OBJECTIVETo observe the influence of Niupo Zhibao pellet (NZP) on transforming growth factor-beta1 (TGF-beta1) and its receptor's expression.

METHODSEndotoxic shock model was established by intravenous injection of lipopolysaccharide (LPS) 1.5 mg/kg and intraperitoneal injection of D-galactosamine 100 mg/kg, and intervened by NZP, TGF-beta1 and its receptor's expression in lung tissue were detected by immunohistochemical method.

RESULTSNZP could enhance the TGF-beta1 and its receptor's expression in endotoxic shock lung tissue, and reduce the injury of lung.

CONCLUSIONThe mechanism of NZP in reducing endotoxic shock lung injury is possibly related with its effect in enhancing the TGF-beta1 and its receptor's expression in lung tissue.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Galactosamine ; Lipopolysaccharides ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Transforming Growth Factor beta ; biosynthesis ; Respiratory Distress Syndrome, Adult ; etiology ; metabolism ; Shock, Septic ; chemically induced ; complications ; metabolism ; Transforming Growth Factor beta ; biosynthesis ; Transforming Growth Factor beta1

To explore the relationship between inactivation of TGF-beta signaling pathway and acute leukemia, the expressions of TGF-beta1, TbetaRII and c-myc in the bone marrow mononuclear cells were detected by S-P immunocytochemical staining. The results showed that no significant difference of TGF-beta1 exepression was found between the patients and the control (P > 0.05), the expression of TbetaRII was significantly lower in patients than in control (P < 0.05) and the expression of c-myc was significantly higher in patients than in control (P < 0.05). There was no significant difference of TGF-beta1, TbetaRII and c-myc exepression between acute nonlymphoid leukemia and acute lymphoid leukemia (P > 0.05). Expressions of TbetaRII and c-myc were negatively correlated (r = -0.474, P < 0.01). In conclusion, the leukemic cells escape from the growth inhibitory effect because of the inactivation of TGF-beta signaling pathway; downregulation of TGF-beta receptor II cause c-myc overexepression and leukemogenesis.
Acute Disease ; Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Child ; Female ; Humans ; Immunohistochemistry ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-myc ; biosynthesis ; Receptors, Transforming Growth Factor beta ; biosynthesis ; Transforming Growth Factor beta1 ; biosynthesis

OBJECTIVETo investigate the influence of dermal template on the expressions of signal transduction protein Smad 3 and transforming growth factor beta1 and its receptor during wound healing process in patients with deep burns.

METHODSTwenty burn patients with excision of full thickness burn in the extremities were enrolled in the study and divided into two groups, i.e. template interfering group (E, n = 20, grafting of dermal template [allogeneic acellular dermal matrix] with razor thin autoskin) and control group (C, n = 20, grafting of razor thin autoskin only). The contralateral side served as the self-control. Tissue samples from the burn wounds were harvested at 1, 2, 3 and 4 post-operative weeks (POW) for immunohistochemistry staining. The positive expression rates of TGF-beta1, TbetaRI, TbetaRII and Smad3 proteins were determined by image analysis system.

RESULTSThe positive expressions of TGFbeta1, TbetaRI, TbetaRII and signal transduction protein Smad 3 in the tissue samples in both groups could be identified during 1 approximately 4 POW, and they diminished thereafter with the process of wound healing. The expression rate of TGF-beta1 in E group was (13.08 +/- 4.65)% at 1 POW and (9.03 +/- 1.89)% at 4 POW. The positive expression rate of above indices in E group was obviously lower than that in C group in corresponding time points (P < 0.05).

CONCLUSIONThe expression levels of TGFbeta1, TbetaRI, TbetaRII and Smad 3 protein in deep burn wounds could be lowered by mixed grafting of dermal template with razor thin autoskin, which might be beneficial in ameliorating of scar hyperplasia in the burn wound.

Adolescent ; Adult ; Burns ; metabolism ; surgery ; Dermis ; transplantation ; Humans ; Middle Aged ; Receptors, Transforming Growth Factor beta ; biosynthesis ; Signal Transduction ; Skin Transplantation ; Smad3 Protein ; biosynthesis ; Transforming Growth Factor beta1 ; biosynthesis ; Transplantation, Heterologous ; Wound Healing

OBJECTIVETo investigate the therapeutic effects of perindopril, an angiotensin-converting enzyme inhibitor, and valsartan, an angiotensin II receptor blocker on TGFbeta1 and TGFbeta receptor II mRNA, Smad3 and Smad7 on rat liver fibrosis.

METHODS60 Wistar rats were randomly divided into four groups (each group, n=15). Group 1 rats were not treated and served as healthy controls. The rats of groups 2,3,and 4 were injected with CCl(4) which induced liver fibrosis. After four weeks, group 3 rats started a treatment of perindopril, and group 4 rats with valsartan. All rats were sacrificed at the eighth week and their blood and livers were collected for analysis. The effects of perindopril and valsartan were evaluated by the levels of transforming growth factor-beta1 (TGFb1), and TGF receptor (TGFb1RII) mRNA in liver tissues by RT-PCR, the expressions and sites of TGFb1, Smad3 and Smad7 in liver tissue by immunohistochemical staining. The liver histopathology was also examined with HE staining, and the hydroxyproline in the liver and serum hyaluronic acid (HA) were examined using biochemsitry and RIA.

RESULTSCompared with the control group, the levels of TGFb1, TGFb1RII mRNA and the expression Smad3 were significantly decreased in the two treated groups, and the expression of Smad7 was also remarkably increased in the livers of rats treated with perindopril or valsartan. The histological changes of fibrosis, the hydroxyproline in the livers and HA were also improved in the treated rats.

CONCLUSIONPerindopril and valsartan have a protective effect on liver injury and can inhibit hepatic fibrosis induced by CCl(4) in rats. Their mechanisms may be associated with their effects of down-regulating TGFb1, TGFb1RII mRNA and smad3, and up-regulating Smad7 which then resulted in suppressing the activation of hepatic stellate cells.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Female ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Perindopril ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Transforming Growth Factor beta ; biosynthesis ; genetics ; Smad3 Protein ; biosynthesis ; genetics ; Smad7 Protein ; biosynthesis ; genetics ; Tetrazoles ; pharmacology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

The inhibitory mechanism of interferon-gamma (IFN-gamma) on the fibroblasts from Tenon's capsule was studied. By using immunohistochemical SP method and pathological image system, the inhibitory effects of IFN-gamma on the expression of transforming growth factor beta receptor I in the in vitro cultured fibroblasts from Tenon's capsule were quantitatively analyzed. The results showed that IFN-gamma could reduce the expression of transforming growth factor beta receptor I in the fibroblasts with the following dose-effect relationship: Y = 1937.5-134.2 Igx (r=-0.971, P<0.01). It was concluded that IFN-gamma could inhibit the expression of transforming growth factor beta receptor I in the fibroblasts from Tenon's capsule. The modulation of the transforming growth factor beta receptor I expression by IFN-gamma may be beneficial to the alleviation of the hyperplasia of scar after trabeculectomy.
Conjunctiva ; metabolism ; pathology ; Connective Tissue ; drug effects ; Fibroblasts ; metabolism ; pathology ; Filtering Surgery ; Glaucoma ; pathology ; surgery ; Humans ; Interferon-gamma ; biosynthesis ; genetics ; Receptors, Transforming Growth Factor beta ; analysis

OBJECTIVETo observe the effect of octreotide (OCT) on inhibiting hepatocellular carcinoma (HCC) and investigate its mechanisms.

METHODSNude mice bearing xenografts in situ were treated with OCT or saline control for 7 weeks since tumor implantation. The immunohistochemistry for somatostatin receptor 2 (SSTR2), cMet, transforming growth factor beta1 (TGFbeta1), phospho-Smad2, Smad4 and Smad7 was performed. SSTR2 and Smad4 mRNA expression was measured by semi-quantitative RT-PCR.

RESULTSAfter OCT treatment, the mean tumor weight in mice given OCT (0.17 +/- 0.14 g) was significantly lower than that of the control group (0.53 +/- 0.06 g). The inhibition rate of tumor was 67.9%. mRNA and protein expression of SSTR2, Smad4 in tumor cells of the treatment group were significantly more than that of the control group. cMet expression in OCT group was remarkably lower than that in control group. Between two groups, the expression of TGFbeta1, phospho-Smad2 and Smad7 were not remarkably different. In addition, phospho-Smad2 expression in HCC was significantly less than that of the normal hepatic cell.

CONCLUSIONOCT can inhibit the growth of HCC xenografts markedly. The mechanisms of OCT-induced inhibition effect may be related to up-regulating SSTR2 expression, down-regulating cMet, and recovering the function of TGFbeta/Smads-induced antitumor. In addition, the decreased expression of phospho-Smad2 may be an important feature of Bel7402 cells.

Animals ; Antineoplastic Agents, Hormonal ; therapeutic use ; Humans ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Octreotide ; therapeutic use ; Proto-Oncogene Proteins c-met ; biosynthesis ; Receptors, Somatostatin ; biosynthesis ; Smad2 Protein ; biosynthesis ; Transforming Growth Factor beta ; biosynthesis

OBJECTIVETo investigate the probabilities of core-biding factor a1 (Cbfa1) expression by human skin fibroblasts induced in vitro.

METHODSThe fibroblasts were isolated, purified from human skin, and were grown in incubation in the media of TNF-alpha, BMP-2, and combined TNF-alpha and BMP-2 at certain concentrations, respectively. The changes in biological features of these fibroblasts correlated with osteogenesis were detected by immunohistochemistry and RT-PCR assay.

RESULTSTNF-alpha could switch phenotype of collagen in fibroblasts from Type I and III to Type I and induce fibroblasts to express Ras and BMP type I receptor (BMPR-IA). TNF-alpha in combination with BMP-2 could induce fibroblasts to express Cbfa1 and osteocalcin mRNA.

CONCLUSIONHuman skin fibroblast could be induced into pro-osteoblast expressing Cbfa1, an osteoblast-specific transcription factor and a regulation of osteoblast differentiation, and combined use of TNF-alpha and BMP-2 was one of the regulating factors.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein Receptors, Type I ; Bone Morphogenetic Proteins ; pharmacology ; Cells, Cultured ; Collagen ; biosynthesis ; Core Binding Factor Alpha 1 Subunit ; Core Binding Factors ; Fibroblasts ; metabolism ; Humans ; Neoplasm Proteins ; Osteocalcin ; biosynthesis ; Protein-Serine-Threonine Kinases ; biosynthesis ; RNA, Messenger ; analysis ; Receptors, Growth Factor ; biosynthesis ; Skin ; cytology ; Transcription Factors ; biosynthesis ; genetics ; Transforming Growth Factor beta ; Tumor Necrosis Factor-alpha ; pharmacology

The present study was designed to elucidate whether the mechanism by which osthole decreases collagenI/III contents and their ratio is regulating the TGF-β/Smad signaling pathway in TGF-β1-overexpressed mouse cardiac fibroblasts (CFs). These CFs were cultured and treated with different concentrations of osthole. Our results showed that the TGF-β1 expression in the CFs transfected with that the recombinant expression plasmids pcDNA3.1(+)-TGF-β1 was significantly enhanced. After the CFs were treated with 1.25-5 μg·mL of osthole for 24 h, the mRNA and protein expression levels of collagensIand III were reduced. The collagen I/III ratio was also reduced. The mRNA and protein expression levels of TGF-β1, TβRI, Smad2/3, P-Smad2/3, Smad4, and α-SMA were decreased, whereas the expression level of Smad7 was increased. These effects suggested that osthole could inhibit collagen I and III expression and reduce their ratio via the TGF-β/Smad signaling pathway in TGF-β1 overexpressed CFs. These effects of osthole may play beneficial roles in the prevention and treatment of myocardial fibrosis.
Actins ; genetics ; Animals ; Cells, Cultured ; Collagen ; biosynthesis ; genetics ; Coumarins ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Mice ; Myocardium ; cytology ; Protein-Serine-Threonine Kinases ; genetics ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Receptor, Transforming Growth Factor-beta Type I ; Receptors, Transforming Growth Factor beta ; genetics ; Signal Transduction ; drug effects ; Smad Proteins ; genetics ; Transforming Growth Factor beta1 ; genetics

OBJECTIVETo study the expression of Smad4 and transforming growth factor-beta(1) (TGFbeta(1)), transforming growth factor-beta receptor II (TGFbetaRII) in cholangiocarcinoma tissue and its relationship with the biological behaviour and prognosis of the disease.

METHODSThe expressions of Smad4, TGFbeta(1) and TGFbetaRII were detected by immunohistochemical technique in 47 specimens of cholangiocarcinoma and the normal bile duct tissue adjacent to the tumor. The expressions of Smad4, TGFbeta(1) and TGFbetaRII were compared with the clinical stages and pathological grades of the patients.

RESULTSThe expression of TGFbeta(1) was positive in 36 cholangiocarcinomas (76.6%), which was higher than that in the normal tissue adjacent to the lesion. The positive expressions of Smad4 and TGFbetaRII were 14 (29.8%) and 28 (59.6%) in the carcinoma tissues, respectively (P < 0.05). The expression of TGFbeta(1) was related to the clinical stage, metastasis of lymph node and liver of the tumor (P < 0.05), but not with the histological grade (P > 0.05). There was positive correlation between TGFbetaRII expression and the clinical stage (P < 0.05), but no correlation between the TGFbetaRII expression and histological grade or metastasis of lymph node and liver (P > 0.05). The expression of Smad4 was associated with the histological grade, clinical stage and metastasis of lymph node and liver (P < 0.05).

CONCLUSIONSThe expressions of Smad4, TGFbeta(1) and TGFbetaRII correlate with the histological grading, clinical staging and metastasis of the lymph node and liver in cholangiocarcinoma. Combined detection of Smad4, TGFbeta(1) and TGFbetaRII may be helpful in the determination of the malignant degree and the prognosis of this disease.

Adult ; Aged ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Biomarkers, Tumor ; biosynthesis ; Cholangiocarcinoma ; metabolism ; secondary ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Protein-Serine-Threonine Kinases ; Receptors, Transforming Growth Factor beta ; biosynthesis ; Smad4 Protein ; biosynthesis ; Transforming Growth Factor beta1 ; biosynthesis

Liver cirrhosis is one of the major complications of hepatitis C virus (HCV) infection, but the mechanisms underlying HCV-related fibrogenesis are still not clear. Although the roles of HCV core protein remain poorly understood, it is supposed to play an important role in the regulation of cellular growth and hepatocarcinogenesis. The aim of this study was to examine the role of HCV core protein on the hepatic fibrogenesis. We established an in vitro co-culture system with primary hepatic stellate cell (HSC) isolated from rats, and a stable HepG2-HCV core cell line which had been transfected with HCV core gene. The expressions of fibrosis-related molecules transforming growth factor beta1 (TGF-beta1), transforming growth factor b receptor II (TGF beta RII), alpha-smooth muscle actin (alpha-SMA) and connective tissue growth factor (CTGF) were analyzed via histological or molecular methods. In addition, the expression levels of matrix metaloprotinase-2 (MMP-2) and collagen type I (Col I) from the co-cultured media were measured by zymogram and ELISA, respectively. The expressions of alpha-SMA, TGF-beta1, Col I, TGF beta RII and MMP-2 were significantly increased in the co-culture of stable HepG2-HCV core with HSC. Moreover, the significant increases of CTGF and TGF-beta1 in the HCV core-expressing cells were observed by either Northern or Western blot analysis. These results suggest that HCV core protein may contribute to the hepatic fibrogenesis via up-regulation of CTGF and TGF-beta1.
Actins/metabolism ; Animals ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Collagen Type I/metabolism ; Gelatinase A/metabolism ; Immediate-Early Proteins/*biosynthesis ; Intercellular Signaling Peptides and Proteins/*biosynthesis ; Liver/metabolism/*pathology ; Liver Cirrhosis/*metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Transforming Growth Factor beta/metabolism ; Research Support, Non-U.S. Gov't ; Transforming Growth Factor beta/*metabolism ; Up-Regulation ; Viral Core Proteins/genetics/*metabolism