Role of bone marrow cells in the development of pancreatic fibrosis in a rat model of pancreatitis induced by a choline-deficient/ethionine-supplemented diet.

Bone marrow cell (BMC)-derived myofibroblast-like cells have been reported in various organs, including the pancreas. However, the contribution of these cells to pancreatic fibrosis has not been fully discussed. The present study examined the possible involvement of pancreatic stellate cells (PSCs) originating from BMCs in the development of pancreatic fibrosis in a clinically relevant rat model of acute pancreatitis induced by a choline-deficient/ethionine-supplemented (CDE) diet. BMCs from female transgenic mice ubiquitously expressing green fluorescent protein (GFP) were transplanted into lethally irradiated male rats. Once chimerism was established, acute pancreatitis was induced by a CDE diet. Chronological changes in the number of PSCs originating from the donor BMCs were examined using double immunofluorescence for GFP and markers for PSCs, such as desmin and alpha smooth muscle actin (αSMA), 1, 3 and 8 weeks after the initiation of CDE feeding. We also used immunohistochemical staining to evaluate whether the PSCs from the BMCs produce growth factors, such as platelet-derived growth factor (PDGF) and transforming growth factor (TGF) ß1. The percentage of BMC-derived activated PSCs increased significantly, peaking after 1 week of CDE treatment (accounting for 23.3±0.9% of the total population of activated PSCs) and then decreasing. These cells produced both PDGF and TGFß1 during the early stage of pancreatic fibrosis. Our results suggest that PSCs originating from BMCs contribute mainly to the early stage of pancreatic injury, at least in part, by producing growth factors in a rat CDE diet-induced pancreatitis model.

Regression of glomerulosclerosis in subtotally nephrectomized rats: effects of monotherapy with losartan, spironolactone, and their combination.

Angiotensin II accelerates and renin-angiotensin system blockade halts progression; blockade with high doses even reverses established glomerulosclerosis. Aldosterone also accelerates progression of glomerulosclerosis, partially independently of angiotensin II. The purpose of this study was to assess the relative ability of an angiotensin receptor type 1 (AT1) blocker, a mineralocorticoid receptor blocker, and their combination to reverse glomerulosclerosis. Sprague-Dawley rats were subjected to subtotal renal ablation (SNX) or sham operation. Eight weeks after surgery, they were either euthanized or allocated to treatment with vehicle, losartan, spironolactone, their combination, or unspecific antihypertensive treatment (dihydralazine) for 4 wk. Renal morphology was evaluated by stereology in tissues obtained using pressure-controlled perfusion fixation. Systolic blood pressure was significantly higher in SNX compared with sham-operated animals and decreased in all treatment groups. Compared with wk 8 after SNX, the glomerulosclerosis index (GSI) had increased further by week 12 in the vehicle- and dihydralazine-treated groups but was significantly lowered in the SNX+losartan as well as in the SNX+losartan+spironolactone groups and had not progressed further in the SNX+spironolactone group. The study confirms the partial regression of established glomerulosclerosis in subtotally nephrectomized rats after high-dose AT1 receptor blockade. Nonhyperkalemic doses of spironolactone prevented the increase but failed to decrease the GSI below the 8-wk level and preserved podocyte numbers. Combining the AT1 blocker with mineralocorticoid receptor blockade failed to further increase the regression of glomerulosclerosis.

Effects of electro-acupuncture on PDGF expression in spared dorsal root ganglion and associated dorsal horn subjected to partial dorsal root ganglionectomy in cats.

The effects of electro-acupuncture (EA) on the expression of platelet derived growth factor (PDGF) in spared dorsal root ganglion (DRG) and associated dorsal horns were evaluated in cats subjected to bilateral removal of L1-L5 and L7-S2 DRG, while sparing L6 DRG and were demonstrated using Immunohistochemistry, Western blot and RT-PCR techniques. On the acupunctured side, there was a significant increase in the total number of PDGF positive neurons. Large neurons of the L6 DRG at 7 days post operation (dpo), and small to medium-sized neurons at 14 dpo, as well as in the lamina II of the L6 spinal cord at 14 dpo was observed. The expression of PDGF protein increased significantly in the L6 DRG at 7 and 14 dpo and in the dorsal horn of the L6 spinal cord at 14 dpo while the upregulation of PDGF mRNA was seen at 3 dpo in the L6 DRG and the dorsal horn of the L3 and L6 spinal cord. These findings demonstrate that intrinsic PDGF has been upregulated in cats subjected to partial dorsal root ganglionectomy following EA, indicating endogenous PDGF is involved in promoting spinal plasticity following EA.

[Effects of Haikun Shenxi on expression of platelet-derived growth factor-B and mRNA in renal tissue of rats with adriamycin nephropathy].

OBJECTIVE: To investigate the effects of Haikun Shenxi on the expression of platelet-derived growth factor-BB (PDGF-BB) and mRNA in renal tissue of rats with adriamycin nephropathy. METHOD: Rat model was established by unilateral nephrectomy and injecting adriamycin intraperitoneally. The adriamycin-induced nephrotic rats were randomly divided into 6 groups: normal group, sham operation group, model group, lotensin treatment group, Haikun Shenxi low and high dose treatment groups (0.77, 0.08 mg x kg(-1). Ten weeks later, the 24 hour urine protein and blood biochemistry examinations and renal pathologic changes were observed, and the expression of PDGF-BB and mRNA was measured using immunohistochemical method. RESULT: Compared with model group, proteinuria and the levels of serum creatinine (Scr) , urea nitrogen (BUN) were decreased obviously in both Haikun Shenxi low and high dose groups. The expression of PDGF-BB and mRNA was mostly presented in cytoplasm of renal tubular epithelial cells and mesangial area, and it could be reduced significantly after treatment (P < 0. 05). CONCLUSION: The level of PDGF-BB and mRNA is high in renal tissue of adriamycin-induced nephrotic rats. This progress could be effectively inhibited by Haikun Shenxi and the mechanism may be that it can control the excessive expression of PDGF-BB and mRNA.

[The study of therapeutic effects of curcumin on hepatic fibrosis and variation of correlated cytokine].

OBJECTIVE: To investigate therapeutic effects of curcumin on hepatic fibrosis and the variation of correlated cytokine. METHODS: Rat models of hepatic fibrosis were made by carbon tetrachloride. Curcumin of 10, 20, 40 mg per 100 gram weight of rat were given to these rats of curcumin group respectively from ninth week. Normal, dissolvent, model and Salvia miltiorrhiza groups were made as controls. Serum levels of ALT, AST, HA, LN, PC-III were detected; Serum levels of TGF-beta1 and TNF-alpha were detected by ELISA method; Serum levels of NO were detected by chemical method. HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Grades of hepatic fibrosis were evaluated according to SSS system. Immunohistochemical staining was executed for detecting PDGF-BB in liver, and professional software for image analysis was used. RESULTS: Curcumin could decrease serum levels of ALT, AST, HA, LN, PC-III obviously, P < 0.05, which were increased in fibrotic group. Curcumin could decrease cytokine levels of NO, TGF-beta1, TNF-alpha, P < 0.05. Curcumin could obviously improve liver pathological variations of fibrotic rats. The score of hepatic fibrosis in curcumin group reduced significantly, P < 0.05. Curcumin treatment could reduce the expression of PDGF-BB, P < 0.05. These effects were dose-dependent. CONCLUSION: Curcumin can heal rat hepatic fibrosis. Effects of reducing the expression of correlated cytokines may be mechanisms of therapeutic effects of curcumin on hepatic fibrosis.

[Effects of salvianolic acid B on signal transduction induced by transforming growth factor-beta1 and platelet-derived growth factor- BB in hepatic stellate cells of rats].

OBJECTIVE: To explore the anti-hepatic fibrosis mechanisms of salvianolic acid B (SA-B). METHODS: Hepatic stellate cells (HSCs) isolated from rats were primarily cultured in uncoated plastic culture dish for 7 days, then were incubated with 10(-6) mmol/L SA-B and stimulated with 10 ng/ml transforming growth factor-beta1 (TGF-beta1) or platelet-derived growth factor-BB (PDGF-BB). Expressions of extracellular-regulated kinase (ERK) and its phosphorylation in HSC, and expressions of TGF beta1, receptor I (TbetaR I) and II (TbetaR II) and PDGF receptor beta (PDGFR-beta) on the surface of HSC induced by TGF-beta1 or PDGF-BB were detected with Western blot assay. RESULTS: SA-B inhibited the phosphorylation of ERK1/2 in HSC primary normally cultivated for 9 days stimulated or un-stimulated by TGF-beta1, but could not affect the expressions of TbetaR I and TbetaR II on the HSC surface; it down-regulated the expression of PDGFR-beta, but had no obvious effect on the phosphorylation of ERK1/2 in those HSC stimulated or un-stimulated by PDGF-BB. CONCLUSION: SA-B inhibits the ERK signal transduction induced by TGF-beta1 in HSC, which is independent of the expressions of TbetaR on HSC surface and also free from the ERK signal transduction stimulated by PDGF-BB. And its inhibition on PDGF-BB signal transduction in HSC is by way of restraining the expression of PDGFR in HSC.

[Effects of active components extracted from Qixue Bingzhi Recipe on proliferation of vascular smooth muscle cells and expressions of platelet-derived growth factor and its receptor genes].

OBJECTIVE: To observe the effects of paeoniflorin and total flavones extracted from Qixue Bingzhi Recipe on proliferation of vascular smooth muscle cells (VSMCs) cultured in endothelial cell conditioned medium (EC-CM) induced by oxidized low-density lipoprotein (ox-LDL) and the expressions of platelet-derived growth factor and its receptor genes. METHODS: The VSMCs were cultured in normal culture medium, EC-CM, simvastatin-medicated EC-CM, paeoniflorin and total flavones (low, medium and high dose) -medicated EC-CM respectively. The growth activity of VSMCs was assessed by thiazolyl blue tetrazolium bromide (MTT) assay. The expressions of platelet-derived growth factor (PDGF)-BB and platelet-derived growth factor receptor-alpha (PDGFR-alpha) mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The growth activity of VSMCs cultured in the EC-CM induced by ox-LDL was significantly higher than that in the normal culture medium (P<0.01), and the expressions of PDGF-BB and PDGFR-alpha mRNAs were obviously increased as compared with those in the normal culture medium. The growth activity of VSMCs cultured in each paeoniflorin and total flavones-medicated EC-CM was significantly lower than that in the EC-CM (P<0.01), and the expressions of PDGF-BB and PDGFR-alpha mRNAs were obviously decreased as compared with those in the EC-CM. CONCLUSION: The paeoniflorin and total flavones extracted from Qixue Bingzhi Recipe may be beneficial to the prevention and treatment of arteriosclerosis, and this efficacy may be correlated with down-regulating the expressions of PDGF-BB and PDGFR-alpha mRNAs which are related to the proliferation of VSMCs cultured in EC-CM induced by ox-LDL.

[Preventive effect of multi-glycoside of tripterygium Wilfordii Hook. f. on proteinuria and mesangial injury in experimental mesangial proliferative glomerulonephritis].

OBJECTIVE: To observe the preventive effect of multi-glycoside of Tripterygium Wilfordii Hook. f. (GYW) on proteinuria and mesentery injury in experimental mesangial proliferative glomerulonephritis in vivo. METHODS: The reversible anti-Thyl.1 antibody glomerulo nephritis model of rats was established with monoclonal antibody 1-22-3 and intervened with GTW, and a control group was set up in the same time. Changes of 24h urinary protein excretion, serum creatinine (Scr), blood urea nitrogen (BUN), total plasma protein (TP) and glomerular morphology were observed, and the level of mRNA expression of proliferative factors, including platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-beta (TGF-beta), in renal tissue was determined. RESULTS: GTW could inhibit proteinuria and mesangial injury in anti-Thyl. 1 antibody nephritis model. The PDGF-BB and TGF-beta mRNA expression in the anti-Thy1.1 antibody nephritis model rats were increased for 2.84 and 1.64 times respectively to those in the normal control group. GTW could down-regulate the over-expression of PDGF-BB mRNA by 33.1%, it was significantly different to that in the control group (P < 0.05). CONCLUSION: GTW could reduce the proteinuria and inhibit mesangial cells proliferation and extracellular matrix deposition, these effects maybe related to the down-regulating of PDGF-BB mRNA expression.

[Effect of qingfei oral liquid on protein expression of transforming growth factor-beta1 and platelet derived growth factor-BB of adenovirus type 3I, 7b induced human embryonic lung fibroblast cells].

OBJECTIVE: To explore the effect of qingfei oral liquid (QOL) contained serum on protein expression of transforming growth factor-beta1 (TGF-beta1) and platelet derived growth factor-BB (PDGF-BB) of adenovirus type 3I, 7b induced human embryonic lung fibroblast cells. METHODS: The cells were divided into 5 groups, the normal cells group (NCG), the virus control group (VCG), the blank serum group (BSG), the ribavirin group (RVG) and the QOL contained serum group (QSG). All the cells except those in the NCG were challenged by adenovirus type 3I, 7b and treated with correspondent medicine. The contents of TGF-beta1 and PDGF-BB in the supernatant of cell culture were monitored by ELISA and compared among groups. RESULTS: Contents of TGF-beta1 and PDGF-BB in VCG were significantly higher, while those in QSG were significantly lower than those in VCG (P < 0.01). CONCLUSION: Adenovirus infection can increase the protein expression of TGF-beta1 and PDGF-BB of human embryonic lung fibroblast cells. QOL can decrease the protein expression of these cytokines, which maybe one of the mechanisms of its antiviral effect.

[Protective effects of shenmai injection on the delayed injury of the cerebral neurons in rat induced by intracerebral hemorrhage].

OBJECTIVE: To observe protective effects of Shenmai (SM) injection on the delayed injury of the cerebral neurons in rat with intracerebral hemorrhage. METHOD: Rosenberg models of intracerebral hemorrhage was established and the effects of SM injection on the pathologic changes in neuronal structure, mitochondria-DNA(mtDNA)deletion, C-myc gene and expression PDGF-A gene in hippocampal CA1 areas, were investigated. RESULT: SM injection inhibited the apoptosis of pyramidal cells in the hippocampal CA1 areas, and decreased the degree of mtDNA deletion in the neurons in the injured area. SM injection had no effect on gene expression of C-myc at initial stage a intracerebral hemorrhage, but significantiy decreased the level of PDGF-A mRNA and prolonged the time of its expression. CONCLUSION: SM injection might attenuate the delayed injury induced by intracerebral hemorrhage via regulating the expression of PDGF.

The inhibitory effects of aqueous extract of Magnolia officinalis on human mesangial cell proliferation by regulation of platelet-derived growth factor-BB and transforming growth factor-beta1 expression.

Mesangial cell (MC) proliferation, mediated by platelet-derived growth factor (PDGF)-BB, transforming growth factor (TGF)-beta1, and cyclin-dependent kinases (CDK), is the common feature of glomerulosclerosis. Magnolia officinalis, stem bark of Machilus thunbergii S., has multiple pharmacological effects. In this study, we investigated the influence of aqueous extract of Magnolia officinalis on MC proliferation, DNA synthesis, and expression of PDGF-BB, TGF-beta1, CDK1, CDK2, and CDK4 in fetal bovine serum (FBS)-activated human MC. Magnolia officinalis inhibited the MC proliferation, DNA synthesis, and the expression of PDGF-BB, CDK1, and CDK2 gene and CDK1, CDK2, and TGF-beta1 protein. These results suggest that the inhibitory effect of Magnolia officinalis on MC proliferation may be mediated by regulation of PDGF-BB and TGF-beta1expressions and by modulation of CDK1 and CDK2 expression.

Basic studies on the clinical applications of platelet-rich plasma.

Platelets, which contain many growth factors such as platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta), are being used in clinical applications as platelet-rich plasma (PRP). Only a few studies, however, have been conducted on the growth factors present in PRP and on the clinical applications using the drug delivery system (DDS). For the purpose of clinical application, we first modified the PRP preparation method and assessed the amounts of growth factors contained in the human platelet concentrates. Furthermore, we assessed fibrin glue as a DDS of platelet concentrates. Platelet precipitations were made by twice centrifuging human whole blood. The precipitated platelet was resuspended to yield the platelet concentrates. The growth factor concentrations were measured. Fibrin glue sheets containing this platelet concentrate were implanted in rabbit pinna and samples were obtained for immunostaining (anti-PDGF antibody) to assess the use of PRP over time using the fibrin glue as the DDS. The mean concentration of growth factors present in the platelet concentrates was three times or greater than that of conventional PRP. Furthermore, the results indicated that when the platelet concentrate was used with fibrin glue as a carrier, the contents were released over a period of about 1 week. This raises the possibility that this system may be useful in clinical applications.

Platelet-derived growth factor-producing cells immortalized from rat mesencephalon with SV40 large T antigen transduced by an AAV vector.

PURPOSE: Adeno-associated virus (AAV) can infect a wide variety of mammalian cell types and is capable of infecting both dividing and non-dividing cell populations. Here we report the construction of a recombinant AAV vector which expresses the SV40 large T protein (AAV-T) and the use of this vector to immortalize primary cells from embryonic rat mesencephalon. METHODS: The AAV-T vector was constructed by introducing the BamH1 fragment of the pCMV/SVE/Neo plasmid containing T antigen and SV40 regulatory elements into the JM48 plasmid containing the inverted terminal repeats of AAV. Neuronal cultures from E-12 rat mesencephalon were grown in defined media supplemented with basic fibroblast growth factor. These cells were infected with the AAV-T vector. RESULTS: A cell line (designated RMAT) and six subclones were established from these cultures through multiple passages. This cell line was immunoreactive for SV40 large T antigen and the cytoskeletal proteins nestin and vimentin. Morphological differentiation and expression of neurofilament 160 kDa were induced by exposure to dibutyrl cyclic AMP. Immunoassays performed to measure endogenous production of growth factors showed that RMAT cells produced high levels of platelet-derived growth factor (PDGF). CONCLUSIONS: AAV may be a useful vector for the transduction of oncogenes to produce cell lines.

[The expression of platelet-derived endothelial cell growth factor in liver cancer].

OBJECTIVE: To investigate the expression of platelet-derived endothelial cell growth factor (PD-ECGF) in liver cancer and explore its related intervention with Capecitabine for the growth and metastasis of liver cancer. METHODS: The protein level of PD-ECGF was determined using immunohistochemical method in 61 HCC samples and the mRNA level was detected using Northern blot analysis. Capecitabine was administered orally in 24 nude mice bearing LCI-D20 tumor. All treatments lasted 3 weeks. The tumor size was calculated by the following formula: V = a x b2 x 0.5. Lung metastasis was evaluated by HE staining in lung samples. RESULTS: The PD-ECGF expression rate in 61 HCC and paratumoral liver tissues was 70.5% and 47.5%, respectively. The rate was higher in HCC tissues with advanced TNM stage than in those with early TNM stage (81.1% vs 54.2%, P < 0.05). The mRNA level of PD-ECGF was related well to its protein expression. The tumor size on day 3 after last treatment measured in the control, and low dose (1.05 mmol.kg-1.day-1), moderate dose (2.10 mmol.kg-1.day-1) and high dose (3.15 mmol.kg-1.day-1) of Capecitabine treatment groups was 447 mm3 +/- 159 mm3, 414 mm3 +/- 97 mm3, 240 mm3 +/- 119 mm3 and 209 mm3 +/- 150 mm3, respectively. High doses of Capecitabine increased the inhibitory effect on the growth and lung metastasis of HCC implant. CONCLUSION: PD-ECGF is highly expressed in HCC and correlates with the TNM staging. Capecitabine may inhibit the growth and metastasis of HCC.

[Effects of shuanglong pill on mRNA expression of platelet-derived growth factor A and monocyte chemoattractant protein-1 mRNA].

OBJECTIVE: To explore the molecular mechanism of Shuanglong Pill (SLP) in intervening atherosclerosis, mainly the effect of SLP on the mRNA expression of the platelet-derived growth factor-A (PDGF-A) and the monocyte chemoattractant protein-1 (MCP-1). METHODS: Using the hyperlipoprotein serum (HLS) of cultured rabbits' aortic smooth muscle cell (SMC) to build the atherosclerotic cell model, and medicated serum acting on the cell model by serum pharmacologic method. The effect of SLP medicated serum on the level of PDGF-A, MCP-1 mRNA expression were observed and analysed by means of reverse transcriptase-polymerase chain reaction assay. RESULTS: It was found that HLS stimulated mRNA expression of the PDGF-A and the MCP-1, but the medicated serum of SLP lowered their expression level. CONCLUSION: SLP could inhibit the function of cellular auto-secreting growth factor, it is one of the possible pharmacologic mechanism in intervening atherosclerosis.

Vascular smooth muscle cell proliferation and regrowth after mechanical injury in vitro are Egr-1/NGFI-A-dependent.

Smooth muscle cell (SMC) proliferation is a key event in renarrowing of blood vessels after balloon angioplasty. Mechanical injury imparted to the arterial wall in experimental models induces the expression of the immediate-early gene, egr-1. Egr-1 binds to and activates expression from the proximal promoters of multiple genes whose products can, in turn, influence the vascular response to injury. Here, we used antisense strategies in vitro to inhibit rat vascular SMC proliferation by directly targeting Egr-1. A series of phosphorothioate antisense oligonucleotides of 15 base length and complementary to various theoretically accessible regions within Egr-1 mRNA were synthesized and assessed for their ability to selectively inhibit SMC proliferation in an Egr-1-dependent manner. Western blot analysis revealed that two oligonucleotides, AS2 and E11, inhibited Egr-1 synthesis in cells exposed to serum without affecting levels of the zinc finger protein Sp1. AS2 and E11 inhibited serum-inducible [(3)H]thymidine incorporation into DNA, as well as serum stimulation of total cell numbers. Size-matched phosphorothioate oligonucleotides with random, scrambled, sense or mismatch sequences failed to inhibit. Antisense Egr-1 inhibition was nontoxic and reversible. These oligonucleotides also inhibited SMC regrowth after mechanical injury in vitro. Egr-1 thus plays a key regulatory role in SMC proliferation and repair following injury.

Effects of immunosuppressants on platelet-derived growth factor-A chain mRNA expression and coronary arteriosclerosis in rat cardiac allografts.

Graft coronary arteriosclerosis (GCA) that results in proliferative and obstructive lesions limits the long-term success of cardiac transplantation. Despite extensive study, the pathogenic mechanisms underlying GCA are still unclear and therapeutic strategies for this condition have been inadequate. In this study, we compared the therapeutic effectiveness of cyclosporine A (CsA), 15-deoxyspergualin (DSG), and Multiglycosidorum tripterygii (MT) on GCA. In addition, we studied the correlation between the extent of GCA and the degree of platelet-derived growth facter (PDGF)-A chain mRNA expression in cardiac grafts. Lewis rats receiving heterotropic heart transplants from Wistar King donors were treated with 10 mg kg(-1) day(-1) of CsA (n=7), 5 mg kg(-1) day(-1) of DSG (n=7) or 30 mg kg(-1) day(-1) of MT (n=7) respectively. Histological evaluation of coronary arteriosclerosis and Northern blot analysis of cardiac allograft PDGF-A chain mRNA expression were conducted on day 60 after transplantation. Varying levels of GCA were observed in the 21 transplanted hearts. Significant differences in both the degree of PDGF-A mRNA expression and the extent of GCA were found among the 3 groups. GCA was significantly reduced in allografts treated with MT or DSG in comparison with the level seen in CsA-treated grafts. A significant correlation was found between PDGF-A chain mRNA expression and the grade of arterial intimal thickening (r=0.76, p<0.05) as well as with the incidence of diseased vessels (r=0.82, p<0.01). Our results indicate that both MT and DSG are more effective in the treatment of GCA than CsA. In our cardiac allografts, the degree of PDGF-A chain mRNA expression correlated well with the extent of GCA, suggesting that PDGF-A may play an important role in the development of transplant-related GCA.

[Effect of Tripterine on mRNA expression of c-myc and platelet derived growth factor of vascular smooth muscle cell in rats].

OBJECTIVE: To observe the effect of Tripterine on mRNA expression of c-myc and platelet derived growth factor (PDGF) in vascular smooth muscle cell (VSMC) of rats. METHODS: 5-10th passage culture of VSMC was used. Tripterine and 20% fetal calf serum were added into the medium of cultured VSMC in concentration of 0.1, 0.2 and 0.3 mg/L after free-serum cultivation for 24 hours. The general RNA was isolated from VSMC at 6 and 12 hours after the drug addition for detection of mRNA expression of c-myc and PDGF respectively by dot blot hybridization. The cDNA probes were labeled by digoxin. RESULTS: Tripterine inhibited the PDGF mRNA expression of VSMC and decreased c-myc mRNA expression in a dose-dependent manner, either vs. control. CONCLUSION: Tripterine could inhibit mRNA expression of c-myc and PDGF in VSMC, therefore, it would inhibit overproliferation of VSMC.

Temporal and spatial immunolocalization of cytokines in murine chronic granulomatous tissue. Implications for their role in tissue development and repair processes.

BACKGROUND: Cytokines have profound effects on various aspects of granulomatous tissue formation. However, there is little information regarding their distribution during tissue development. This study investigated the temporal and spatial distribution of transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1) and IL-1 beta in developing granulomatous tissue. EXPERIMENTAL DESIGN: Murine chronic granulomatous air pouches were induced and full thickness biopsies taken at intervals up to 28 days. Samples were prepared for immunohistochemistry and labeled using antibodies against TGF-beta, bFGF, PDGF, EGF, TNF-alpha, IL-1 alpha and IL-1 beta. RESULTS: Immunoreactivity to TGF-beta, PDGF, TNF-alpha, IL-1 alpha and IL-1 beta was localized to a proportion of macrophages within the granulomatous tissue. Immunopositive macrophage numbers increased with time, and with the exception of PDGF were associated with areas of fibrogenesis between days 14 to 28. Heterogeneous labeling of capillaries for EGF was observed within the granulomatous tissue juxtaposed to dermal musculature. Diffuse labeling of bFGF, associated with extracellular matrix, was always observed. After day 14, bFGF immunoreactivity was discretely localized to endothelial cells and the basement membrane of vessels within the granulomatous tissue. TGF-beta immunoreactivity was also associated with extracellular matrix components, being most intense in the area of fibrogenesis between 14 and 28 days. Occasional fibroblasts were also labeled with TGF-beta in this region. CONCLUSIONS: The spatial and temporal confinement of the individual cytokines suggests that a sequential coordinated process of repair and fibrosis is occurring. It is hoped that these observations will provide a more effective therapeutic approach for the sequential application of cytokines in abnormalities of wound healing.