• Journal of Nutrition and Health
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• 제49권5호
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• pp.277-287
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• 2016

본 연구는 제5기 2차년도 국민건강영양조사 결과를 활용하여 한식 식품군의 총 페놀 함량, in vitro 항산화활성 및 인체세포를 이용한 ex vivo DNA 손상 감소효과를 비교하고, 각 지표간의 상관성을 분석하며, 한식의 총 식사 항산화능에 대한 각 식품군의 기여도를 알아보기 위해 수행되었다. 제5기 2차년도 국민건강영양조사 결과를 바탕으로 한식의 식물성 식품을 10가지 식품군 (곡류, 과일류, 채소류, 견과류, 김치류, 해조류, 감자류, 버섯류, 두류, 오일류)으로 분류한 후 각 식품군별로 총 섭취량의 1% 이상 섭취한 식품 84종을 한식의 식물성 식품으로 최종 선정하였다. 각 식품군의 총 페놀함량을 측정하였고, DPPH radical scavenging assay, TEAC assay, $ORAC_{ROO{\cdot}}$ assay를 사용하여 in vitro 항산화능을 측정하였다. 한식의 식품군별 항산화능 (dietary antioxidant capacity, DAC)은 in vitro 항산화활성 평균값과 각 식품군의 1일 섭취량을 고려하여 계산하였고 한식 TDAC는 각 식품군의 DAC로의 합으로 구하였으며, TDAC에 대한 각 식품군 항산화능의 기여도를 평가하였다. 인체 임파구에서의 ex vivo DNA 손상 정도는 comet assay를 사용하여 평가하였다. 한식 식품군의 총 페놀함량은 버섯류, 과일류, 채소류, 해조류, 김치류 등의 순으로 높았으며, 3가지 in vitro 실험법을 평균한 식품군의 항산화활성 순위는 버섯류, 해조류, 채소류, 김치류, 과일류 등의 순이었다. 각 식품군의 항산화활성에 식품섭취량을 고려하여 계산한 한식의 TDAC에 대한 식품군의 항산화능 기여도는 곡류가 33.4%로 가장 높았으며, 과일류 (23.9%), 채소류 (12.7%), 김치류 (11.2%) 등의 순으로 나타났다. 인체 임파구에서 ex vivo DNA 손상 보호효과는 버섯류에서 가장 높았으며, 그 다음 채소류, 과일류, 해조류, 김치류의 순으로 나타났다. 각 식품군의 페놀함량과 in vitro 항산화 활성, 그리고 ex vivo DNA 보호효과의 순위가 비슷하게 나타났으며 각 지표간의 상관성은 매우 높았다. 한식 식품군 중 버섯류, 과일류, 채소류, 해조류에서 총 페놀함량과 항산화 활성, DNA 손상 보호효과가 높게 나타났다. 각 식품군의 총 페놀함량과 in vitro 항산화 활성, ex vivo DNA 보호효과 지표 간의 상관성은 매우 높았다. 한식의 TDAC에 대한 식품군별 항산화능 기여도는 곡류가 가장 높았고, 그 다음이 과일류, 채소류, 김치류의 순이었다. 이러한 결과는 앞으로 한식의 우수성을 항산화 측면에서 밝히는데 매우 중요한 기초자료로 활용될 수 있을 것이다.

• Journal of Periodontal and Implant Science
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• 제44권4호
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• pp.158-168
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• 2014

Purpose: The present study was conducted to evaluate the impact of scaling and root planing (SRP) on the C-reactive protein (CRP) levels of gingival crevicular fluid (GCF) and serum in chronic periodontitis patients with type 2 diabetes mellitus (T2DM-CP) or without type 2 diabetes mellitus (NDM-CP). Methods: Forty-eight human participants were divided into two groups: an experimental (T2DM-CP) group (group I, n=24) comprising chronic periodontitis patients with random blood sugar ${\geq}200mg/dL$ and type 2 diabetes mellitus, and control (NDM-CP) group (group II, n=24) of those with chronic periodontitis and random blood sugar <200 without T2DM for the study. All subjects underwent nonsurgical periodontal therapy (NSPT) including complete SRP and subgingival debridement. Periodontal health parameters, plaque index (PI), gingival index (GI), probing pocket depth (PPD), clinical attachment level (CAL), GCF volume (GCF vol), GCF-CRP, random blood glucose (RBS), glycated hemoglobin, and systemic inflammatory markers, serum CRP, total leukocyte count (TLC), neutrophil count (Neutr) and lymphocyte count (Lymph), were evaluated at baseline, 1 month, and 3 months after SRP. Results: NSPT resulted in statistically significant improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum as well as GCF of both groups I and II. The mean improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum and GCF was greater in group I than group II after NSPT. There was nonsignificant increase in GCF-CRP, TLC, Lymph, and RBS, and a significant increase in Neutr and Serum CRP in group II at 1 month. The Serum CRP level of 20 out of 24 group II patients had also increased at 1 month. Conclusions: The CRP levels in both GCF and serum were higher in T2DM-CP patients than in NDM-CP patients. Although there was a significant improvement in both the groups, greater improvement was observed in both GCF and serum samples of T2DM-CP patients.

• 한국식품영양학회지
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• 제28권1호
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• pp.34-39
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• 2015

본 연구는 식용 버섯의 조리방법에 따른 항산화 생리활성의 평가를 위해 수행되었으며, 산화적 스트레스에 의한 DNA 손상 감소 효과를 통해 조리방법을 달리한 버섯 추출물의 유전독성학적 방호효과를 살펴보았다. Human lymphocyte에 조리방법을 달리한 3가지 버섯(느타리, 팽이, 표고)의 추출물을 처리하고, hydrogen peroxide($H_2O_2$)로 산화적 손상을 준 후, DNA 감소 효과를 Comet assay로 평가한 결과, 모든 시료군에서 산화적 손상에 의한 DNA 손상 감소 효과를 나타냈다. 3가지 버섯 모두 비조리군이 조리군보다 높은 효과를 나타냈는데, 이는 조리과정에 의한 페놀성 화합물의 감소로 인한 것으로 보이며, 조리군 중에서 볶기와 전이 비교적 낮은 DNA 손상 감소 효과를 나타낸 것은 조리 시 첨가되었던 대두유의 가열 산화에 의한 것으로 사료된다. 결론적으로, 조리된 버섯은 생버섯에 비해 산화적 스트레스에 의한 DNA 손상 감소효과가 낮으나, 양성 대조군과 비교하였을 때 손상을 유의적으로 감소시킨 것으로 나타났다. 또한, 본 연구에서 사용한 네 가지 조리법(굽기, 데치기, 볶기, 전) 중 DNA 손상 감소에 효과적인 조리법은 대두유를 사용하지 않은 굽기와 데치기인 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.265-271
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• 2014

Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive $CD8^+$ T cells with induction of IFN-${\gamma}$ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-${\gamma}$ levels were measured by ELISA and flow cytometry, respectively. $CD8^+$ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-${\gamma}$ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of $CD8^+$ T cells from tumor bearing mice, while anti-IFN-${\gamma}$ blocked the function of $CD8^+$ T cells, suggesting that IFN-${\gamma}$ mediated the cytolytic activity of $CD8^+$ T cells. Furthermore, in vivo neutralization of $CD8^+$ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating $CD8^+$ T cells and stimulating them to secrete IFN-${\gamma}$ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.

• KSBB Journal
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• 제20권4호
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• pp.278-284
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• 2005

식물세포의 느린 생장과 낮은 생산량의 이유로 지금까지 는 주로 미생물이나 동물세포에서 유전자 재조합 단백질을 생산하여 왔다. 그러나 저렴한 배지 가격, 동물 유래 바이러스 감염 위험성으로부터의 안정성, glycosylation 등의 post-translational modification이 가능하다는 장점들로 인하여 최근 들어 식물세포배양은 생물학적 활성을 가진 고부가가치의 단백질을 생산하는데 많이 이용되고 있다. 본 연구에서는 생장배지에 첨가했던 sucrose의 소비와 induction 배지로의 교환에서 오는 배지내의 삼투압을 조절하여 hCTLA4-Ig의 생산성을 높이고자 하였다. 다양한 삼투압 조절제 첨가 실험을 통해 sorbitol을 선별하고, 40 mM의 sorbitol 첨가에서 상대적으로 높은 생존도와 induction 후 7일째 대조구보다 1.7배 높은 생산성을 확인하였다. 또한, 저농도의 glucose 첨가를 통한 생산성 증대에 있어서는 8 mM glucose에서 induction 이후에도 높은 세포농도를 유지하면서 최대 37.3 mg/L까지 hCTLA4-Ig 생산량을 증가시켰다. 5-L bioreactor에서 회분식 배양과 induction시의 hCTLA4-Ig 생산량을 비교한 결과 induction시 배양 18일째 최고 45.3 mg/L까지 높일 수 있었으며, 회분식 배양에 비해 2.1배 증가됨을 확인하였다.

• KSBB Journal
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• 제31권4호
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• pp.277-283
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• 2016

In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

• 한국환경성돌연변이발암원학회지
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• 제22권2호
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• pp.112-124
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• 2002

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to exert detrimental toxicities on various organ systems including reproductive, cardiovascular, nervous, or dermal system. Immunomodulatory effects of TCDD is thymic atrophy, downregulation of cytotoxic T or B lymphocyte differentiation and activation, which were demonstrated using experimental animals, whereas immunotoxicity in human has not been investigated well. This study was proceeded to evaluate general immunologic spectrum of the Korean Vietnam War veterans exposed to TCDD during their operation, and compare with that of the non-exposed control subjects with similar age. Regarding composition and quantity, immune cells in peripheral blood collected from the TCDD-exposed was not much different from those of the control except decreased red blood cell, hemoglobin and hematocrit level. Furthermore, plasma IgG2, G3, and G4 isotype distribution was similar between two groups, but IgG1 level was significantly lowered in the TCDD-exposed, indicating a TCDD-mediated functional alteration of B cells. Significantly enhanced level of IgE in plasma, a hallmark of dermal or respiratory allergic response, was also observed in the TCDD-exposed compared with that of the control. Elevated generation of IL-4 and IL-10 was resulted from in vitro stimulation of T cells with PMA plus ionomycin or PHA, respectively, from the TCDD-exposed in comparison to those of the control, suggesting a skewed type-2 response. In addition, the level of IFN${\gamma}$, a multifunctional cytokine for T cell-mediated immunity, was lowered in the TCDD-exposed with upregulation of tumor necrosis factor $\alpha$. The present study suggests that TCDD exposure disturbs immunohomeostasis in humans observed as an aberrant plasma IgE and IgG1 levels and dysregulation of T cell activities.

• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.116-123
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• 2005

The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

• Journal of Nutrition and Health
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• 제36권2호
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• pp.125-132
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• 2003

The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제26권1호
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• pp.5-17
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• 2000

To evaluate the possible therapeutic effects of growth hormone and vitamin C on multiorgan failure, a rat model was developed for LPS-induced sepsis. Using this model, the effects of growth hormone and vitamin C on tissue damages, catalase and i-NOS activities, and MDA levels were examined in the lung and liver. The level of TNF- in plasm was also examined. Male, Sprague-Dawley rats were injected with LPS intraperitoneally then divided into 3 groups : positive controls injected with LPS only, the ones injected with growth hormone or vitamin C immediately after the LPS injections. The lung and the liver were then isolated, blood samples were collected at 24 or 48 hours after the LPS injection, then examined for histopathological and biochemical changes. The results obtained were as follows. 1. LPS induced sinusoid vasodilation and mild destruction of lobular structure in the liver. In the lung, alveolar structure appeared to be thickened and interstitial edema was observed. The levels of MDA in the liver and the lung was increased by LPS, while the activity of catalase was decreased. The activity of i-NOS of those tissues was also increased, which was more pronounced at 24 hr. The level of TNF- in plasm was increased by LPS 2. In the lung, vitamin C suppressed lymphocyte and neutrophil infiltration, alveolar wall thickening and interstitial edema. In the liver, vitamin C protected against the destruction of the lobular structure. The activity of catalase reduced by LPS was reversed partly by vitamin C. The activity of i-NOS enhanced by LPS was also reversed by vitamin C. The level of TNF- in plasm reduced in some animals by vitamin C, which however was not significant statistically(p<0.05). 3. Growth hormone showed similar protective effects against inflammation and damages in the liver and lung tissues. Growth hormone reversed partly the LPS effects on the level of MDA, the activity of catalase and i-NOS induction in the liver and the lung. Growth hormone reduced plasma level of TNF-${\alpha}$ substantially, which contrasted from vitamin C. Besides this, overall protective effects of growth hormone against LPS-induced experimental sepsis were similar to those of vitamin C. From this results, the mechanism of growth hormone on suppression of LPS-induced tissue damage might be associated with production of antioxidative enzyme and suppression of plasma TNF- level.