Molecular mechanism of benign biliary stricture inhibition by rosiglitazone-activated peroxisome proliferator-activated receptor gamma

SUMMARY OBJECTIVE: The aim of this study was to investigate whether rosiglitazone-activated peroxisome proliferator-activated receptor gamma can inhibit the occurrence of benign biliary stricture and further elucidate the relevant molecular signaling mechanism. METHODS: The primary cultured rat biliary fibroblasts following experiments were performed using within the fifth generation cells, which were separated from the bile ducts of Sprague-Dawley rats. The primary cultured rat biliary fibroblasts were co-cultured with 10 ng/mL transforming growth factor-beta 1 for stimulating collagen formation. Competent cells were transfected with siRNA that specifically target Smad3 or connective tissue growth factor to inhibit the expression of the corresponding proteins. The cells were incubated with 10 μmol/L rosiglitazone to activate peroxisome proliferator-activated receptor gamma. The cells were incubated with 10 μmol/L GW9662 in the pretreatment session to inactivate peroxisome proliferator-activated receptor gamma. ELISA was used to determine the levels of connective tissue growth factor and type I collagen in the cell supernatant. Western blotting was used to detect the levels of intracellular p-Smad3/t-Smad3. RESULTS: Rosiglitazone-activated peroxisome proliferator-activated receptor gamma inhibited the secretion of type I collagen induced by transforming growth factor-beta 1. Peroxisome proliferator-activated receptor gamma inhibitor GW9662 could significantly reverse the rosiglitazone-triggered inhibition of transforming growth factor-beta 1-induced type I collagen secretion by suppressing peroxisome proliferator-activated receptor gamma activation (p<0.01). Furthermore, we also found that the activation of peroxisome proliferator-activated receptor gamma was accompanied by the inhibition of transforming growth factor-beta 1-induced Smad3 phosphorylation (p<0.01), increased connective tissue growth factor expression (p<0.01), and production of type I collagen (p<0.01), all of which effects elicited by rosiglitazone could be reversed by peroxisome proliferator-activated receptor gamma inhibitor GW9662. CONCLUSION: Peroxisome proliferator-activated receptor gamma activated by rosiglitazone inhibits the transforming growth factor-beta1 -induced phosphorylation of Smad3 and the increased connective tissue growth factor expression as well as inhibits the secretion of type I collagen in biliary fibroblasts.

Association Between Epithelial Cytokines and Clinical Phenotypes of Elderly Asthma

PURPOSE: Asthma in the elderly has different clinical features including more severe phenotypes with higher comorbidities. Epithelial cells are known to initiate innate/adaptive immune responses in asthmatic airways. We investigated clinical features and epithelial derived cytokine levels in elderly asthmatics compared to non-elderly asthmatics in a cross-sectional cohort of adult asthmatics in order to further understand its pathogenic mechanisms. METHODS: A total of 1,452 adult asthmatics were enrolled from a single tertiary hospital and were classified into 2 groups: 234 elderly (≥ 60 years at initial diagnosis) and 1,218 non-elderly (< 60 years at initial diagnosis) asthmatics. Asthma-related clinical parameters were compared between the 2 groups. Serum levels of epithelial cell-derived cytokines including interleukin (IL)-31, IL-33, IL-8, eotaxin-2, transforming growth factor beta 1 (TGF-β1) and periostin were measured by enzyme-linked immunosorbent assay. RESULTS: Significantly higher prevalence rates of late-onset asthma (onset age ≥ 40 years) and severe asthma, as well as the lower rate of atopy, blood/sputum eosinophil counts, total immunoglobulin E and eosinophil cationic protein levels were noted in elderly asthmatics compared to non-elderly asthmatics (P < 0.05, respectively). The forced expiratory volume in 1 second (FEV1, % predicted) level tended to be lower in elderly asthmatics (P = 0.07). In addition, serum IL-33 and IL-31 levels were significantly lower in elderly asthmatics, while no differences were found in the serum level of IL-8, eotaxin-2, TGF-β1 or periostin. Among elderly asthmatics, subjects with severe asthma had lower FEV1 (% predicted) value, but showed significantly higher serum levels of eotaxin-2 and TGF-β1, than those with non-severe asthma (P < 0.05 for each). CONCLUSIONS: These findings suggest that age-related changes of epithelial cell-derived cytokines may affect clinical phenotypes and severity of elderly asthma: decreased levels of IL-33 and IL-31 may contribute to less Th2 phenotype, while increased levels of eotaxin-2 and TGF-β1 may contribute to severity.

Role of the transforming growth factor (TGF)-β1 and TGF-β1 signaling pathway on the pathophysiology of respiratory pneumococcal infections / 영남의대학술지

Streptococcus pneumoniae, pneumococcus, is the most common cause of community-acquired pneumonia (CAP). CAP is an important infectious disease with high morbidity and mortality, and it is still one of the leading causes of death worldwide. Many genetic factors of the host and various environmental factors surrounding it have been studied as important determinants of the pathophysiology and outcomes of pneumococcal infections. Various cytokines, including transforming growth factor (TGF)-β1, are involved in different stages of the progression of pneumococcal infection. TGF-β1 is a cytokine that regulates a wide range of cellular and physiological functions, including immune and inflammatory responses. This cytokine has long been known as an anti-inflammatory cytokine that is critical to preventing the progression of an acute infection to a chronic condition. On the other hand, recent studies have unveiled the diverse roles of TGF-β1 on different stages of pneumococcal infections other than mitigating inflammation. This review summarizes the recent findings of the role of TGF-β1 on the pathophysiology of pneumococcal infections, which is fundamental to developing novel therapeutic strategies for such infections in immune-compromised patients.

Role of the transforming growth factor (TGF)-β1 and TGF-β1 signaling pathway on the pathophysiology of respiratory pneumococcal infections / 영남의대학술지

Streptococcus pneumoniae, pneumococcus, is the most common cause of community-acquired pneumonia (CAP). CAP is an important infectious disease with high morbidity and mortality, and it is still one of the leading causes of death worldwide. Many genetic factors of the host and various environmental factors surrounding it have been studied as important determinants of the pathophysiology and outcomes of pneumococcal infections. Various cytokines, including transforming growth factor (TGF)-β1, are involved in different stages of the progression of pneumococcal infection. TGF-β1 is a cytokine that regulates a wide range of cellular and physiological functions, including immune and inflammatory responses. This cytokine has long been known as an anti-inflammatory cytokine that is critical to preventing the progression of an acute infection to a chronic condition. On the other hand, recent studies have unveiled the diverse roles of TGF-β1 on different stages of pneumococcal infections other than mitigating inflammation. This review summarizes the recent findings of the role of TGF-β1 on the pathophysiology of pneumococcal infections, which is fundamental to developing novel therapeutic strategies for such infections in immune-compromised patients.

Diabetes mellitus affects the biomechanical function of the callus and the expression of TGF-beta1 and BMP2 in an early stage of fracture healing

Transforming growth factor beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) are important regulators of bone repair and regeneration. In this study, we examined whether TGF-β1 and BMP-2 expressions were delayed during bone healing in type 1 diabetes mellitus. Tibial fractures were created in 95 diabetic and 95 control adult male Wistar rats of 10 weeks of age. At 1, 2, 3, 4, and 5 weeks after fracture induction, five rats were sacrificed from each group. The expressions of TGF-β1 and BMP2 in the fractured tibias were measured by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction, weekly for the first 5 weeks post-fracture. Mechanical parameters (bending rigidity, torsional rigidity, destruction torque) of the healing bones were also assessed at 3, 4, and 5 weeks post-fracture, after the rats were sacrificed. The bending rigidity, torsional rigidity and destruction torque of the two groups increased continuously during the healing process. The diabetes group had lower mean values for bending rigidity, torsional rigidity and destruction torque compared with the control group (P<0.05). TGF-β1 and BMP-2 expression were significantly lower (P<0.05) in the control group than in the diabetes group at postoperative weeks 1, 2, and 3. Peak levels of TGF-β1 and BMP-2 expression were delayed by 1 week in the diabetes group compared with the control group. Our results demonstrate that there was a delayed recovery in the biomechanical function of the fractured bones in diabetic rats. This delay may be associated with a delayed expression of the growth factors TGF-β1 and BMP-2.

Transforming Growth Factor -beta 1 (TGF-beta 1) immunoreactivity in heterotopic grafts of adult dental apical papilla / Inmunoreactividad del Factor Transformador del Crecimiento-beta 1 (TGF-beta 1) en la papila apical dental adulta heterotópicamente trasplantada

To analyze the expression of transforming growth factor-beta 1 inheterotopic grafts of adult dental apical papilla. Methodology: The apical papilla of adult Wistar rats was grafted in the ear of the same donor rats. 1, 3, 7 and14 days after grafting, rats were perfused and the tissue containing the graft was processed for histological conventional technique and for immunohistochemical detection of transforming growth factor-beta 1. Results: Heterotopically grafted apical papilla developed osteoid dentine. In an early post-grafting stage, odontoblast-like cells organized themselves in palisade and synthesized dentine. However, newly formed dentine possessed the structural appearance of reactive osteoid dentine, which was systematically destroyed by the activity of osteoclaste-like cells. Transforming Growth Factor-beta 1 was observed in mesenchymal cells, extracellular matrix of the graft and surrounding host tissue, while odontoblast-like cells were systematically devoid of immunoreactivity. Conclusion: The different expression of transforming growth factor-beta 1 between normal tissue and grafted tissue development suggests that in heterotopic graft conditions the inflammatory mediation of the transforming growth factor-beta 1 prevails against its morphogenetic role...

Analizar la expresión del factor transformador del crecimiento-beta1 en trasplantes heterotópicos de papila dental del incisivo de la rata adulta. Metodología: La papila apical del incisivo de 12 ratas Wistar adultas fue trasplantada en la oreja de las mismas ratas donantes, y perfundidas 1, 3, 7 y 14 días postrasplante. El tejido fue procesado para histología convencionaly para la detección inmunohistoquímica del factor transformador del crecimiento-beta1. Resultados: La papila apical trasplantada desarrolló osteodentina. En fases tempranas postrasplante se observaron células parecidas a los odontoblastos que se organizaron en empalizada y segregaron dentina que se depositó sobre su superficie apical o secretora. Esta dentina evolucionó a osteodentina caracterizada por perder su estructura tubular e incluir a las células odontoblásticas en lagunas de su matriz. Finalmente, la osteodentina presentó procesos líticos mediados por células de tipo osteoclasto. Durante todo el proceso la expresión del factor transformador del crecimiento-beta1 se restringió a las células mesenquimales, a la matriz del trasplante y a las zonas circundantes del huésped, estando ausente en los odontoblastos, a diferencia de lo que sucede durante la odontogénesis normal. Conclusión: La diferente localización de la expresión del Factor Transformador de crecimiento beta1 entre el tejido hospedero y el trasplantado sugieren que en condiciones de trasplante heterotópico de papila dental la mediación inflamatoria del Factor Transformador de crecimiento beta1 prevalece sobre su papel morfogenético...

Cardamonin Suppresses TGF-beta1-Induced Epithelial Mesenchymal Transition via Restoring Protein Phosphatase 2A Expression

Epithelial mesenchymal transition (EMT) is the first step in metastasis and implicated in the phenotype of cancer stem cells. Therefore, understanding and controlling EMT, are essential to the prevention and cure of metastasis. In the present study, we examined, by Western blot, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy, the effects of cardamonin (CDN) on transforming growth factor-beta1 (TGF-beta1)-induced EMT of A549 lung adenocarcinoma cell lines. TGF-beta1 induced expression of N-cadherin and decreased expression of E-cadherin. CDN suppressed N-cadherin expression and restored E-cadherin expression. Further, TGF-beta1 induced migration and invasion of A549 cancer cells, which was suppressed by CDN. TGF-beta1 induced c-Jun N-terminal kinase (JNK) activation during EMT, but CDN blocked it. Protein serine/threonine phosphatase 2A (PP2A) expression in A549 cancer cells was reduced by TGF-beta1 but CDN restored it. The overall data suggested that CDN suppresses TGF-beta1-induced EMT via PP2A restoration, making it a potential new drug candidate that controls metastasis.

Inhibitory Effect of Delphinidin on Extracellular Matrix Production via the MAPK/NF-kappaB Pathway in Nasal Polyp-Derived Fibroblasts

PURPOSE: Nasal polyps are associated with chronic inflammation of the mucous membranes in the nose and paranasal sinuses and involved in extracellular matrix (ECM) accumulation. Delphinidin promotes ECM degradation in hepatitis and cardiac fibrosis. The aims of this study were to examine the inhibitory effect of delphinidin on TGF-beta1-induced myofibroblast differentiation and ECM accumulation, and to determine the underlying mechanisms in nasal polyp-derived fibroblasts (NPDFs). METHODS: NPDFs were stimulated with TGF-beta1, with or without delphinidin, and the expression levels of alpha-SMA, fibronectin, and collagen type I were determined by RT-PCR, Western blot analysis, and collagen assay. The expression of alpha-SMA protein was measured by immunocytochemical staining. Mitogen-activated protein kinase and NF-kappaB activation induced by TGF-beta1 were determined by Western blot analysis. The transcriptional activity of NF-kappaB was measured by luciferase assay. RESULTS: The expression levels of alpha-SMA, fibronectin, and collagen type I increased in TGF-beta1-stimulated NPDFs. In TGF-beta1-induced NPDFs, delphinidin inhibited the expression of alpha-SMA, fibronectin, and collagen. Inhibitors of MAPK and NF-kappaB blocked the expression of alpha-SMA, fibronectin, and collagen type I. Delphinidin suppressed the activation of MAPK and NF-kappaB induced by TGF-beta1 stimulation. CONCLUSIONS: These results suggest that delphinidin may inhibit TGF-beta1-induced myofibroblast differentiation and ECM production through the MAPK/NF-kappaB signaling pathway in NPDFs.

Lactoferrin Stimulates Mouse Macrophage to Express BAFF via Smad3 Pathway

B cell-activating factor belonging to the TNF family (BAFF) is primarily expressed by macrophages and stimulates B cell proliferation, differentiation, survival, and Ig production. In this study, we explored the effect of lactoferrin (LF) on BAFF expression by murine macrophages. We determined the level of BAFF expression at the transcriptional and protein levels using RT-PCR and ELISA, respectively. LF markedly enhanced BAFF expression in mouse macrophages at both the transcriptional and protein levels. Overexpression of Smad3/4 further increased LF-induced BAFF transcription while DN-Smad3 abolished the LF-induced BAFF expression. These results demonstrate that LF can enhance BAFF expression through Smad3/4 pathway.

Estudo imuno-histoquímico da presença de miofibroblastos e da expressão do fator transformador de crescimento-beta1, interferon gama, mataloproteinase de matriz 13 e indutor de metaloproteinases de matriz em lesões odontogênicas epiteliais / Immunohistochemical study of the presence of myofibroblasts and the expression of transforming growth factor-beta1, interferon gamma, mataloproteinase matrix 13 and matrix metalloproteinase inducer in odontogenic epithelial lesions

Os miofibroblastos são células que apresentam um fenótipo híbrido exibindo características morfológicas de fibroblastos e de células musculares lisas, sendo a aquisição de tal fenótipo denominada diferenciação, passando então a expressar a a-SMA, a qual é importante na identificação dessas células. Estudos têm sugerido que os miofibrobíastos apresentam relação com a agressividade de diversas lesões e que o seu processo de diferenciação estaria relacionado à expressão do TGF-pl e do IFN-y atuando, respectivamente, no estímulo e na inibição dessa diferenciação. O objetivo deste trabalho foi investigar o papel dos miofibroblastos em lesões odontogênicas epiteliais, relacionando-os à agressividade das lesões e analisar por meio da imuno-histoquímica. a expressão do TGF-pl e IFN-y no processo de diferenciação, além da análise da MMP-13 que é ativada por miofibroblastos e do indutor de metaloproteinases de matriz (EMMPRIN) como precursor desta MMP. A amostra foi constituída por 20 ameloblastomas sólidos, 10 ameloblastomas unicfsticos, 20 ceratocistos odontogênicos e 20 tumores odontogênícos adenomatóides. Para a avaliação dos miofibroblastos, foram quantificadas as células imunorreativas ao anticorpo a-SMA presentes no tecido conjuntivo, próximo ao tecido epitelial. As expressões de TGF-pl, IFN-y, MMP-13 e EMMPRIN, foram avaliadas no componente epitelial e no conjuntivo, estabelecendo-se o percentual de imunorreatividade e atribuindo-se escores de 0 a 4. A análise dos miofibroblastos evidenciou maior concentração nos ameloblastomas sólidos (média de 30,55), seguido pelos ceratocistos odontogênicos (22,50), ameloblastomas unicísticos (20,80) e tumores odontogênicos adenomatóides (19,15) com valor de p= 0,001. Não foi encontrada correlação significativa entre TGF-pl e IFN-y no processo de diferenciação dos miofibroblastos, bem como na relação entre a quantidade de miofibroblastos e a expressão da MMP-13. Constatou-se, correlação estatística entre MMP-13 e TGF-pi (r= 0,087; p= 0,011) além de significante correlação entre MMP-13 e IFN-y (r=0,348; p=0,003). Entre EMMPRÍN e MMP-13 verificou-se significância (r= 0,474; p<0,001) assim como entre EMMPRIN e IFN-y (r=0,393; p=0,001). A maior quantidade de miofibroblastos evidenciada nos ameloblastomas sólidos, ceratocistos odontogênicos e ameloblastomas unicísticos sugere que estas células podem ser um dos fatores responsáveis para um comportamento biológico mais agressivo destas lesões, embora a população de miofibroblastos não tenha apresentado correlação com TGF- -pi, IFN-y ,MMP-13 e EMMPRIN. Quanto a correlação evidenciada entre MMP-13 e TGF-pl, isto pode sugerir um papel indutor do TGF-pl para a expressão da MMP-13, assim como os resultados deste estudo reforçam a relação bem estabelecida do EMMPRIN como indutor da MMP-13. Constatou-se também relação entre EMMPRIN e IFN-y assim como entre MMP-13 e IFN-y sugerindo, dessa forma, um sinergismo na ação anti-fibrótica desses marcadores.

Myofibroblasts are cells that exhibit a hybrid phenotype, sharing the morphoíogical characteristics of fibroblasts and smooth muscle cells, which is acquired during a process called differentiation. These cells then start to express a-SMA, a marker that can be used for their identification. Studies suggest that myofibroblasts are related to the aggressiveness of different tumors and that TGF-pl and IFN-y play a role in myofibroblast differentiation, stimulating or inhibiting this differentiation, respectively. The objective of this study was to investigate the role of myofibroblasts in epithelial odontogenic tumors, correlating the presence of these cells with the aggressiveness of the tumor. Immunohistochemistry was used to evaluate the expression of TGF-pl and IFN-y in myofibroblast differentiation, as well as the expression of MMP-13, which is activated by myofibroblasts, and of EMMPRIN (extracellular matrix metalloproteinase inducer) as a precursor of this MMP. The sample consisted of 20 solid ameloblastomas, 10 unicystic ameloblastomas, 20 odontogenic keratocysts, and 20 adenomatoid odontogenic tumors. For evaluation of myofibroblasts, anti-a-SMA-immunoreactive cells were quantified in connective tissue close to the epithelium. Immunoexpression of TGF-pl, IFN-y, MMP-13 and EMMPRIN was evaluaíed in the epithelial and connective tissue components, attributing scores of 0 to 4. The results showed a higher concentration of myofibroblasts in solid ameloblastomas (mean of 30.55), followed by odontogenic keratocysts (22.50), unicystic ameloblastomas (20.80), and adenomatoid odontogenic tumors (19.15) (p=0.00). No significant correlation between TGF-pl and IFN-y was observed during the process of myofibroblast differentiation. There was also no correlation between the quantity of myofibroblasts and MMP-13 expression. Significant correlations were found between MMP-13 and TGF-pi (r=0.087; p=0.01 1), between MMP-13 and ÍFN-y (r=0.348; p=0.003), as well as between EMMPRIN and MMP-13 (r=0.474; /xO.001) and between EMMPRIN and IFN-y (r=0.393; p=0.00). The higher quantity of myofibroblasts observed in solid ameloblastomas, odontogenic keratocysts and unicystic ameloblastomas suggests that these cells are one of the factors responsible for the more aggressive biological behavior of these tumors, although the myofibroblast population was not correlated with TGF-01, IFN-y, MMP-13 or EMMPRIN. The correlation between MMP-13 and TGF-pl suggests that the latter induces the expression of this metalloproteinase. The present results also support the well-established role of EMMPRIN as an inducer of MMP-13. Furthermore, the relationship between EMMPRIN and IFN-y and between MMP-13 and IFN-y suggests synergism in the antifibrotic effect of these markers.

T Cell Immunoglobulin Mucin Domain (TIM)-3 Promoter Activity in a Human Mast Cell Line

T cell immunoglobulin mucin domain (TIM)-3 is an immunomodulatory molecule and upregulated in T cells by several cytokines. TIM-3 also influences mast cell function but its transcriptional regulation in mast cells has not been clarified. Therefore, we examined the transcript level and the promoter activity of TIM-3 in mast cells. The TIM-3 transcript level was assessed by real-time RT-PCR and promoter activity by luciferase reporter assay. TIM-3 mRNA levels were increased in HMC-1, a human mast cell line by TGF-beta1 stimulation but not by stimulation with interferon (IFN)-alpha, IFN-lambda, TNF-alpha, or IL-10. TIM-3 promoter -349~+144 bp region relative to the transcription start site was crucial for the basal and TGF-beta1-induced TIM-3 promoter activities in HMC-1 cells. TIM-3 promoter activity was increased by overexpression of Smad2 and Smad4, downstream molecules of TGF-beta1 signaling. Our results localize TIM-3 promoter activity to the region spanning -349 to +144 bp in resting and TGF-beta1 stimulated mast cells.

The Expressions of TGF-beta1 and IL-10 in Cultured Fibroblasts after ALA-IPL Photodynamic Treatment

BACKGROUND: Topical photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) was originally used for treating superficial skin tumors. The application of PDT to other inflammatory dermatoses like acne vulgaris, psoriasis, granuloma annulare, localized scleroderma and lichen sclerosus has recently been introduced. However, the underlying mechanisms are not well understood. We've previously reported the induction of tumor growth factor (TGF)-beta1 and interleukin (IL)-10 after PDT with ALA and intense pulsed light (IPL) in cultured HaCaT cells. OBJECTIVE: The purpose of this study was to investigate the expressions of TGF-beta1 and IL-10 in cultured fibroblasts after PDT with using ALA and IPL. METHODS: Cultured fibroblasts were treated with ALA-IPL PDT (1micromol/L of ALA; 0, 4, 8 and 12 J/cm2 of IPL). The expressions of TGF-beta1 and IL-10 were investigated by reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay. RESULTS: TGF-beta1 mRNA and protein were reduced down to 0.52- and 0.63-fold, respectively, after PDT and the IL-10 protein was increased up to 2.74-fold after PDT. CONCLUSION: The reduction of TGF-beta1 was prominent after PDT and so an antisclerotic effect can be expected after PDT. The induction of IL-10 may contribute to the anti-inflammatory effect, which explains the therapeutic benefit of PDT for inflammatory dermatoses.

Epithelial to Mesenchymal Transition of Mesothelial Cells in Tuberculous Pleurisy

PURPOSE: Tuberculous pleurisy is the most frequent extrapulmonary manifestation of tuberculosis. In spite of adequate treatment, pleural fibrosis is a common complication, but the mechanism has not been elucidated. This study is to determine whether epithelial to mesenchymal transition (EMT) of mesothelial cells occurs in tuberculous pleurisy. MATERIALS AND METHODS: Normal pleural mesothelial cells, isolated from irrigation fluids during operations for primary spontaneous pneumothorax, were characterized by immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). These cells were treated in vitro with various cytokines, which were produced in the effluents of tuberculous pleurisy. The isolated cells from the effluents of tuberculous pleurisy were analyzed by immunofluorescence and RT-PCR analysis. RESULTS: The isolated cells from the irrigation fluid of primary spontaneous pneumothorax had epithelial characteristics. These cells, with transforming growth factor-beta1 and/or interleukin-1beta treatment, underwent phenotypic transition from epithelial to mesenchymal cells, with the loss of epithelial morphology and reduction in cytokeratin and E-cadherin expression. Effluent analysis from tuberculous pleurisy using immunofluorescence and RT-PCR demonstrated two phenotypes that showed mesenchymal characteristics and both epithelial & mesencymal characteristics. CONCLUSION: Our results suggest that pleural mesothelial cells in tuberculous pleurisy have been implicated in pleural fibrosis through EMT.

The Effects of TWEAK, Fn14, and TGF-beta1 on Degeneration of Human Intervertebral Disc

OBJECTIVE: The purpose of this study is to explain the effect and reciprocal action among tumor necrosis factor (TNF) like weak inducer of apoptosis (TWEAK), fibroblast growth factor-inducible 14 (Fn14), and transforming growth factor-beta1 (TGF-beta1) on degeneration of human intervertebral disc (IVD). METHODS: Human intervertebral disc tissues and cells were cultured with Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/F-12) media in 37degrees C, 5% CO2 incubator. When IVD tissues were cultured with TWEAK, Fn14 that is an antagonistic receptor for TWEAK and TGF-beta1, the level of sulfated glycosaminoglycan (sGAG) was estimated by dimethyl methyleneblue (DMMB) assay and sex determining region Y (SRY)-box 9 (Sox9) and versican messenger ribonucleic acid (mRNA) levels were estimated by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: When human IVD tissue was cultured for nine days, the sGAG content was elevated in proportion to culture duration. The sGAG was decreased significantly by TWEAK 100 ng/mL, however, Fn14 500 ng/mL did not change the sGAG production of IVD tissue. The Fn14 increased versican and Sox9 mRNA levels decreased with TWEAK in IVD tissue TGF-beta1 20 ng/mL elevated the sGAG concentration 40% more than control. The sGAG amount decreased with TWEAK was increased with Fn14 or TGF-beta1 but the result was insignificant statistically. TGF-beta1 increased the Sox9 mRNA expression to 180% compared to control group in IVD tissue. Sox9 and versican mRNA levels decreased by TWEAK were increased with TGF-beta1 in primary cultured IVD cells, however, Fn14 did not show increasing effect on Sox9 and versican. CONCLUSION: This study suggests that TWEAK would act a role in intervertebral disc degeneration through decreasing sGAG and the mRNA level of versican and Sox9.

Expression of E-cadherin, Heat Shock Protein 47, Transforming Growth Factor beta1 and C4d in Chronic Allograft Nephropathy / 대한이식학회지

BACKGROUND: Chronic allograft nephropathy (CAN), which causes graft failure, is related to tubular atrophy and interstitial fibrosis. E-cadherin is a well-known epithelial marker and heat shock protein (HSP)-47 is a collagen-specific molecular chaperone that regulates collagen synthesis. Transforming growth factor (TGF)-beta1, a profibrotic cytokine, downregulates E-cadherin and induces expression of mesenchymal markers in an in vitro model. C4d expression is considered a poor prognostic marker for graft survival. This study evaluated the relationship between the expression of E-cadherin, HSP47, TGF-beta1, and C4d with the prognosis for CAN. METHODS: Between March 1991 and August 2007, we performed renal allograft biopsies on 42 recipients with deteriorating renal function. CAN was diagnosed according to the chronic allograft damage index (Banff classification). Renal allograft biopsies were examined for the expression of E-cadherin, HSP47, TGF-beta1, or C4d by immunohistochemistry. The HSP47, TGF-beta1, and E-cadherin staining was scored semiquantitatively by analyzing ten different fields of cortical interstitium and tubules. Biopsies with endothelial C4d staining in peri-tubular capillaries (> or =25%) were designated as C4d-positive. RESULTS: Of 42 recipients, 17 (40.5%) were in the graft survival group (GS) and 25 (59.5%) were in the graft failure group (GF). E-cadherin expression in tubular cells of the GS was much higher than that of the GF (94.1% vs 52%, P=0.04). HSP47 expression in tubular cells and interstitium in the GF was much higher than that in the GS (84% vs 35.3%, P=0.001). TGF-beta1 expression in tubular cells and interstitium in the GF was much higher than that in the GS (72% vs 23.5%, P=0.02). CONCLUSIONS: E-cadherin, HSP47, and TGF-beta1 expression was strongly correlated with the CAN prognosis.

Increased Growth Factors in Children with Mycoplasma pneumoniae Pneumonia / 소아알레르기및호흡기학회지

PURPOSE: Mycoplasma pneumoniae is a common causative agent of community acquired pneumonia in children and is well known to cause various respiratory and extrapulmonary diseases. We determined whether growth factors, including transforming growth factor (TGF)-beta1 and platelet-derived growth factor (PDGF-BB) that regulate airway fibrosis and remodeling, are increased in young children with M. pneumoniae pneumonia and also investigated if there was any difference in relation to the clinical status of the patients. METHODS: Fifty-two patients (3 to 6 years of age) who were admitted with M. pneumoniae pneumonia were enrolled and divided into 2 groups: the patients with frequent wheezing episodes (group A, n=28) and the patients with no previous history of wheeze (group B, n=24). The former group included the patients who had recurrent wheeze more than 3 times before admission. Fifteen children admitted with minor surgical problems were also studied as controls. TGF-beta1 and PDGF-BB were measured in the plasma samples collected on admission using ELISA in both patient groups and controls. RESULTS: Plasma TGF-beta1 and PDGF-BB levels were increased significantly in the patients with M. pneumoniae pneumonia as compared to the controls (P<0.01, respectively). TGF-beta1 and PDGF-BB were higher in group A than in group B, but the difference was not statistically significant (P=0.08 vs. P=0.05). In group A, TGF-beta1 was significantly higher in atopic patients than in non-atopic patients (P<0.05). CONCLUSION: Our study showed significantly increased TGF-beta1 and PDGF-BB in patients with M. pneumoniae pneumonia. It is suggested that these growth factors may play an important role in the pathogenesis of lower airway infection by M. pneumoniae.

Collagen Synthesis and Expression of TGF-beta1 in Er:glass Fractional Laser Treated Hairless Mice / 대한피부과학회지

BACKGROUND: Non-ablative dermal remodeling and ablative skin resurfacing are currently well-established skin treatment modalities. Fractional laser was recently introduced as a new concept for laser skin rejuvenation, and is characterized by creation of a dense pattern of epidermal and dermal microthermal treatment zones (MTZs). However, the precise mechanisms of dermal remodeling by Er:glass fractional laser treatment are largely unknown. OBJECTIVE: The purpose of this study was to investigate the effect of 1,550 nm Er:glass fractional laser treatment on dermal collagen synthesis and expression of TGF-beta1, a potent cytokine involved in collagen synthesis. METHODS: We treated hairless mice with several power densities (5 W 5 mJ~20 W 20 mJ), and examined the tissue samples on days 1, 30, and 90 after treatment. We analyzed the penetrating depth of laser treatment by determining dermal response through assessment of type I collagen synthesis and TGF-beta1 expression by H&E, Masson-trichrome staining, western blot analysis and immunohistochemistry staining. RESULTS: We observed, through H&E staining, that increasing the pulse energy of fractional laser treatment correlated with increasing depth of MTZ. Also, fractional laser treatment increased type 1 collagen synthesis on days 30 and 90, energy dependently. Immunohistochemical study showed that fractional laser treatment increased expression of type I collagen and TGF-beta1, energy dependently, with TGF-beta1 expression peaking on day 1. In addition, according to western blot analysis, expressions of TGF-beta1 and type I collagen were up-regulated in an energy- dependent manner. CONCLUSION: Er:glass fractional laser induced dermal remodeling by up-regulation of TGF-beta1 and type I collagen synthesis, and may be a promising modality for skin rejuvenation.

Establishment of a Radiation-Induced Fibrosis Model in BALB/c Mice / 대한방사선종양학회지

PURPOSE: Although radiation-induced fibrosis is one of the common sequelae occurring after irradiation of skin and soft tissues, the treatment methods are not well standardized. This study aimed to establish the skin fibrosis mouse model by fractionated radiation for the further mechanism studies or testing the efficacy of therapeutic candidates. MATERIALS AND METHODS: The right hind limbs of BALB/c mice received two fractions of 20 Gy using a therapeutic linear accelerator. Early skin damages were scored and tissue fibrosis was assessed by the measurement of a leg extension. Morphological changes were assessed by H&E staining and by Masson's Trichrome staining. TGF-beta1 expression from soft tissues was also detected by immunohistochemistry and PCR. RESULTS: Two fractions of 20 Gy irradiation were demonstrated as being enough to induce early skin damage effects such as erythema, mild skin dryness, dry and wet desquamation within several weeks of radiation. After 13 weeks of irradiation, the average radiation-induced leg contraction was 11.1+/-6.2 mm. Morphologic changes in irradiated skin biopsies exhibited disorganized collagen and extracellular matrix fibers, as well as the accumulation of myofibroblasts compared to the non-irradiated skin. Moreover, TGF-beta1 expression in tissue was increased by radiation. CONCLUSION: These results show that two fractions of 20 Gy irradiation can induce skin fibrosis in BALB/c mice accompanied by other common characteristics of skin damages. This animal model can be a useful tool for studying skin fibrosis induced by radiation.

Tissue Engineering of the Intervertebral Disc with Cultured Nucleus Pulposus Cells Using Atelocollagen Scaffold and Gene Therapy

STUDY DESIGN: This is an in-vitro experiment using rabbit intervertebral disc (IVD) cells and growth factors. OBJECTIVES: We wanted to determine the effect of types I, and II atelocollagen and growth factor gene therapy for matrix regeneration of rabbit IVD cells. SUMMARY OF THE LITERATURE REVIEW: Adenovirus-medicated growth factor gene therapy is efficient for matrix regeneration of the IVD. Atellocollagen has provided a favorable environment for matrix synthesis. However, a combined approach using gene and cell therapy in an atelocollagen scaffold has not yet been attempted. MATERIALS AND METHODS: Rabbit IVD cells were transduced with Ad/TGF-beta1 and Ad/BMP-2. The cells were then implanted to the atelocollagen scaffold. The [methyl-3H]thymidine incorporation for DNA synthesis and the [35S]sulfur incorporation for proteoglycan synthesis were measured. RT-PCR was performed for assessing the aggrecan, collagen type I, collagen type II and osteocalcin mRNA expressions. RESULTS: The rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type I atelocollagen showed a 130% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/TGF-beta1 and that were cultured in type II atelocollagen showed a 180% increase in new proteoglycan synthesis (p<0.05). The rabbit IVD cells with Ad/BMP-2 and that were cultured in type I atelocollagen showed a 70% increase in new proteoglycan synthesis, while the rabbit IVD cells with Ad/BMP-2 and that were cultured in type II atelocollagen showed a 95% increase (p<0.05). Rabbit IVD cells with Ad/TGF-beta1 and Ad/BMP-2 and that were cultured in type I and II atelocollagen demonstrated increased collagen type I and II mRNA expressions without an osteocalcin mRNA expression (p<0.05). CONCLUSION: Cell and gene therapy in an atelocollagen scaffold provided a efficient mechanism for chondrogenic matrix regeneration of rabbit IVD cells.

Influences of Chronic Indirect Cigarette Smoking on the Expression of TGF-beta1 and eNOS in Rat Vagina / 대한남성과학회지

PURPOSE: The objectives of this study were to evaluate the effects of chronic indirect cigarette smoking on vaginal blood flow and on histological change in a rat model. MATERIALS AND METHODS: Female Sprague-Dawley rats (12 weeks old, n=40) were devided into smoking and control group. For the exposure to passive smoking, the rat, in plastic enclosure, had a constant influx of cigarette smoke using a smoking generator for 8 weeks in smoking group. The experimental group was exposured to cigarette smoke for 1 hour, twice a day, daily for 8 weeks. Vaginal blood flow was measured by laser Doppler flowmeter. Serum estrogen concentration was measured using competitive radioimmunoassay. Immunohistochemistry and western blot analysis was done to observe the expression of TGF-beta1 and e-NOS. RESULTS: Mean vaginal blood flow (ml/min/100g tissue) significantly decreased in smoking group (13.4+/-1.6) compared to control (19.6+/-5.9)(p<0.05). The estimated concentration of serum estradiol (pg/ul) was similar between smoking (1.1+/-0.8) group and control (1.1+/-0.3) group. Vaginal histology of the cigarette smoking group was similar to the control. In the cigarette smoking group, the immunoreactivity of TGF-beta1 increased in the smooth muscle and fibroblasts. The protein expression of TGF-beta1 was increased in the smoking group (p<0.05). There was no significant differences in expression of e-NOS between two groups. CONCLUSIONS: A chronic indirect exposure to cigarette smoke significantly reduces vaginal blood flow and appears to cause vaginal tissue fibrosis in the female rat model. This suggest that cigarette smoking has adverse effects on female sexual functions and may cause sexual arousal disorder in women.