Humans ; Pulmonary Fibrosis ; metabolism ; pathology ; Signal Transduction ; physiology ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing ; physiology

DNA-Binding Proteins ; chemistry ; genetics ; physiology ; Humans ; Liver Cirrhosis ; pathology ; physiopathology ; Receptors, Transforming Growth Factor beta ; physiology ; Signal Transduction ; physiology ; Smad7 Protein ; Trans-Activators ; chemistry ; genetics ; metabolism ; physiology ; Transcription, Genetic ; physiology ; Transforming Growth Factor beta ; physiology ; Transforming Growth Factor beta1

OBJECTIVETo study the relationship between the morphologic mechanism of human embryonic epidermic cells and mesenchymal-epithelial transformation (MET) and its modulation factor.

METHODSMorphological occurrence of epidermis was detected with histologic methods in earlier period [estimated gestational age (EGA) 6-14 weeks] human embryonic skin samples. At the same time, the characteristic expression and their distribution markers of mesenchymal cells [vimentin and alpha-smooth muscle actin (alpha-SMA)], embryonic specific epidermic protein CK8&18, specific protein of epidermic stem cell CK19, transforming growth factor-beta1) (TGF-beta1) and its receptor (TGFbetaRI) in embryonic epidermis were examined with immunohistochemistry and indirect-immunofluorescent doble-labelling method.

RESULTSDuring EAG 6-8 weeks, ectodermal cells containing Vim+/alpha-SMA(-) were found to transform into epidermal stem cells with CK8&18+/CK19+. In ectodermal cells, protein expression density of TGFbetaRI was moderate (+ +), while positive signal of TGFbeta1 was weak (+/-). After EGA10 weeks, epidermal cells showed typical morphological characteristics.

CONCLUSIONSAt EGA 6-8 weeks, human embryonic skin epidermal cells began to form through MET, in which the signal pathway mediated by TGFbetaRI might play important roles, but the role of TGFbeta1 need to be further studied.

Cell Differentiation ; physiology ; Epidermis ; cytology ; embryology ; Epithelial Cells ; cytology ; Humans ; In Vitro Techniques ; Mesoderm ; cytology ; Receptors, Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; metabolism

This study was conducted to examine the effectiveness of a gene transfer of human TGF beta 1 gene into smooth muscle cells and whether the TGF beta 1 can increase elastin expression of smooth muscle cells. With the help of DOTAP, smooth muscle cells were transfected with pMAMneoTGF beta 1. The positive cell clones were selected with G418. The stable transfection and expression of TGF beta 1 in the smooth muscle cells were determined by immunofluorescence analysis. The expression of elastin in the transfected and untransfected cells were determined by in situ hybridization. The adhesion force between smooth muscle cells and matrix was detected by micropipette system. The results showed abundant TGF beta 1 stable expression in smooth muscle cells. TGF beta 1 gene can increase two-three times elastin expression and increase the adhesion between smooth muscle cells and matrix. TGF beta 1 can be used in vascular tissue engineering to increase smooth muscle cells adhesion.
Cell Adhesion ; Cells, Cultured ; Elastin ; biosynthesis ; Humans ; In Situ Hybridization ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics ; physiology ; Transforming Growth Factor beta1

OBJECTIVETo investigate the clinical significance of transforming growth factor-beta 1 (TGF-β1) in children with primary IgA nephropathy (IgAN).

METHODSThirty children who were diagnosed with primary IgAN by renal biopsy between May 2008 and October 2012 were included in the study. Thirty healthy children were used as the control group. Urinary and blood TGF-β1 levels were measured using enzyme-linked immunosorbent assay, and the protein expression of TGF-β1 in the renal tissue was measured by immunohistochemistry. The correlation between TGF-β1 levels in blood, urine, and renal tissue and their relationship with clinical indices were analyzed.

RESULTSChildren with primary IgAN had significantly higher urinary and blood TGF-β1 levels than the control group (P<0.01). Urinary TGF-β1 level was positively correlated with the pathological grade of renal tissue (r=0.557, P=0.001), and a significant positive correlation was also found between the TGF-β1 expression in the renal tissue and the pathological grade of renal tissue (r=0.682, P<0.01). There was no correlation between TGF-β1 levels in blood and renal tissue (r=0.038, P=0.844).

CONCLUSIONSUrinary TGF-β1 level is significantly positively correlated with the pathological severity of disease in children with primary IgAN. Clinical measurement of urinary TGF-β1 may be of great practical value in predicting the progression and prognosis of chronic nephropathy.

Adolescent ; Child ; Female ; Glomerulonephritis, IGA ; metabolism ; pathology ; Humans ; Kidney ; chemistry ; pathology ; Male ; Transforming Growth Factor beta1 ; analysis ; physiology ; urine

OBJECTIVE: To explore the effects of exogenous transforming growth factor-beta 1 (TGFbeta1) on peripheral nerve regeneration after the peripheral nerve injury and if TGFbeta1 regulates the expression of basic fibroblast growth factor (bFGF) in the anterior horn motoneurons of spinal cord during regeneration. METHODS: Forty-eight rats were crushed on the right sciatic nerve and then randomly divided into 2 groups: TGFbeta1 group and NS group. In TGFbeta1 group, TGFbeta1 50 microL (0.1 microg/mL) was injected into the proximal nerve near to the crushed nerve and after the operation the injured leg was injected with equal TGFbeta1 whereas the NS was replaced in the NS group. The rats of each group survived for 3, 7, 14 and 21 days after the lesion. The bFGF expression in the anterior horn motoneurons of spinal cord was detected by immunohistochemistry (IHC). Semi-thin section and Fast Blue retrograde tracing were also performed with the rats surviving for 21 days to observe the regeneration of distal end in the injured right sciatic nerve. RESULTS: The number of bFGF immunoreactive positive motoneurons in TGFbeta1 group was obviously higher than that of the NS group (P < 0.05). In the distal sciatic nerve of the rats treated with TGFbeta1, the number and diameter of regenerating myelinated axons and the thickness of myelinated sheath were more than those of the NS group (P < 0.05). The number of motoneurons in spinal cord and neurons in dorsol root ganglia (DRG) labelled with Fast Blue in the NS group was obviously lower than in the TGFbeta1 group (P < 0.01). CONCLUSION Exogenous TGFbeta1 plays an important role in promoting the peripheral nerve regeneration; TGFbeta1 up-regulates the bFGF expression in the anterior horn motoneurons of spinal cord during the peripheral nerve regeneration.
Animals ; Female ; Fibroblast Growth Factor 2 ; biosynthesis ; genetics ; Male ; Motor Neurons ; metabolism ; Nerve Regeneration ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; metabolism ; physiology ; Spinal Cord ; metabolism ; Transforming Growth Factor beta ; pharmacology ; Transforming Growth Factor beta1

AIMTo investigate the stage-specific localization of transforming growth factor (TGF) beta1 and beta3 during spermatogenesis in adult human testis.

METHODSThe localization of TGFbeta1 and beta3 was investigated by immunohistochemical staining method employing specific polyclonal antibodies.

RESULTSBoth TGFbeta1 and beta3 and their receptors were preponderant in the Leydig cells. TGFbeta1 could not be detected in the seminiferous tubules. TGFbeta3 and TGFbeta-Receptor (R) I were mainly seen in the elongated spermatids, while TGFbeta-RII in the pachytene spermatocytes and weak in the spermatogonia, spermatids and Sertoli cells. Only TGFbeta-RII was detected in the Sertoli cells. TGFbeta3, TGFbeta-RI and TGFbeta-RII showed a staining pattern dependent upon the stages of the seminiferous epithelium cycle.

CONCLUSIONTGFbeta isoforms and their receptors are present in the somatic and germ cells of the adult human testis, suggesting their involvement in the regulation of spermatogenesis.

Adult ; Humans ; Immunohistochemistry ; Leydig Cells ; metabolism ; Ligands ; Male ; Middle Aged ; Orchiectomy ; Prostatic Neoplasms ; pathology ; Receptors, Transforming Growth Factor beta ; metabolism ; Seminiferous Epithelium ; cytology ; metabolism ; Spermatids ; metabolism ; Spermatogenesis ; physiology ; Testis ; metabolism ; physiology ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; Transforming Growth Factor beta3

BACKGROUNDSMAD proteins have recently been identified as the first family of putative transforming growth factor-beta1 (TGF-beta1) signal transducers. This study was to investigate the effects of TGF-beta1 and signal protein Smad3 on rat cardiac hypertrophy.

METHODSThe incorporation of [(3)H]-leucine was measured to determine the hypertrophy of cardiomyocyte incubated with different doses of TGF-beta1 in cultured neonatal cardiomyocytes. The model of rat cardiac hypertrophy was produced with constriction of the abdominal aorta. At different times after the operation, rats were killed, and their left ventricular mass index (LVMI) determined. The mRNA expression of TGF-beta1 and Smad3 of cultured cells and hypertrophic left ventricles were assessed by RT-PCR. The protein expression of Smad3 was assessed by Western blot.

RESULTSIn cultured neonatal cardiomyocytes, TGF-beta1 significantly promoted incorporation of [(3)H]-leucine. With the concentration of 3 pg/L, it increased the expression of Smad3 in mRNA and protein levels after 15 minutes, and continued for up to 8 hours of cultured cardiomyocytes. The LVMI and the expression of TGF-beta1 (mRNA) and Smad3 (mRNA and protein) of hypertrophic left ventricle were increased by day 3 after the operation and continued to the 4th week. The peak expression of these was in the second week after operation.

CONCLUSIONTGF-beta1 has positive effects on rat cardiomyocyte hypertrophy. Signal protein Smad3 could be related to the pathologic progression of rat cardiac hypertrophy.

Animals ; Aortic Coarctation ; metabolism ; Cardiomegaly ; etiology ; Cells, Cultured ; DNA-Binding Proteins ; physiology ; Leucine ; metabolism ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Smad3 Protein ; Trans-Activators ; physiology ; Transforming Growth Factor beta ; genetics ; physiology ; Transforming Growth Factor beta1

OBJECTIVETo study the effect of β8 expression on transforming growth factor β1(TGF-β1) activation in astrocytes with oxygen glucose deprivation (OGD).

METHODSAstrocytes were cultured and then subjected to OGD to generate hypoxia-ischemia (HI) model in vitro. Immunocytochemistry was used to detect the expression and distribution of β8 in nomoxia cultured cells. β8 protein expression was quantified by Western blot at 12 hours, 1 day and 2 days after OGD. Astrocytes and luciferase reporter cells (TMLC) were co-cultured. β8 RNA interference system was established to specifically inhibit β8 expression in cultured astrocytes. TGF-β1 activation was then detected in the co-culture system.

RESULTSβ8 was mainly located in the cytoplasm and neurites of astrocytes. OGD resulted in increase of β8 protein expression at 12 hours after reoxygenation in astrocytes, which was peaked at 1 day after reoxygenation. TGF-β1 activation was in accordance with β8 expression in astrocyte-TMLC co-culture system after reoxygenation. After the inhibition of β8, TGF-β1 activation was significantly reduced in all time points.

CONCLUSIONSThe highly expressed β8 plays important roles in the regulation of TGF-β1 activation in neonatal rats with hypoxic-ischemic brain damage.

Animals ; Astrocytes ; metabolism ; Female ; Glucose ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; Integrin beta Chains ; physiology ; Male ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

BACKGROUNDTransforming growth factor beta (TGF-beta) plays an essential role in the regulation of normal physiologic processes of cells. TGF-beta has been shown to regulate several mitogen-activated protein kinases (MAPK) pathways in several epithelial cells. However, the effects of TGF-beta on soft tissue sarcoma are seldom reported. Our previous studies suggested that there should be some other signal transduction pathways besides Smads, which are important to regulate the growth of human embryonal rhabdomyosarcoma (RMS) cells. In the present study, we examined the expression and functional relations of extracellular signal-regulated kinase 2 (ERK2) and Smad4 in human RMS tissue and a RMS cell line, RD.

METHODSRD cells and normal human primary skeletal myoblasts (Mb) were treated with TGF-beta1 to establish the expression profile of ERK2 at the mRNA and protein levels detected by RT-PCR and immunofluorescence. Immunohistochemistry was used to detect the expression of ERK2 and Smad4 in 50 tissue specimens of human RMS and 23 specimens of normal skeletal muscles. Follow-up of specimens was performed 6 months to 70 months later.

RESULTSRD cells and human RMS tissues showed the higher expression of ERK2 and Smad4 than the normal control, either the protein level or the mRNA level. And, exogenous TGF-beta1 stimulation can lead to higher expression of ERK2 and its nuclear translocation, so TGF-beta1 can also activated MAPK (ERK2) pathway, resulting in a sustained activation of ERK2 for at least 2 hours. Immunohistochemistry analysis, however, showed that there was no correlation between ERK2 and Smad4 protein. The overexpression of ERK2 and Smad4 had no indicative effects on histological subtypes, histological grading, gender, age, and prognosis.

CONCLUSIONSIn RMS, signaling of TGF-beta1 from cell surface to nucleus can also be directed through the MAPK (ERK2) pathway besides the TGF-beta1/Smads pathway. The activation of ERK2 by TGF-beta1 may be Smad4 independent. Moreover, there may be some other tanglesome relationships between the TGF-beta1/Smads pathway and the MAPK pathway which takes part in the development, invasion and metastasis of tumor cells.

Cells, Cultured ; Humans ; Mitogen-Activated Protein Kinase 1 ; physiology ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; analysis ; Rhabdomyosarcoma ; metabolism ; Signal Transduction ; Smad4 Protein ; physiology ; Transforming Growth Factor beta1 ; pharmacology