No abstract available.
Blotting, Western ; Cells, Cultured ; Connective Tissue Cells/drug effects/metabolism ; Endothelin-1/*biosynthesis ; Fibroblasts/*drug effects/metabolism ; Humans ; Transforming Growth Factor beta1/*pharmacology

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

Animals ; Cell Line ; Hepatic Stellate Cells ; drug effects ; metabolism ; pathology ; Plasminogen Activator Inhibitor 1 ; biosynthesis ; Rats ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo investigate the effect of interleukin-10 (IL-10) on the expression of transforming growth factor-beta(1) (TGFbeta(1)) and platelet-derived growth factor (PDGF) in hepatic stellate cells (HSC) during liver injury.

METHODSThe adenovirus vector (the titer was 1 x 10(7) efu/ml) encoded IL-10 gene was used to transfect the rat via the vein of caudal. At the same time, CCl(4) was injected into rat by a hypodermic injection. These processes went on twice a week. After eight weeks, the liver were perfused with collagenase IV and purified by density gradient centrifugation with Nycodenz for separate HSC. The level of IL-10 was measured by ELISA method; The expression of PDGF and TGFbeta(1) in HSC was detected by semi-quantitative RT-PCR and Western-blot methods.

RESULTSThe level of IL-10 in therapy group (adenovirus vector encoding IL-10 gene group) was higher than that in non-therapy group (adenovirus vector without IL-10 gene and PBS group); The expression of TGFbeta(1) mRNA, TGFbeta(1) protein and PDGF mRNA, PDGF protein in therapy group were significantly lower than that in non-therapy group (P < 0.05).

CONCLUSIONDownregulating the TGFbeta(1) and PDGF expression could be the passageway by which IL-10 alleviate the degree of proliferation and activation in hepatic stellate cells.

Animals ; Down-Regulation ; drug effects ; Genetic Therapy ; Hepatocytes ; drug effects ; physiology ; Interleukin-10 ; pharmacology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; therapy ; Male ; Platelet-Derived Growth Factor ; biosynthesis ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; drug effects ; physiology ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; Transforming Growth Factor beta1

AIMTo study the mechanisms of sodium ferulate (SF) on inhibition of collagen synthesis in hepatic stellate cells.

METHODSCollagen synthesis was analyzed by measuring 3H-proline incorporation. ELISA method was used to study the effect of SF on transforming growth factor beta1 (TGFbeta1) level in cultured HSC-T6 cell. The effect of SFon the TGFbeta1 activity in the supernatant of culture was analyzed by mink lung epithelial cell (Mv1Lu) proliferation inhibition by MTT assay.

RESULTSSF inhibited collagen synthesis in hepatic stellate cells stimulated with TGFbeta1. SF was shown to decrease TGFbeta1 level in the supernatant of HSC-T6 increased by oxidative stress. TGFbeta1 activity was intervened by SF.

CONCLUSIONSF could decrease collagen synthesis, with mechanism may be associated with that SF intervened TGFbeta1 activity, and reduced the level of TGFbeta1 increased by oxidative stress.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Coumaric Acids ; pharmacology ; Epithelial Cells ; cytology ; Hepatocytes ; cytology ; metabolism ; Lung ; cytology ; Mink ; Oxidative Stress ; Rats ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

BACKGROUND/AIMS: Tamoxifen has been tried in patients with hepatocellular carcinoma (HCC), however, its inhibitory mechanism remains unknown. In this study, we evaluated the effects of tamoxifen on HCC cell growth and the expression of transforming growth factor-beta1 (TGF-beta1) which had been known as an important cytokine in growth of HCC. METHODS: Hep 3B cells were cultivated in estrogen free media with 0.1 micro M, 0.5 micro M, 1 micro M, 5 micro M, and 10 micro M of tamoxifen for 6 days. Viable cells were counted daily and the TGF-beta1 concentrations in supernatant were measured by ELISA method. RESULTS: The number of viable HCC cells increased rather significantly after the treatment of tamoxifen of lower concentration (0.1 micro M) compared with that of the control (2.59x10(7) vs. 1.97x10(7); p<0.05). As the concentration of treated tamoxifen was higher, the number of viable HCC cells became gradually less, resulting in the significant decrease of it at the highest concentration (10 micro M) compared with that of the control (1.40x10(7) vs. 1.97x10(7); p<0.05). TGF-beta1 concentration in supernatant of tamoxifen-treated samples was significantly decreased compared with those of controls, regardless of the amount of treated tamoxifen. CONCLUSIONS: These results suggest that tamoxifen may suppress TGF-beta1 expression to an extent, although it has different effects on the proliferation of HCC cells, at the various concentrations of this agent in vitro. Such effects of tamoxifen on TGF-beta1 expression may inhibit the growth and progression of HCCs over-expressing TGF-beta1 in vivo.
Antineoplastic Agents, Hormonal/*pharmacology ; Carcinoma, Hepatocellular/*metabolism/pathology ; Cell Division/drug effects ; Cell Line, Tumor/metabolism ; Humans ; Liver Neoplasms/*metabolism/pathology ; Tamoxifen/*pharmacology ; Transforming Growth Factor beta/*biosynthesis ; Transforming Growth Factor beta1

OBJECTIVETo examine the effects of high and low concentrations of transforming growth factor (TGF) β(1) and insulin-like growth factor-I (IGF-I) on the extracelluar matrix synthesis of the self-assembled constructs of temporomandibular joint (TMJ) disc.

METHODSThe experimental groups of self-assembled constructs were exposed to IGF-I (10, 100 µg/L) and TGF-β(1) (5, 50 µg/L), the control groups were not added with any growth factors. All groups were examined at 3 and 6 weeks for gross morphological, histological, and biochemical changes. Safranin-O/fast green staining was used to examine glycosaminoglycan (GAG) distribution, picrosirius red and immunohistochemical staining to observe type I collagen distribution. Type I collagen contents were tested by ELISA assay kit, GAG contents were measured by Blyscan GAG assay kit, and the cell numbers were quantified with a Picogreen reagent kit.

RESULTSThe growth factor groups all upregulated the matrix synthesis of the self-assembled constructs compared with control groups. TGF-β(1) (5 µg/L) and IGF-I (10 µg/L) were the two most potent concentration in increasing type I collagen and GAG synthesis and cells proliferation. IGF-I group (10 µg/L) produced nearly 2 times (109.16 ± 5.12 µg) as much type I collagen as the control group (69.13 ± 5.94 µg) at 3 weeks. The matrix contents and the number of the proliferated cells in control group and all GF groups at 6 weeks were more than those at 3 weeks.

CONCLUSIONSIGF-I (10 µg/L) is the most beneficial growth factor and can be applied in tissue-engineering stratigies of the temporomandibular joint disc. At the same time, the exposure time of growth factors is another key factor that affects matrix synthesis of TMJ disc constructs.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Extracellular Matrix ; metabolism ; Glycosaminoglycans ; biosynthesis ; Goats ; Insulin-Like Growth Factor I ; pharmacology ; Temporomandibular Joint Disc ; cytology ; metabolism ; Tissue Engineering ; methods ; Transforming Growth Factor beta1 ; pharmacology

Animals ; Cells, Cultured ; Cyclosporine ; antagonists & inhibitors ; Fibronectins ; biosynthesis ; genetics ; Hepatocytes ; cytology ; drug effects ; PPAR gamma ; biosynthesis ; genetics ; Protective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta1 ; biosynthesis ; genetics

OBJECTIVETo study the expression of connective tissue growth factor (CTGF) mRNA and transforming growth factor beta 1 (TGFbeta1) mRNA in immunity-induced liver fibrosis rats and the effect of HanDanGanLe on them.

METHODSMale wistar rats were given intraperitoneal injection of porcine serum twice a week. At the beginning, the rats in HanDanGanLe-treatment group were feed with HanDanGanLe for precaution. The rats were killed after twelve weeks, then CTGF mRNA and TGFbeta1 mRNA were detected in liver samples with in situ hybridization, and the formation of liver fibrosis was observed with HE stain. The semi-quantitative

RESULTSof the two genes expression were analysed along with the stages of hepatic fibrosis.

RESULTSTypical liver fibrosis developed in the model group rats, and the positive stain of CTGF mRNA and TGFbeta1 mRNA increased, which was distributed in the areas where fibrosis occurred. There was obvious correlation between the expression strength of CTGF mRNA and TGFbeta1 mRNA (r = 0.799, P < 0.05). In the rats receiving HanDanGanLe, CTGF mRNA expression index decreased markedly (12.5+/-2.3 vs 28.8+/-1.4, t = 5.208, P < 0.01), so did TGFbeta1 mRNA expression index (25.4+/-3.2 vs 37.3+/-5.4, t = 5.655, P < 0.01). There was also significant correlation between the scores of CTGF mRNA expression and the stages of hepatic fibrosis (rs = 0.822, 0.808 in the model group and HanDanGanLe-treatment group, P < 0.05).

CONCLUSIONSThe expression of CTGF mRNA and TGFbeta1 mRNA is correlated closely with hepatic fibrosis degree. HanDanGanLe can effectively prevent the expression of CTGF and TGFbeta1. One of the mechanisms of the intervention may be its blocking the intracellular signalling pathways involved in liver fibrogenesis.

Animals ; Connective Tissue Growth Factor ; Drugs, Chinese Herbal ; pharmacology ; Immediate-Early Proteins ; biosynthesis ; genetics ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; metabolism ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1

BACKGROUNDAngiotensin II (Ang II) is a very important vasoactive peptide that acts upon hepatic stellate cells (HSCs), which are major effector cells in hepatic cirrhosis and portal hypertension. The present study was aimed to investigate the effects of Ang II and angiotensin II type 1 receptor antagonist (AT(1)RA) on the proliferation, contraction and collagen synthesis in HSCs.

METHODSHSC-T6 rat hepatic stellate cell line was studied. The proliferation of the HSC cells was evaluated by MTT colorimetric assay while HSC DNA synthesis was measured by (3)H-thymidine incorporation. The effects of angiotensin II and AT(1)RA on HSCs contraction were studied by analysis of the contraction of the collagen lattice. Cell culture media were analyzed by RT-PCR to detect secretion of collagen I (Col I), collagen III (Col III) and transforming growth factor beta1 (TGF-beta1) by enzyme linked immunosorbent assay. HSC was harvested to measure collagen I, collagen III and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA expression.

RESULTSAng II ((1 x 10(-10) - 1 x 10(-4)) mol/L) stimulated DNA synthesis and proliferation in HSCs compared with untreated control cells. AT(1)RA inhibited angiotensin II induced proliferation of HSCs. A linear increase in the contractive area of collagen lattice correlated with the concentration of angiotensin II (1 x 10(-9) - 1 x 10(-5) mol/L) and with time over 48 hours. AT(1)RA blocks angiotensin II induced contraction of collagen lattice. Col I, Col III and TGF-beta1 levels of the Ang II group were higher than those of control group and this increase was downregulated by AT(1)RA. The mRNA expressions of Col I, Col III and TIMP-1 were higher in HSCs from the Ang II group than the control group and downregulated by AT(1)RA.

CONCLUSIONSAngiotensin II increased DNA synthesis and proliferation of HSCs in a dose-dependent manner, stimulated the contraction of HSCs dose- and time-dependently. Angiotensin also promoted excretion of Col I, Col III and TGF-beta1 levels and stimulated Col I, Col III and TIMP-1 expression in HSCs. Angiotensin acts via the angiotensin II receptor because all of these effects are blocked by angiotensin II type 1 receptor antagonist.

Angiotensin II ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Calcium ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Dose-Response Relationship, Drug ; Liver ; cytology ; drug effects ; metabolism ; Rats ; Transforming Growth Factor beta1 ; biosynthesis