenger RNA for transforming growth factor-beta1 and for transforming growth factor-beta receptors in peripheral blood of systemic lupus erythematosus patients treated with low doses of quinagolide

The objective of this study was to determine the expression of transforming growth factor-beta1 messenger RNA (TGF-β1 mRNA) and the expression of mRNA for TGF-β receptors (TGF-β Rs mRNA) in whole peripheral blood of consecutive (treated from several months to several years) systemic lupus erythematosus (SLE) patients (21 women). A further aim of this study was to evaluate the association between expression of the above mentioned parameters in relation to the form of applied therapy (9 patients treated with quinagolide and 12 with quinagolide plus prednisone, azathioprine or cyclosporine A). The control group consisted of 15 healthy women. Most of the patients had mild SLE with SLE disease activity index (SLEDAI) score < 10 at time when blood samples were collected. Laboratory measurements included real-time polymerase chain reaction (RT-QPCR). The expression levels of TGF-β1 mRNA and mRNA for TGF-β RII and RIII were significantly lower in patients whereas the expression level of TGF-β RI was statistically significantly higher in SLE patients than in the controls. A very high positive correlation between TGF-β1 mRNA expression and expression levels of TGF-β Rs mRNA was found. In compared subgroups selected according to the form of the applied therapy no statistically significant differences were observed. We conclude that the TGF-β signaling pathway can be altered in circulating leukocytes derived from treated patients with SLE and that the assumed forms of the applied therapy in the group of patients under consideration are accompanied by similarity in the expression level of transcripts for TGF-β1 and TGF-β Rs determined in whole blood. In our investigations, we cannot exclude the influence of the disease itself on the obtained results.

Anti-fibrotic effects of tetrandrine on bile-duct ligated rats

Tetrandrine (Tet) (C38H42O8N2; molecular weight, 622), an alkaloid isolated from the Chinese medicinal herb Stephania tetrandra, has been shown to elicit anti-inflammatory and anti-fibrotic effects in pulmonary diseases, but the mechanism of action has yet to be investigated. In this study, we tested whether Tet exerts anti-fibrotic effects on rat hepatic fibrosis through anti-NFkappaB pathways. After bile-duct ligation, rats were given Tet (1 or 5 mg/kg) or silymarin (50 mg/kg, as a positive control) by gavage twice daily for 3 weeks. Liver sections were taken for Sirius red quantitative scoring, immunofluorescence double staining of alpha-smooth muscle actin (alpha-SMA) and NFkappaB, and for quantitative determinations of the mRNA expression levels of TGF-beta1, alpha-SMA, collagen 1alpha2, inducible nitric oxide synthase (iNOS), intercellular adhesion molecule 1 (ICAM-1), metallothionein, vascular endothelial growth factor (VEGF), and VEGF type II receptor (VEGFR2) genes. The results showed that both Tet and silymarin treatment significantly reduced the fibrosis scores and hepatic collagen content of BDL rats, compared with no treatment. Both Tet and silymarin treatments decreased the number of alpha-SMA- and NFkappaB-positive cells in fibrotic livers. Moreover, Tet and silymarin treatments attenuated the mRNA expression levels of TGF-beta1,alpha-SMA, collagen 1alpha2, iNOS, ICAM-1, VEGF, and VEGFR2 genes, and induced the mRNA expression of the metallothionein gene. This study suggests that the anti-fibrotic effects of Tet were related to the reduction of fibrosis-related gene transcription, the attenuation of NFkappaB-activated pathways, and the induction of metallothionein gene transcription in the livers of BDL rats.

Up-Regulated TGF-β mRNA Expression in Splenic T Cells of High IgA-Prone Mice: A Murine Model of IgA Nephropathy with Glomerulosclerosis

Background/Aims: Recently, we established a high serum IgA-prone inbred (HIGA) mouse strain as a murine model of spontaneous IgA nephropathy by selective mating of high serum IgA ddY mice, and found that they showed enhanced production of glomerular extracellular matrix components with increased expression of TGF-β mRNA and protein in the kidneys. In this study, we examined the roles of lymphocytes in the development of high serum IgA in this strain. Methods: We performed flow cytometric analyses of T and B cells in splenic mononuclear cells (SMNCs) from these mice using BALB/c mice as normal controls. We also compared serum TGF-β1 concentrations and TGF-β mRNA expression levels in the B-cell-depleted (T-cell-rich) fraction of SMNCs in these mice. Results: HIGA mice showed significantly fewer CD3-positive cells compared with BALB/c mice when young, but not when aged. The CD4/CD8 ratio of HIGA mice was lower than that of BALB/c mice, but this difference was not significant. Although the number of B220-positive cells did not vary significantly, the ratio of surface IgA-positive B cells was significantly increased in both young and adult HIGA mice. The B-cell-depleted SMNCs from HIGA mice exhibited higher levels of expression of TGF-β and TGF-β1 mRNA than controls from a young age, which were maintained throughout life, but there were no differences in PDGF, MCP-1 or bFGF expression between these two strains. In contrast to local mRNA expression, serum TGF-β1 concentration was decreased in HIGA mice compared with BALB/c controls. Conclusion: These findings suggest that the mating procedure performed to establish HIGA mice selected for a unique phenotype of local up-regulation of TGF-β production in the kidneys, as well as T cells that may contribute to both the early and consistently high serum IgA expression and enhanced production of renal extracellular matrix components in HIGA mice. Additionally, TGF-β1 may act locally, not systemically, in a paracrine or autocrine manner.

Role of transforming growth factor beta-1 in peritonitis-induced adhesions

Peritonitis is a major cause of intra-abdominal adhesion formation. The overexpression of transforming growth factor beta-1 (TGF-β1), a potent mitogen, chemoattractant, and stimulant for collagen synthesis by fibroblasts, has been linked to tissue fibrosis at various sites throughout the body including peritoneal adhesion formation. Hence we hypothesized that the mechanism(s) involved in peritonitis-induced adhesion formation may be mediated through the upregulation of TGF-β1 expression. Peritonitis was induced in rats by cecal ligation and puncture, while a control group underwent sham operation. Adhesions were scored and harvested from both groups at 0, 6 and 12 hours and at 1, 2, 4, 7, and 28 days. Tissue expression of TGF-β1 mRNA was determined by quantitative reverse transcription—polymerase chain reaction and TGF-β1 protein was localized by immunohistochemical analysis. Serum and peritoneal fluid TGF-β1 concentrations were quantified by enzyme-linked immunosorbent assay. Compared with sham operation, peritonitis was associated with a significantly greater incidence of abdominal adhesions and a significant increase in the levels of TGF-β1 mRNA expression at days 2, 4, and 7. Immunostaining intensity of TGF-β1 in adhesions from the peritonitis group also steadily rose through day 7. In peritoneal fluid, the ratio of active:total TGF-β1 was significantly increased in the peritonitis group on days 1, 2, and 4 compared with the sham group. These results suggest that peritonitis is associated with the upregulation of TGF-β1, a mechanism that may exacerbate adhesion formation.

Differential mrna expression levels of TGFβ and LTBP-isoforms during transformation of cultured rat liver hepatic stellate cells to myofibroblasts

Differential mrna expression levels of TGFβ and LTBP-isoforms during transformation of cultured rat liver hepatic stellate cells to myofibroblasts

INHIBITION OF WOUND-INDUCED EXPRESSION OF TRANSFORMING GROWTH FACTOR-β1 mRNA BY ITS ANTISENSE OLIGONUCLEOTIDES

Transforming growth factor-β1 (TGF-β1) is a member of a gene superfamily that regulates proliferation, differentiation and other functions in many cell types. To gain insight into the role of TGF-β1 in wound repair, we have analysed the ability of an antisense TGF-β1 oligodeoxynucleotides (ODN) to specifically inhibit wound-induced expression of TGF-β1 mRNA in the mouse skin. Although injury induced the expression of TGF-β1 mRNA at the wound sites, expression of TGF-β2-or TGF-β3-mRNA was not detected. In comparison to the 24 h following injury, higher levels of TGF-β1 mRNA expression were observed in the wound sites after 72 h. Northern blotting andin situhybridisation analysis showed that wound sites treated with antisense TGF-β1 ODN exhibited no detectable levels of TGF-β1 mRNA after injury, whereas the sites treated with sense TGF-β1 ODN possessed significant amounts of its mRNA. Our results demonstrated that antisense TGF-β1 ODN inhibited the wound-induced expression of TGF-β1 mRNAin vivo.