• BMB Reports
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• v.55 no.4
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• pp.192-197
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• 2022

Cell signals for growth factors depend on the mechanical properties of the extracellular matrix (ECM) surrounding the cells. Microtubule acetylation is involved in the transforming growth factor (TGF)-β-induced myofibroblast differentiation in the soft ECM. However, the mechanism of activation of α-tubulin acetyltransferase 1 (α-TAT1), a major α-tubulin acetyltransferase, in the soft ECM is not well defined. Here, we found that casein kinase 2 (CK2) is required for the TGF-β-induced activation of α-TAT1 that promotes microtubule acetylation in the soft matrix. Genetic mutation and pharmacological inhibition of CK2 catalytic activity specifically reduced microtubule acetylation in the cells cultured on a soft matrix rather than those cultured on a stiff matrix. Immunoprecipitation analysis showed that CK2α, a catalytic subunit of CK2, directly bound to the C-terminal domain of α-TAT1, and this interaction was more prominent in the cells cultured on the soft matrix. Moreover, the substitution of alanine with serine, the 236th amino acid located at the C-terminus, which contains the CK2-binding site of α-TAT1, significantly abrogated the TGF-β-induced microtubule acetylation in the soft matrix, indicating that the successful binding of CK2 and the C-terminus of α-TAT1 led to the phosphorylation of serine at the 236th position of amino acids in α-TAT1 and regulation of its catalytic activity. Taken together, our findings provide novel insights into the molecular mechanisms underlying the TGF-β-induced activation of α-TAT1 in a soft matrix.

• The Korea Journal of Herbology
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• v.39 no.4
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• pp.19-28
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• 2024

Objectives : Ephedrae Herba has been used in the East Asian traditional medicine, for treatment of asthma, cold and influenza. Silymarin is an effective antioxidant and its anti-fibrogenic, anti-inflammatory, and hepatoprotective properties have been reported. This study was performed to explore an anti-fibrogenic potential of Ephedrae Herba extract (EHE) + silymarin on immortalized human hepatic stellate cell line, LX-2 cells. Methods : We studied the anti-fibrogenic effects of EHE + silymarin on transforming growth factor β1 (TGF-β1) signaling pathway in LX-2 cells. Cell viability was measured using the MTT assay. mRNA levels were detected by real-time PCR. TGF-β1 signaling-related proteins expression were detected by Western blot. Results : Silymarin 30 ㎍/mL and EHE 100 ㎍/mL showed cytotoxicity on LX-2 cells. Therefore, the concentrations of silymarin and EHE were studied at 10 ㎍/mL, respectively. Silymarin significantly reduced PAI-1 protein expression, Smad binding element (SBE) luciferase activity, and mRNA (PAI-1, MMP2 and 9) expression compared to TGF-β1. EHE significantly reduced SBE luciferase activity and mRNA (PAI-1, MMP2 and 9) expression compared to TGF-β1. More importantly, EHE + silymarin significantly reduced all parameters compared to TGF-β1, and also significantly reduced compared to EHE alone and silymarin alone. Conclusion : The results indicate that EHE + silymarin has anti-fibrogenic effect in LX-2 cells induced by TGF-β1. Additionally, EHE + silymarin shows more effective anti-fibrogenic effect than EHE alone and silymarin alone.

• IMMUNE NETWORK
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• v.23 no.5
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• pp.37.1-37.11
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• 2023

Forkhead box P3-positive (Foxp3⁺)-inducible Tregs (iTregs) are readily generated by TGF-β1 at low TCR signaling intensity. TGF-β1-mediated Foxp3 expression is further enhanced by retinoic acid (RA) and lactoferrin (LF). However, the intensity of TCR signaling required for induction of Foxp3 expression by TGF-β1 in combination with RA and LF is unknown. Here, we found that either RA or LF alone decreased TGF-β1-mediated Foxp3 expression at low TCR signaling intensity. In contrast, at high TCR signaling intensity, the addition of either RA or LF strongly increased TGF-β1-mediated Foxp3 expression. Moreover, decreased CD28 stimulation was more favorable for TGF-β1/LF-mediated Foxp3 expression. Lastly, we found that at high signaling intensities of both TCR and CD28, combined treatment with TGF-β1, RA, and LF induced robust expression of Foxp3, in parallel with powerful suppressive activity against responder T cell proliferation. Our findings that TGFβ/RA/LF strongly generate high affinity Ag-specific iTreg population would be useful for the control of unwanted hypersensitive immune reactions such as various autoimmune diseases.

• Korean Journal of Organic Agriculture
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• v.26 no.1
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• pp.151-161
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• 2018

This study was designed to investigate the effects of multi-enzyme on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.1% multienzyme (Multi; mixture of ${\beta}-mannanase$, xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Frequency of diarrhea, levels of packed cell volume (PCV), white blood cells (WBC), immunoglobulins, cortisol, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), and C-reactive protein (CRP) were measured. Multi group tended to decrease (p<0.1) diarrhea frequency than CON group during 2 wk after weaning. Lower values of PCV on d 3 (p<0.05) and d 7 (p<0.1) were found in Multi group compared with CON group. There were no significant differences on WBC number and immunoglobulin (Ig) M and A between Multi and CON groups. However, Multi group tended to increase (p<0.1) Ig G on d 7 than CON group. Moreover, Multi group showed modulated immune responses, indicated by decreased levels of cortisol (p<0.05) on d 7 and 14, $TNF-{\alpha}$ on d 3 (p<0.05) and d 7 (p<0.10), $TGF-{\beta}$ on d 2 (p<0.05) and d 7 (p<0.10), and CRP (p<0.10) on d 3 and 7 after weaning compared with CON group. Consequently, inclusion of multi-enzyme in diets for weaned pigs improved gut health and modulated immune responses of weaned pigs.

• Herbal Formula Science
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• v.32 no.2
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• pp.141-153
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• 2024

Objective : Ephedrae Herba (Ephedra) has been frequently used in the East Asian traditional medicine including Korea, China and Japan in the clinical treatment of asthma, cold and influenza etc. This study was performed to explore an anti-fibrogenic potential of Ephedra Herba water extract (EHE) using immortalized human hepatic stellate cell line, LX-2 cells. Methods : We examined the anti-fibrogenic effects of EHE on canonical pathway of transforming growth factor-β1 (TGF-β1) signaling in LX-2 cells. Cell viability was measured using the MTT assay. mRNA levels were detected by real-time PCR. Proteins expression were detected by Western blot. Results : Treatment of EHE 30 ㎍/ml did not show any cytotoxicity on LX-2 cells. Pre-treatment of EHE (30 ㎍/mL) significantly inhibited α-smooth muscle actin expression induced by TGF-β1. Additionally, EHE significantly decreased Smad2 and Smad3 phosphorylations, Smad binding element-driven luciferase activity and plasminogen activator inhibitor type 1 expression by TGF-β1. Furthermore, increases of matrix metalloproteinases 2 genes by TGF-β1 was also attenuated by EHE treatment. Conclusion : These results suggest that EHE has an ability to suppress fibrogenic process in activated HSC via inhibition of TGF-β1-TGFBR mediated canonical (Smad dependent) pathway.

• Korean Journal of Psychosomatic Medicine
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• v.28 no.2
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• pp.177-184
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• 2020

Objectives : Postpartum depression is known to occur in 10-15% of mothers. The concentration of cytokine varies depending on stress, depression, pregnancy and general medical conditions. We hypothesized that the concentration of cytokines may be related to reproduction and childbirth, and that women with postpartum depression would show alterations in cytokines levels. Methods : A total of 104 pregnant women were selected as subjects, and 60 non-pregnant women were selected as normal controls. Symptoms of depression were evaluated in the pregnant study subjects using the diagnostic criteria outlined in the Edinburgh Postnatal Depression Scale (EPDS). The pregnant subjects were divided into three groups perinatal non-depression controls (n=61), postpartum depression-recovery (n=18), and postpartum depression (n=25). Results : The plasma concentration of TGF-β1, IGF-1 was higher in the pregnant group than in non-pregnant controls (TGF-β1 ; p<0.01, IGF-1 ; p=0.026). At 24 weeks of pregnancy and 6 weeks of delivery, there were no significant differences in the plasma concentration of TGF-β1, IGF-1, β-NGF, IL-2, IL-4, IL-6, IFN-γ, TNF-α between the three groups. There was no statistically significant difference in all three groups during the course of depression in pregnant women. Conclusions : This study found significant difference in plasma cytokines concentrations between non-pregnant controls and perinatal non-depression controls.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.6
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• pp.460-465
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• 2010

Introduction: A cleft palate is a common birth defect in humans with an incidence of 1/500 to 1/1,000 births. It appears to be caused by multiple genetic and environmental factors during palatogenesis. Many molecules are involved in palate formation but the biological mechanisms underlying the normal palate formation and cleft palate are unclear. Accumulating evidence suggests that transforming growth factor $\beta$/bone morphogenetic proteins (TGF-$\beta$/BMP) family members mediate the epithelial-mesenchymal interactions during palate formation. However, their roles in palatal morphogenesis are not completely understood. Materials and Methods: To understand the roles of TGF-$\beta$/BMP signaling in vivo during palatogenesis, mice with a palatal mesenchyme- specific deletion of Smad4, a key intracellular mediator of TGF-$\beta$/BMP signaling, were generated and analyzed using the Osr2Ires-Cre mice. Results: The mutant mice were alive at the time of birth with open eyelids and complete cleft palate but died within 24 hours after birth. In skeletal preparation, the horizontal processes of the palatine bones in mutants were not formed and resulted in a complete cleft palate. At E13.5, the palatal shelves of the mutants were growing as normally as those of theirwild type littermates. However, the palatal shelves of the mutants were not elevated at E14.5 in contrast to the elevated palatal shelves of the wild type mice. At E15.5, the palatal shelves of the mutants were elevated over the tongue but did not come in contact with each other, resulting in a cleft palate. Conclusion: These results suggest that mesenchymal Smad4 mediated signaling is essential for the growth of palatal processes and suggests that TGF-$\beta$/BMP family members are essential regulators during palate development.

• Women's Health Nursing
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• v.20 no.1
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• pp.14-28
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• 2014

Purpose: This study was to identify relationships of maternal psychosocial factors including mother's mood state, childcare stress, social support and sleep satisfaction with breastfeeding adaptation and immune substances in breast milk, especially secretory immunoglobulin A (sIgA) and transforming growth factor-beta 2 (TGF-${\beta}2$). Methods: Data were collected from 84 mothers who delivered full-term infants by natural childbirth. Structured questionnaires and breast milk were collected at 2~4 days and 6 weeks postpartum. Data were analyzed using descriptive statistics, Pearson's correlation, multiple linear regression, and generalized estimating equation (GEE). Results: Scores for the breastfeeding adaptation scale were significantly related with child care stress, mood state and social support. Mother's anger was positively correlated with the level of sIgA in colostrum (p<.01). Immune substances of breastmilk was significantly influenced by time for milk collection (p<.001) and the type of breastfeeding (sIgA, p<.001, TGF-${\beta}2$, p=.003). Regression analysis showed that breastfeeding adaptation could be explained 59.1% by the type of breastfeeding, childcare stress, the Profile of Mood States, emotional support and sleep quality (F=16.67, p<.001). Conclusion: The findings from this study provide important concepts of breastfeeding adaptation program and explanation of psychosocial factors by immune substances in breast milk. Future research, specially, bio-maker research on breast milk should focus on the ways to improve breastfeeding adaptation.

• Clinical and Experimental Pediatrics
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• v.54 no.6
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• pp.272-275
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• 2011

A recently recognized connective tissue disorder, Loeys-Dietz syndrome (LDS) is a genetic aortic aneurysm syndrome caused by mutations in the transforming growth factor-receptor type I or II gene (TGFBR1 or TGFBR2). They have distinctive phenotypic abnormalities including widely spaced eyes (hypertelorism), bifid uvula or cleft palate, and arterial tortuosity with aortic aneurysm or dissection throughout the arterial tree. LDS is characterized by aggressive and rapid progression of aortic aneurysm. Therefore, the patients with distinct phenotype, marked aortic dilatation and aneurysm at early age should be suspected to be affected by LDS and rapid TGFBR gene analysis should be done. We report one child diagnosed as LDS due to typical phenotypes and two novel missense mutations of the TGFBR2 gene (c.1526G>T and c.1528A>T).

• Molecules and Cells
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• v.38 no.10
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• pp.886-894
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• 2015

Macrophages are divided into two subpopulations: classically activated macrophages (M1) and alternatively activated macrophages (M2). BCG (Bacilli Calmette-$Gu{\acute{e}}rin$) activates disabled $na{\ddot{i}}ve$ macrophages to M1 macrophages, which act as inflammatory, microbicidal and tumoricidal cells through cell-cell contact and/or the release of soluble factors. Various transcription factors and signaling pathways are involved in the regulation of macrophage activation and polarization. We discovered that BCG-activated macrophages (BAM) expressed a new molecule, and we named it Novel Macrophage Activated Associated Protein 1 (NMAAP1). 1 The current study found that the overexpression of NMAAP1 in macrophages results in M1 polarization with increased expression levels of M1 genes, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-${\alpha}$), Interleukin 6 (IL-6), Interleukin 12 (IL-12), Monocyte chemoattractant protein-1 (MCP-1) and Interleukin-1 beta (IL-$1{\beta}$), and decreased expression of some M2 genes, such as Kruppel-like factor 4 (KLF4) and suppressor of cytokine signaling 1 (SOCS1), but not other M2 genes, including arginase-1 (Arg-1), Interleukin (IL-10), transforming growth factor beta (TGF-${\beta}$) and found in inflammatory zone 1 (Fizz1). Moreover, NMAAP1 overexpression in the RAW264.7 cell line increased cytotoxicity against MCA207 tumor cells, which depends on increased inflammatory cytokines rather than cell-cell contact. NMAAP1 also substantially enhanced the phagocytic ability of macrophages, which implies that NMAAP1 promoted macrophage adhesive and clearance activities. Our results indicate that NMAAP1 is an essential molecule that modulates macrophages phenotype and plays an important role in macrophage tumoricidal functions.