Cyanidin-3-glucoside suppresses Th2 cytokines and GATA-3 transcription factor in EL-4 T cells

Allergic disease is dominated by Th2 immune responses. Interleukin (IL)-4 and IL-13, representative Th2 cytokines, play pivotal roles in the pathogenic activation of the Th2 immune response. In this study, we found that cyanidin-3-glucoside chloride (C3G), an anthocyanin suppressed IL-4 and IL-13 produced in activated EL-4 T cells but not Th1 cytokines including IL-2, interferon-gamma, or IL-12. IL-4 and IL-13 mRNA levels and luciferase activation in cells transiently transfected with IL-4 and IL-13 promoter reporter plasmids were significantly inhibited by C3G, suggesting that suppression might be, at least in part, regulated at the transcriptional level. Data from western blot and reverse transcription-polymerase chain reaction analyses of transcription factors involved in cytokine expression suggested that expression of GATA-3, but not T-bet, was downregulated in the nucleus by C3G. Taken together, our data indicate that C3G may has potential as an anti-allergic agent suppressing Th2 activation by downregulating Th2 cytokines and the GATA3 transcription factor in allergies

사람 대장암 세포주인 HCT116과 전립선암 세포주인 PC3의 세포사멸 유도와 항암제 감수성 조절에 있어서의 비타민 C의 영향 및 관련 기전에 관한 연구

비타민C는 잘 알려진 항산화제의 하나로서 세포 대사나 손상 과정 에서 생성된 자유라디칼을 제거하는 역할을 하며, 콜라젠 합성반응 에서 조효소 역할을 하고 있다. 항산화제인 비타민 C는 Chk2-p53-p21Waf1/Cip1 경로를 조절하여 암세포의 세포성장을 억제하는 것으로 알려져 있다. 하지만 비타민C의 암세포에의 항암효과에 대한 많은 연구가 아직까지 수행되고 있고, 그 효능이나 기전에 대해서는 아직 불명확한 상태이다. SIRT1 단백질은 NAD+ 의존적 탈아세틸화 효소로 세포노화 조절 및 수명 연장 및 세포 사멸/생존 등 다양한 생체 내 중요한 역할을 하고 있다. 특히 SIRT1 단백질은 암 억제전사인자인 p53을 탈아세틸화 시킴으로써 p53의 활성을 저해하므로 항암 요법의 표적으로 지목되고 있다. 또한 SIRT1 단백질을 제거한 암세포가 항암제에 민감한 것으로 나타났다. 비타민C가 Chk2-p53-p21Waf1/Cip1 경로를 통해 암세포의 세포성장을 조절하지만, 비타민C가 SIRT1 단백질을 조절하는지에 대해서는 아직 알려지지 않았다. 그러므로 사람 대장암 세포주와 전립선암 세포주에서 비타민C의 SIRT1단백질과 p53단백질에 미치는 영향과 항암제에 대한 민감도에 대한 연구를 수행하였다. 비타민C는 p53유전자의 유무와 무관하게 사람 대장암 세포를 사멸시켰으며, 사람 대장암 세포주와 전립선암 세포주 모두 에서 SIRT1 단백질의 발현을 감소시켰다. 비타민C에 의한 SIRT1 단백질 발현의 감소가 p53 단백질에 영향을 미치지는 않았으나, 비타민 C에 의한 세포사멸에는 p53이 직접적으로 관여하는 것으로 나타났다. 또한 비타민C는 5-FU, 시스플라틴, 독소루비신, 파클리탁셀과 같은 항암제가 암세포의 세포사멸을 유도하는 데 민감도를 높여 주는 것으로 나타났다.Vitamin C is an essential nutrient for living organisms in which it protects the body against oxidative stress. In several tumor cells, it has been reported that vitamin C induces apoptosis and suppresses the proliferation of cancer cells at relatively low concentration (< 1 mM) via the regulation of Chk2-p53-p21Waf1/Cip1 pathway. SIRT1, a NAD-dependent deacetylase, plays an important role in the regulation of energy homeostasis in response to nutrient availability. SIRT1 deacetylates not only histone proteins but also pro-apoptotic proteins such as p53. Overexpression of SIRT1 is implicated in many cancers including colon cancers and SIRT1 silencing induces growth arrest and apoptosis in human cancer cell lines. There are many reports regarding the relationship between SIRT1 and p53 expression and the effect of vitamin C on the regulation of p53 activity, but it still remains to be clarified whether vitamin C regulates SIRT1 expression and its function. Therefore, we have investigated whether vitamin C have an anti-tumor effect through the regulation of SIRT1 in human colon cancer and human prostate cancer. Vitamin C induced apoptosis of HCT116 wt-p53, HCT116 p53-null and PC3, and the expression of SIRT1 mRNA and SIRT1 protein on both HCT116 wt-p53 and HCT116 p53-null were suppressed by vitamin C. In addition, we found that vitamin C suppressed the phosphorylation of both s27 and s47 on SIRT1 of HCT116 wt-p53 and HCT116 p53-null through the suppression of phosphorylation of JNK1. Unexpectedly, in HCT116 wt-p53, p53 which is the most well known substrate of SIRT1 was decreased by vitamin C. However, vitamin C directly, but partially makes contributions of the increase of apoptosis and p53 expression in sirtinol-treated PC3s. To demonstrate that vitamin C influences cancer cell lines via regulation of SIRT1, we transfected HCT116 and PC3 with SIRT1 vector and SIRT1 siRNA and then those cells were treated with vitamin C. SIRT1 overexpression suppressed the reduction of cell proliferation by vitamin C. On the other hand, when cells with SIRT1 siRNA were treated with vitamin C, the proliferation of those cells was inhibited more than vitamin C suppressed. Furthermore, both HCT116 and PC3 treated with vitamin C showed chemosensitivity to 5-FU, cisplatin, doxorubicin and paclitaxel.Docto

Involvement of MAPK signaling cascades in 8-oxo-dG -induced apoptosis of leukemic cell line, KG-1

Thesis(master`s)--서울대학교 대학원 :의학과 약리학전공,2004.Maste

Problem recognition in consumer decision making process of apparel purchase

학위논문(박사)--서울대학교 대학원 :의류학과,1998.Docto

Double intratibial injection of human tonsil-derived mesenchymal stromal cells recovers postmenopausal osteoporotic bone mass

Background and aims: Osteoporosis, which is a disease characterized by weakening of the bone, affects a large portion of the senior population. The current therapeutic options for osteoporosis have side effects, and there is no effective treatment for severe osteoporosis. Thus, we urgently need new treatment strategies, such as topical therapies and/or safe and effective stem cell therapies. Methods: We investigated the therapeutic potential of directly injecting human tonsil-derived mesenchymal stem cells (TMSC) into the right proximal tibias of ovariectomized postmenopausal osteoporosis model mice. Injections were given once (1x) or twice (2x) during the 3-month experimental period. At the end of the experiment, micro -computed tomographic images revealed some improvement in the proximal tibias and more significant improvement in the femoral heads of treated mice. Results: Osteogenic effect was qualitatively and quantitatively more pronounced in TMSC/2x-treated mice. Furthermore, TMSC/2x mice exhibited significant recovery of the serum osteocalcin level, which is pathologically elevated in osteoporosis, and increased serum alkaline phosphatase, which indicates bone formation. TMSC therapy was generally well tolerated and caused no apparent toxicity in the experimental mice. Moreover, TMSC therapy reduced visceral fat. Conclusion: Our results demonstrate that double injection of TMSC directly into the proximal tibia triggers recovery of osteoporosis, and thus could be a potential therapeutic approach for severe bone loss

Optimization of Microenvironments Inducing Differentiation of Tonsil-Derived Mesenchymal Stem Cells into Endothelial Cell-Like Cells

Background: Stem cell engineering is appealing consideration for regenerating damaged endothelial cells (ECs) because stem cells can differentiate into EC-like cells. In this study, we demonstrate that tonsil-derived mesenchymal stem cells (TMSCs) can differentiate into EC-like cells under optimal physiochemical microenvironments. Methods: TMSCs were preconditioned with Dulbecco&amp;amp;apos;s Modified Eagle Medium (DMEM) or EC growth medium (EGM) for 4 days and then replating them on Matrigel to observe the formation of a capillary-like network under light microscope. Microarray, quantitative real time polymerase chain reaction, Western blotting and immunofluorescence analyses were used to evaluate the expression of gene and protein of EC-related markers. Results: Preconditioning TMSCs in EGM for 4 days and then replating them on Matrigel induced the formation of a capillary-like network in 3 h, but TMSCs preconditioned with DMEM did not form such a network. Genome analyses confirmed that EGM preconditioning significantly affected the expression of genes related to angiogenesis, blood vessel morphogenesis and development, and vascular development. Western blot analyses revealed that EGM preconditioning with gelatin coating induced the expression of endothelial nitric oxide synthase (eNOS), a mature EC-specific marker, as well as phosphorylated Akt at serine 473, a signaling molecule related to eNOS activation. Gelatin-coating during EGM preconditioning further enhanced the stability of the capillary-like network, and also resulted in the network more closely resembled to those observed in human umbilical vein endothelial cells. Conclusion: This study suggests that under specific conditions, i.e., EGM preconditioning with gelatin coating for 4 days followed by Matrigel, TMSCs could be a source of generating endothelial cells for treating vascular dysfunction

Expression of tenocyte lineage-related factors from tonsil-derived mesenchymal stem cells

Human palatine tonsil-derived mesenchymal stem cells (TMSCs) are known to be a new source of progenitor cells. Using waste tissue after tonsillectomy as a cell provider can be the biggest benefit of TMSCs, compared with other stem cells. The purpose of this study was to investigate tenogenic differentiation of TMSCs and to access the differential effects of transforming growth factor beta 3 (TGF-β3) on the tenogenesis of TMSCs. Human tonsil was obtained after tonsillectomy. Using a cytometric analysis, we were able to find that the TMSCs had typical mesenchymal stem cell markers: positive for CD73, CD90, and CD105, and negative for CD14, CD34, and CD45. Using TGF-β3, the expressions of tenocyte-specific genes and proteins, such as collagen type 1 (COL1), tenomodulin (TNMD), and scleraxis (SCX), were measured by a quantitative polymerase chain reaction (PCR), immunofluorescence staining, immunohistochemistry and Western blot analyses. Quantitative PCR assay showed that TGF-β3 significantly increased the expressions of tenocyte lineage marker genes, including COL1, TNMD, and SCX, at a 3-day treatment, compared with control. However, these increases were not found at long-term exposures (7 or 10 days), except that TNMD expression was maintained at 50 ng/mL at a 7-day exposure to TGF-β3. Like genes, the protein expression levels of COL1, TNMD, and SCX were also induced in TGF-β3-treated TMSCs in a 3-day treatment, which were maintained for 10 days, as evidenced by immunofluorescence staining, immunohistochemistry and Western blot analyses. This study demonstrated that TMSCs in tenogenic stimulation with TGF-β3 have a high tenogenic differentiation potential. © 2016, The Korean Tissue Engineering and Regenerative Medicine Society and Springer Science+Business Media Dordrecht

Autophagy induction in the skeletal myogenic differentiation of human tonsil-derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation and are thus a valuable source for the replacement of diseased or damaged organs. Previously, we reported that the tonsils can be an excellent reservoir of MSCs for the regeneration of skeletal muscle (SKM) damage. However, the mechanisms involved in the differentiation from tonsil-derived MSCs (T-MSCs) to myocytes via myoblasts remain unclear. To clarify these mechanisms, we analyzed gene expression profiles of T-MSCs during differentiation into myocytes compared with human skeletal muscle cells (hSKMCs). Total RNA was extracted from T-MSCs, T-MSC-derived myoblasts and myocytes, and hSKMCs and was subjected to analysis using a microarray. Microarray analysis of the three phases of myogenic differentiation identified candidate genes associated with myogenic differentiation. The expression pattern of undifferentiated T-MSCs was distinguishable from the myogenic differentiated T-MSCs and hSKMCs. In particular, we selected FNBP1L, which among the upregulated genes is essential for antibacterial autophagy, since autophagy is related to SKM metabolism and myogenesis. T-MSCs differentiated toward myoblasts and skeletal myocytes sequentially, as evidenced by increased expression of autophagy-related markers (including Beclin-1, LC3B and Atg5) and decreased expression of Bcl-2. Furthermore, we reconfirmed that autophagy has an effect on the mechanism of skeletal myogenic differentiation derived from T-MSCs by treatment with 5-azacytidine and bafilomycin A1. These data suggest that the transcriptome of the T-MSC-derived myocytes is similar to that of hSKMCs, and that autophagy has an important role in the mechanism of myogenic differentiation of T-MSCs

3D Culture of Tonsil-Derived Mesenchymal Stem Cells in Poly(ethylene glycol)-Poly(L-alanine-co-L-phenyl alanine) Thermogel

Poly(ethylene glycol)-poly(L-alanine-co-L-phenyl alanine) (PEG-PAF) aqueous solutions undergo sol-to-gel transition as the temperature increases. The transition is driven by the micelle aggregation involving the partial dehydration of the PEG block and the partial increase in beta-sheet content of the PAF block. Tonsil-tissue-derived mesenchymal stem cells (TMSCs), a new stem cell resource, are encapsulated through the sol-to-gel transition of the TMSC-suspended PEG-PAF aqueous solutions. The encapsulated TMSCs are in vitro 3D cultured by using induction media supplemented with adipogenic, osteogenic, or chondrogenic factors, where the TMSCs preferentially undergo chondrogenesis with high expressions of type II collagen and sulfated glycosaminoglycan. As a feasibility study of the PEG-PAF thermogel for injectable tissue engineering, the TMSCs encapsulated in hydrogels are implanted in the subcutaneous layer of mice by injecting the TMSC suspended PEG-PAF aqueous solution. The in vivo studies also prove that TMSCs undergo chondrogenesis with high expression of the chondrogenic biomarkers. This study suggests that the TMSCs can be an excellent resource of MSCs, and the thermogelling PEG-PAF is a promising injectable tissue engineering scaffold, particularly for chondrogenic differentiation of the stem cells