• Food Science and Biotechnology
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• 제18권6호
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• pp.1481-1486
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• 2009

The immunomodulating effect of Salicornia herbacea polysaccharides on BALB/c mice splenocytes was investigated. Crude (CS) and fine polysaccharide (PS) extracts with potential biological activity were prepared from S. herbacea. For in vitro experiments, splenocytes and separated T cells were treated with CS and PS (0.5, 1, 2, and 4 mg/mL). For in vivo experiments, the CS and PS were orally administered to BALB/c mice every day for 2 weeks. For basic data analysis, physiological parameters were recorded. Cell proliferation of splenocytes and T cells was used as an index for immunomodulating activity. The proliferation of splenocytes and separated T cells was 3.2 and 3.5 times higher than the control, respectively. Moreover, when splenocytes were treated with mitogen, the highest proliferation rate was observed in splenocytes cultured with PS. Interestingly, the stimulative activity of PS was more strongly exerted through $CD4^+$ T cells than through $CD8^+$ T cells.

• Natural Product Sciences
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• 제17권3호
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• pp.212-215
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• 2011

In the course of screening anti-adipogenic activity of natural products employing the preadipocyte cell line, 3T3-L1 as an in vitro assay system, the EtOAc fraction of the stem barks of Acer tegmentosum Maxim (Aceraceae) showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Activity-guided fractionation led to the isolation of active constituent, (+)-catechin. (+)-Catechin showed inhibitory activity on adipocyte differentiation in dose-dependent manner. Further studies with interval treatment demonstrated that (+)-catechin exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Our present study also showed that (+)-catechin significantly inhibited the preadipocyte proliferation. Taken together, these results suggest that (+)-catechin, a constituent of A. tegmentosum might contribute the anti-adipogenic activity of A. tegmentosum.

• Journal of Animal Science and Technology
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• 제62권6호
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• pp.854-863
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• 2020

This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

• Tuberculosis and Respiratory Diseases
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• 제86권4호
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• pp.304-318
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• 2023

Background: Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and significantly contribute to immune evasion. We investigated the effects of CAFs on the immune function of CD4+ and CD8+ T cells in non-small cell lung cancer (NSCLC). Methods: We isolated CAFs and normal fibroblasts (NFs) from tumors and normal lung tissues of NSCLC patients, respectively. CAFs were co-cultured with activated T cells to evaluate their immune regulatory function. We investigated the effect of CAF conditioned medium (CAF-CM) on the cytotoxicity of T cells. CAFs were also co-cultured with activated peripheral blood mononuclear cells and further incubated with cyclooxygenase-2 (COX2) inhibitors to investigate the potential role of COX2 in immune evasion. Results: CAFs and NFs were isolated from the lung tissues (n=8) and lymph nodes (n=3) of NSCLC patients. Immune suppressive markers, such as COX2 and programmed death-ligand 1 (PD-L1), were increased in CAFs after co-culture with activated T cells. Interestingly, CAFs promoted the expression of programmed death-1 in CD4+ and CD8+ T cells, and strongly inhibited T cell proliferation in allogenic and autologous pairs of CAFs and T cells. CAF-CM decreased the cytotoxicity of T cells. COX2 inhibitors partially restored the proliferation of CD4+ and CD8+ T cells, and downregulated the expression of COX2, prostaglandin E synthase, prostaglandin E2, and PD-L1 in CAFs. Conclusion: CAFs promote immune evasion by suppressing the function of CD4+ and CD8+ T cells via their effects on COX2 and PD-L1 in NSCLC. The immunosuppressive function of CAFs could be alleviated by COX2 inhibitors.

• Journal of Nutrition and Health
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• 제55권1호
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• pp.47-58
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• 2022

Purpose: Obesity and a high-fat diet (HFD) are risk factors for colorectal cancer. We have previously shown that luteolin (LUT) supplementation in HFD-fed mice markedly inhibits tumor development in chemically induced colon carcinogenesis. In this study, we evaluated the anticancer effect of LUT in the inhibition of cell proliferation in HFD-fed obese mice and HT-29 human colorectal adenocarcinoma cells grown in an adipocyte-derived medium. Methods: C57BL/6 mice were fed a normal diet (ND, 11.69% fat out of total calories consumed, n = 10), HFD (40% fat out of total calories consumed, n = 10), HFD with 0.0025% LUT (n = 10), and HFD with 0.005% LUT (n = 10) and were subjected to azoxymethane-dextran sulfate sodium chemical colon carcinogenesis. All mice were fed the experimental diet for 11 weeks. 3T3-L1 preadipocytes and HT-29 cells were treated with various doses of LUT in an adipocyte-conditioned medium (Ad-CM). Results: The weekly body weight changes in the LUT groups were similar to those in the HFD group; however, the survival rates of the LUT group were higher than those of the HFD group. Impaired crypt integrity of the colonic mucosa in the HFD group was observed to be restored in the LUT group. The colonic expression of proliferating cell nuclear antigen and insulin-like growth factor 1 (IGF-1) receptors were suppressed by the LUT supplementation in the HFD-fed mice. The LUT treatment (10, 20, and 40 µM) inhibited the proliferation and migration of HT-29 cells cultured in Ad-CM in a dose-dependent manner, as well as the differentiation of 3T3-L1 preadipocytes. Conclusion: These results suggest that the anticancer effect of LUT is probably due to the inhibition of IGF-1 signaling and adipogenesis-related cell proliferation in colon cancer cells.

• 한국식품과학회지
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• 제39권6호
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• pp.701-707
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• 2007

민간요법에서 항당뇨 및 항비만 효과가 있는 것으로 알려진 길경의 항당뇨 효과가 있는 지 여부를 in vitro에서 조사하기 위해서 길경을 70% 에탄올로 추출한 후 메탄올과 물을 섞은 용액으로 단계별로 XAD-4 column으로 분획하였다. 본 연구에서는 1) 3T3-L1 섬유아세포와 지방세포에서 길경의 추출 분획물이 인슐린처럼 작용하는 인슐린성 물질이거나, 2) 인슐린 작용을 향상시키는 인슐린 민감성 물질이거나, 또는 3) 포도당 자극에 의한 인슐린 분비를 향상시키거나, 4) 베타세포의 기능과 양을 증가시키는데 관여하는 유전자인 IRS-2, glucokinase, PDX-1의 mRNA 발현을 향상시키거나, 5) $\alpha-glucoamylase$ 활성을 억제하는 물질로 작용하는 지 여부를 조사하였다. 길경 추출 분획물은 인슐린성 물질로 작용하지 않았다. 반면에 0, 20와 100%메탄올층은 3T3-L1 지방세포에서 인슐린 자극에 의한 포도당 흡수를 증가시켰다. 이 분획층 중에서 특히 0%과 100% 메탄올 분획층은 분화 유도물질의 작용을 향상시켜 3T3-L1 섬유아세포에서 지방세포로의 분화 및 중성 지방의 축적을 증가시켰다. 그러므로 이들은 $PPAR-{\gamma}$ agonist로 작용하는 물질을 함유할 가능성이 매우 높다. 인슐린을 분비하는 세포인 Min6 세포에서 포도당 자극에 의한 인슐린 분비를 향상시키는 지 여부를 조사하였는데 20, 80 그리고 100% 메탄올층은 포도당 자극에 의한 인슐린 분비를 증가시켰다. 그 기전은 인슐린 분비와 베타세포의 증식에 관여하는 유전자의 IRS-2, glucokinase 그리고 PDX-1의 mRNA의 양을 증가시키는 것과 관련이 있다. 결론적으로 길경은 지방 세포의 분화를 촉진하는 물질, 인슐린 민감성을 향상시키는 물질 그리고 베타세포의 기능과 증식을 촉진시키는 물질을 함유하고 있으므로 우리나라 및 아시아의 사람들에서 많이 유발되는 비만을 동반하지 않은 당뇨병 및 인슐린 저항성의 치료와 예방에 중요한 역할을 할 것으로 사료된다.

• Natural Product Sciences
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• 제16권4호
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• pp.291-294
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• 2010

In obesity, adipocytes undergo abnormal growth characterized by increased cell numbers and differentiation. Thus, inhibition of mitogenesis of preadipocytes and their differentiation to adipocytes would be beneficial for the prevention and progression of obesity. In the present study, we attempted to evaluate antiadipogenic activity of adlay (Coix seed, the seed of Coix lacryma-jobi L. var. ma-yuen Stapf) employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. Because several varieties of adlay are in use in Korea, anti-adipogenic activity of three varieties of adlay such as Sang-Gang, Jo-Hyun and Yulmu-Ilho was evaluated. All the three varieties of adlay showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Adlay, however, showed little effects on adipocyte proliferation. Further studies with interval treatment demonstrated that adlay exerted inhibitory activity on adipocyte differentiation via acting on early stage of adipogenesis. Taken together, adlay might be useful in the prevention of obesity.

• 한국식품영양과학회지
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• 제45권8호
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• pp.1107-1113
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• 2016

지방전구세포가 지방세포로 분화하는 과정에서 아선약 추출물의 영향을 확인하고자 분화유도제를 처리하여 3T3-L1 세포에서 아선약의 세포독성, 지방축적 억제 효과, triglyceride 억제 효능, 지방분화 관련 단백질들의 발현을 확인하였다. 아선약 추출물을 최대 $200{\mu}g/mL$ 농도까지 세포에 처리하여 세포독성을 측정한 결과 아선약 추출물은 $100{\mu}g/mL$ 이하의 농도에서는 세포에 독성을 유발하지 않는 것으로 확인되었다. Oil red-O 염색을 통해 지방축적률을 확인한 결과 아선약 추출물에 의한 농도 의존적인 지방축적의 감소 효과가 있었다. Triglyceride 생성량 역시 농도 의존적인 감소가 확인되었다. 아선약에 의한 지방분화의 감소가 어떠한 기작에 의해 조절되는지 확인하고자 관련 지표단백질인 $PPAR{\gamma}$와 SREBF-1 그리고 $C/EBP{\alpha}$의 발현량 변화를 확인해본 결과 $100{\mu}g/mL$ 농도에서 유의성 있게 감소하였다. 이상의 결과를 종합해볼 때 아선약 추출물은 지방세포의 분화억제와 분화된 지방세포의 지방축적을 저해함으로써 항비만 효과를 유도할 것으로 기대된다.

• 대한한의학회지
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• 제24권3호
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• pp.35-44
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• 2003

Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.

• Nutrition Research and Practice
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• 제15권4호
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• pp.444-455
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• 2021

BACKGROUND/OBJECTIVES: Adipocytes undergo angiogenesis to receive nutrients and oxygen needed for adipocyte' growth and differentiation. No study relating quercetin with angiogenesis in adipocytes exists. Therefore, this study investigated the role of quercetin on adipogenesis in 3T3-L1 cells, acting through matrix metalloproteinases (MMPs). MATERIALS/METHODS: After proliferating preadipocytes into adipocytes, various quercetin concentrations were added to adipocytes, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to evaluate cell proliferation. Glycerol-3-phosphate dehydrogenase (GPDH) activity was investigated as an indicator of fat accumulation. The mRNA expressions of transcription factors related to adipocyte differentiation, CCAAT/enhancer-binding proteins (C/EBPs), peroxisomal proliferatoractivated receptors (PPAR)-γ, and adipocyte protein 2 (aP2), were investigated. The mRNA expressions of proteins related to angiogenesis, vascular endothelial growth factor (VEGF)-α, vascular endothelial growth factor receptor (VEGFR)-2, MMP-2, and MMP-9, were investigated. Enzyme activities and concentrations of MMP-2 and MMP-9 were also measured. RESULTS: Quercetin treatment suppressed fat accumulation and the expressions of adipocyte differentiation-related genes (C/EBPα, C/EBPβ, PPAR-γ, and aP2) in a concentration-dependent manner in 3T3-L1 cells. Quercetin treatments reduced the mRNA expressions of VEGF-α, VEGFR-2, MMP-2, and MMP-9 in 3T3-L1 cells. The activities and concentrations of MMP-2 and MMP-9 were also decreased significantly as the concentration of quercetin increased. CONCLUSIONS: The results confirm that quercetin inhibits adipose tissue differentiation and fat accumulation in 3T3-L1 cells, which could occur through inhibition of the angiogenesis process related to MMPs.