• Tuberculosis and Respiratory Diseases
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• v.50 no.2
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• pp.182-195
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• 2001

Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$l_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.Background : Idiopathic pulmonary fibrosis(IPF) is a devastating illness for which there is little effective treatment. The key cytokines currently implicated in the fibrotic process are the transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), endothelin-1(ET-1) and interferon-$\gamma$(IFN-$\gamma$). The rat model for paraquat-induced pulmonary fibrosis was chosen to investigate the role of ET-1 in this disease. Both ET-1 and TGF-${\beta}_1$ expression in lung lesions were examined using immunohistochemical staining. After $Bosentan^{(R)}$ administration, an orally active ET-$1_A$ and ET-$1_B$ receptor antagonist, the degree of pulmonary fibrosis and ET-1 and TGF-${\beta}_1$ expression were analyzed. Method : Sprague-Dawley rats were divided into three groups, the control group, the fibrosis group, and the fibrosis-$Bosentan^{(R)}$-treated group. The animals were sacrificed periodically at 1, 3, 5, 7, 10, 14 days after administering saline or paraquat. The effects between groups were compared with the results of light microscopy and immunohistochemical staining for ET-1 and TGF-${\beta}_1$. The degree of fibrosis was evaluated by H&E and Masson's trichrome staining, which were graded by a computerized image analyzer. The degree of immunohistochemical staining was categorized by a semi-quantitative analysis method. Results : The lung collagen content had increased in the paraquat instillated animals by day 3, and continued to increase up to day 14. A daily treatment by gavage with $Bosentan^{(R)}$ (100mg/kg) did not prevent the increase in collagen deposition on the lung that was induced by paraquat instillation. There were increased immunohistochemical stains of ET-1 on the exudate, macrophages, vascular endothelial cells and pneumocytes in the paraquat instillated group. Furthermore, TGF-${\beta}_1$ expression was higher on the exudate, macrophages, some inflammatory cells, pneumocytes( type I, and II), vascular endothelium and the respiratory epithelial cells around the fibrotic area. After Bosentan treatment, there were no definite changes in ET-1 and TGF-${\beta}_1$ expression. Conclusion : Fibrosis of the Paraquat instillated group was more advanced when compared with the control group. In addition, there was increased ET-1 and TGF-${\beta}_1$ expression around the fibrotic area. ET-1 is associated with lung fibrosis but there was little effect of the ET-1 receptor blocker($Bosentan^{(R)}$) on antifibrosis.

• Journal of Periodontal and Implant Science
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• v.31 no.4
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• pp.787-801
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• 2001

The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely　identified the cDNA for a novel protein from cultured　PDL fibroblasts using subtraction hybridization between gingival fibroblasts and　PDL fibroblasts. The purpose　of this study was to determine the regulation by growth factors and cytokines on　PDLsl7 gene expression in　cultured human periodontal ligament cells and observe the immunohistochemical　localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

• The Journal of Internal Korean Medicine
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• v.35 no.4
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• pp.519-531
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• 2014

Objectives: Diabetic nephropathy is the most common cause of end stage renal disease. Transforming growth factor (TGF)-${\beta}1$, type IV collagen, advanced glycation end-products (AGEs), and angiotensin II type 1 receptor (AT1) are the main factors of diabetic nephropathy. We investigated the effects of Prunus on renal function and histopathological changes of diabetic nephropathy rat model induced by unilateral nephrectomy and streptozotocin. Methods: Diabetes was induced in male Sprague-Dawley rats ($290{\pm}10g$) by injecting streptozotocin (55 mg/kg) into the tail vein after unilateral nephrectomy. Rats were divided into 3 groups (n=6): normal, control, and Prunus. After 8 weeks of oral administration of Prunus extract on the Prunus group from 3 days after streptozotocin injection, we checked weight, 24 hrs urine, blood biochemistry and renal tissue to evaluate renal function and histopathological changes by examining parameters including albuminuria, BUN, creatinine, cholesterol, low density lipoprotein (LDL), triglyceride, TGF-${\beta}1$, type IV collagen, AGEs, and AT1. We also measured mRNA expression of TGF-${\beta}1$, type IV collagen, AGEs, and AT1 by Real Time polymerase chain reaction (RT-PCR). Results: Prunus decreased the amount of 24 hrs proteinuria, and inhibited histopathological changes of diabetic nephropathy including the expression and accumulation of TGF-${\beta}1$, type IV collagen and AGEs which could promote development of diabetic nephropathy. Prunus also inhibited mRNA expression of TGF-${\beta}1$, type IV collagen. Conclusions: These findings suggest that Prunus might protect the renal function and inhibit the development of renal injury by regulating factors including TGF-${\beta}1$, type IV collagen, AGEs, except AT1, so Prunus can be used for diabetic patients to prevent the progression of diabetic nephropathy.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.188-201
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• 1996

One of the initial events required for periodontal regeneration is the attachment, spreading and proliferation of fibroblasts at the healing sites. These have been reported that minocycline stimulates the attachment of gingival fibroblasts and periodontal ligament cells and $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of this study was to evaluate and confirm the effect of minocycline and $TGF-{\beta}1$ on human gingival fibroblasts and periodontal ligament cells. That gingival fibroblasts and periodontal ligament cells used in this study were obtained from the explants of healthy periodontal ligaments and gingival tissues of extracted 3rd molars or premolar teeth extracted from the patients with orthodontic treatment. The cells were cultured in ${\alpha}-MEM$(minimal essential medium) supplemented with antibiotics and FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide-95% air. Cells were used between the 5th to 8th passage in this study. The attachment and activity of both cells were evaluated by MTT assay. The results were as follows: 1. Maximum gingival fibroblast attachment was seen at a $50{\mu}g/ml$ dose of minocycline, while maximum periodontal ligament cell attachment was seen at a $100{\mu}g/ml$, and exposure of both cells to minocycline above maximal attachment dose results in a decline from maximum attachment. 2. The activity values of both cells tested minocycline were below to the control activity values at all concentrations. 3. The attachment values of both cells tested $TGF-{\beta}1$ were below or similar to control attachment values. On the above the findings, minocycline stimulated the cell attachment of gingival fibroblasts and periodontal ligament cells and $TGF-{\beta}1$ enhances the cell activity of periodontal ligament cells.

• IMMUNE NETWORK
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• v.12 no.5
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• pp.207-212
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• 2012

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-${\beta}1$ 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-${\beta}1$-induced TIM-3 promoter activities in HMC-1 cells. TIM-3 promoter activity was increased by over-expression of Smad2 and Smad4, downstream molecules of TGF-${\beta}1$ signaling. Our results localize TIM-3 promoter activity to the region spanning -349 to ＋144 bp in resting and TGF-${\beta}1$ stimulated mast cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.2
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• pp.307-313
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• 2001

Iron excess is known to affect long-term iron accumulation and tissue change such as fibrosis in liver. To determine the changes of expression level of genes associated with fibrosis by short-term iron exposure, we measured liver mRNA levels by reverse transcription polymerase chain reaction (RT-PCR) in rats fed dietary carbonyl iron (3%, wt/wt) for 9 weeks. The results showed that the expression of the collagen (I, III) and transforming growth factor (TGF)-$\beta$I mRNAs was enhanced in high-dose iron treated rat, compared to normal-dose iron treated rat. An electron microscopy study revealed that excess iron caused increase of collagen fibrils in liver. The cell shapes and compositions of hepatocytes and extracellular matrix(ECM) in liver were changed by the iron-treatment. Also, necrosised hepatocytes were broadly seen in ECM. Taken together, we suggest that iron overload affects changes of collagen and TGF-$\beta$I gene expression and these changes are associated with liver fibrogenesis.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.13
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• pp.5181-5186
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• 2014

Tumors have evolved numerous mechanisms by which they can escape from immune surveillance. One of these is to produce immunosuppressive cytokines. Transforming growth factor-${\beta}$(TGF-${\beta}$) is a pleiotropic cytokine with a crucial function in mediating immune suppression, especially in the tumor microenvironment. TGF-${\beta}$ produced by T cells has been demonstrated as an important factor for suppressing antitumor immune responses, but the role of tumor-derived TGF-${\beta}$ in this process is poorly understood. In this study, we demonstrated that knockdown of tumor-derived TGF-${\beta}$ using shRNA resulted in dramatically reduced tumor size, slowing tumor formation, prolonging survival rate of tumor-bearing mice and inhibiting metastasis. We revealed possible underlying mechanisms as reducing the number of myeloid-derived suppressor cells (MDSC) and $CD4^+Foxp3^+$ Treg cells, and consequently enhanced IFN-${\gamma}$ production by CTLs. Knockdown of tumor-derived TGF-${\beta}$ also significantly reduced the conversion of na$\ddot{i}$ve $CD4^+$ T cells into Treg cells in vitro. Finally, we found that knockdown of TGF-${\beta}$ suppressed cell migration, but did not change the proliferation and apoptosis of tumor cells in vitro. In summary, our study provided evidence that tumor-derived TGF-${\beta}$ is a critical factor for tumor progression and evasion of immune surveillance, and blocking tumor-derived TGF-${\beta}$ may serve as a potential therapeutic approach for cancer.

• Radiation Oncology Journal
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• v.32 no.3
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• pp.208-212
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• 2014

Marfan syndrome is one of the collagen vascular diseases that theoretically predisposes patients to excessive radiation-induced fibrosis yet there is minimal published literature regarding this clinical scenario. We present a patient with a history of Marfan syndrome requiring radiation for a diagnosis of a right brachial plexus malignant nerve sheath tumor. It has been suggested that plasma transforming growth factor beta 1 (TGF-${\beta}1$) can be monitored as a predictor of subsequent fibrosis in this population of high risk patients. We therefore monitored the patient's TGF-${\beta}1$ level during and after treatment. Despite maintaining stable levels of plasma TGF-${\beta}1$, our patient still developed extensive fibrosis resulting in impaired range of motion. Our case reports presents a review of the literature of patients with Marfan syndrome requiring radiation therapy and the limitations of serum markers on predicting long-term toxicity.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.2
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• pp.124-131
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• 2021

Objective: Approximately 30% of preeclamptic pregnancies exhibit abnormal liver function tests. We assessed liver injury-associated enzyme levels and circulating transforming growth factor beta (TGF-β) levels in an arginine vasopressin (AVP)-induced pregnant Sprague-Dawley rat model. Methods: Pregnant and non-pregnant Sprague-Dawley rats (n=24) received AVP (150 ng/hr) subcutaneously via mini-osmotic pumps for 18 days. Blood pressure was measured, urine samples were collected, and all animals were euthanized via isoflurane. Blood was collected to measure circulating levels of TGF-β1-3 isomers and liver injury enzymes in pregnant AVP (PAVP), pregnant saline (PS), non-pregnant AVP (NAVP), and non-pregnant saline (NS) rats. Results: The PAVP group showed significantly higher systolic and diastolic blood pressure than both saline-treated groups. The weight per pup was significantly lower in the AVP-treated group than in the saline group (p<0.05). Circulating TGF-β1-3 isomer levels were significantly higher in the PAVP rats than in the NS rats. However, similar TGF-β1 and TGF-β3 levels were noted in the PS and PAVP rats, while TGF-β2 levels were significantly higher in the PAVP rats. Circulating liver-type arginase-1 and 5'-nucleotidase levels were higher in the PAVP rats than in the saline group. Conclusion: This is the first study to demonstrate higher levels of TGF-β2, arginase, and 5'-nucleotidase activity in PAVP than in PS rats. AVP may cause vasoconstriction and increase peripheral resistance and blood pressure, thereby elevating TGF-β and inducing the preeclampsia-associated inflammatory response. Future studies should explore the mechanisms through which AVP dysregulates liver injury enzymes and TGF-β in pregnant rats.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.421-436
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• 2001

Transforming growth $factor-{\beta}(TGF-{\beta})$is a polypeptide biologic mediator considered to play a role in promoting bone formation in bony defect area. The purpose of this study was to examine the effect of $TGF-{\beta}$ to the periodontal regeneration of class III furcation defect in dogs. Classs III furcation defects were surgically created on the third and the fourth premolars bilaterally in the mandibles of eight mongrel dogs. Experimental periodontitis were induced by placing small cotton pellets into the created defects for 3 weeks. Experimental sites were divided into 4 groups according to the treatment modalities: Group I-Surgical debridement only; Group II-allogenic demineralized freeze dried bone grafting; Group III-allogenic demineralized freeze dried bone soaked in $TGF-{\beta}(4ng/10{\mu}l)$grafting; Group IV-allogenic demineralized freeze dried bone soaked in $TGF-{\beta}(20ng/10{\mu}l)$ grafting. The animals were sacrificed in the 8th week after periodontal surgery and the decalcified and undecalcified specimens were for histological and histometric examination. Although no significant differences was seen in the length of epitheial growth and connective attachment, group III showed the least apical migration among treatment groups. The amount of bone repair was significantly greater in group III, IV compared to group I and group II. New attachment formation was significantly greater in group III and group IV compared to group I and group II. These results suggest the allogenic demineralized freeze dried bone with $TGF-{\beta}$ in class III furcation defect has the potentiality of promoting alveolar bone formation and periodontal regeneration.