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홍삼추출물이 마우스 복강 대식세포 Hydrogen Peroxide 생산에 미치는 영향
박란숙 한국식품영양학회 1998 韓國食品營養學會誌 Vol.11 No.1
홍삼의 추출물인 50% ethanol extract, crude saponin, 그리고 lipid soluble fraction이 마우스 대식세포의 oxidative burst를 유발할 수 있는지 여부를 알아보고자 in vitro와 in vivo에 각각의 추출물을 처치하고 hydrogen peroxide 생산을 DCFH-DA를 이용한 형광분광광도법으로 측정하였다. 형광분광법에 의한 hydrogen peroxide의 측정을 최적화하기 위한 DCFH-DA의 농도는 3.2 μM이었고, oxidative burst를 유발하지 못하였지만, zymosan A로 유발한 경우에는 50% ethanol extract에서 가장 높은 hydrogen peroxide를 생산하였다. In vivo 실험에서는, lipid soluble extract에서만 유의하게 증가한(P<0.01) oxidative burst를 유발하였고, ginsenoside(saponin)가 어느 정도 포함되어 있는 50% ethanol extract와 crude saponin은 대조군에 비하여 유의하게 낮은(P<0.05) hydrogen peroxide를 생산하였다. 이는 ginsenoside가 마우스의 nitric oxide 생산을 억제한다는 다른 연구자들의 보고와 일치하는 결과이다. Oxidative burst를 유발한 lipid soluble extract에는 phenol계 화합물, polyacetylene계 화합물, 미량선분 등이 함유되어 있으므로 차후 연구를 통하여 과연 어느 성분이 hydrogen peroxide를 증가시키는지를 규명하는 것이 필요하다. This experiment has conducted to evaluate whether single injection of red ginseng extract including 50% ethanol extract, crude saponin, and lipid soluble fraction can induce oxidative burst of mouse peritoneal macrophages with use of fluorescence spectrophotometer. To optimize conditions of fluorescent spectrophotometry concentrations of DCFH-DA(2',7'-dichlorofluorescin diacetate) was 1.6 μM and control oxidative burst by Zymosan A and PMA(phorbol myristate acetate) were 100㎍, 250ng, respectively. Though in vitro macrophages failed to induce increment of H_2O_2 production, but 50% ethaol extract group induced significant enhancement of H_2O_2 production when zymosan A triggered oxidative burst. On the other hand, lipid soluble fraction enhanced significantly H_2O_2 production with single peritoneal injection in vivo. Both of 50% ethanol extract and crude saponin group decreased H_2O_2 production than that of control group. These findings consisted with the other reports which showed ginsenosides inhibited nitric oxide production and lipid soluble fraction activated colony stimulating factor(granulocyte-monocyte) activity in bone marrow stem cells. As is well known, lipid soluble fraction contains phenol compound, polyacetylene compound and alkaloids. Further study would unravel which component of it can induce H_2O_2 production of macrophages.
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디엠프리(녹차추출물)에 의한 나균 감염 중간엽줄기세포의 IL-6 생산 억제
박란숙,Park, Ran-Sook 한국식품영양학회 2015 韓國食品營養學會誌 Vol.28 No.4
Previous reports revealed that DMfree (green tea extract) inhibited expression of the IL-6 gene in Mycobacterium lepraeinfected MSCs (mesenchymal stem cells). This study aimed to measure IL-6, $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2 production in M. leprae-infected MSCs using ELISA. To confirm the effect of DMfree on IL-6 and signal transduction, a western blotting test was performed. DMfree inhibited the expression of IL-6 in the MSCs and the heterodimer of STAT3, which also affects the expression of multiple genes. Though DMfree pre-treatment of control MSCs produced a baseline level of IL-6, it significantly inhibited the production of IL-6 in M. leprae-infected MSCs. There was no significant difference in IL-6 production between 1 and 7 day treatment groups. M. leprae-infected MSCs produced more $IL-1{\beta}$, $TNF-{\alpha}$ and PGE2, but DMfree could not inhibit their production at a physiological concentration. This is different from other reports that used higher concentration of EGCG treatment, resulting in significant inhibition of the cytokines. The inhibition appears to be related to the concentration of EGCG. These results indicate that DMfree can alleviate inflammation involving IL-6.
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마늘성분 SAC 및 Hydrogen Peroxide에 의한 줄기세포의 유전자 발현 윤곽
박란숙,Park, Ran-Sook 한국식품영양학회 2012 韓國食品營養學會誌 Vol.25 No.4
Though hydrogen peroxide ($H_2O_2$) causes a deleterious effect to cells with its reactive oxygen species resulting in cell death, S-allyl cysteine (SAC, a bioactive organosulfur compound of aged garlic extract) has been known to have a cytoprotective effect. Few reported profiles of gene expression of $H_2O_2$ and SAC treated human cord blood derived mesenchymal stem cells (MSC). This study revealed changes in the profile of twenty-one genes grouped by oxidative stress, antioxidant, cell death, anti-apoptosis and anti-aging by quantitative real time PCR. A concentration of $100{\mu}M$ of SAC or $50{\mu}M$ of $H_2O_2$ was applied to MSC which show moderate growth and apoptosis pattern. $H_2O_2$ treatment enhanced expression of eleven genes out of twenty-one genes compared with that of control group, on the contrary SAC suppressed expression of eighteen genes out of twenty-one genes except C ros oncogene. SAC decreased expression of oxidative stress genes such as SOD1, CAT and GPX. These results seemed consistent with reports which elucidated over-expression of NF-${\kappa}$B by $H_2O_2$, and suppression of it by SAC. This study will confer basic information for further experiments regarding the effects of SAC on gene levels.
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마늘의 Allicin이 사람 단핵세포의 사이토카인 생산 유전자의 발현에 미치는 영향
박란숙 한국식품영양학회 2002 韓國食品營養學會誌 Vol.15 No.3
마늘의 주요 성분인 allicin 투여 후 유도되는 사람 말초혈액의 단핵구의 유전자 발현에 미치는 allicin 의 효과를 규명하였다. DNA microarray를 이용하여, allicin이 chemokines, cytokine 면역관련 유전자 및 신호전달 관련 유전자의발현을 유도하는 것을 확인하였다. 반대로 allicin은 Thl type의 획득면역 관련 유전자의 발현을 억제하였다. 염증세포에 있어서 allicin은 억제효과 및 자극 효과를 동시에 보여주었다. 이는 allicin이 휴지기 세포에서 먼저 증가시킨 특정 유전자의 발현을 이후에 감소시키는 결과를 보여주는 것으로 positive와 negative 효과를 발휘하는 새로운 기전을 제시하는 것이다. Allicin에 대한 광범위하고 새로운 관심을 고려해 볼 때 본 연구에서 보여주는 많은 유전자의 발현 양상은 좀 더 특정적이고 효과적인 치료법을 고안하는 데 유용할 것이다. The effect of allicin, the major component of garlic (Allium sativum), on the gene expression profiles of peripheral blood mononuclear cells from healthy donors was analyzed. DNA microarray which can detect expression signal of 862 genes revealed that allicin induced the expression of cytokine, chemokine, and immune-related genes in peripheral blood mononuclear cells. In contrast, allicin repressed the expression of adaptive immune-related genes, which are expressed in T helper I lymphocytes. Simultaneous inhibitory and stimulatory effects of allicin were found on inflammatory cells. It is likely that allicin down-regulated the expression of specific genes that were previously up-regulated in resting cells, suggesting a new mechanism by which they exert positive and negative effect. Considering the broad and renewed interest in allicin, the profiles we describe here will be useful in designing more specific and efficient treatment strategies.
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디엠프리(녹차 추출물)가 나균 감염 중간엽 줄기세포의 유전자 발현에 미치는 영향
박란숙,Park, Ran-Sook 한국식품영양학회 2014 韓國食品營養學會誌 Vol.27 No.2
This study found antibacterial activity of $DMfree^{(R)}$ [green tea extract] on facultative bacteria by direct petri dish method and gene array of obligatory M. leprae infected mesenchymal stem cells (MSC). While DMfree showed DPPH radical scavenging effect and high contents of polyphenol, it did not inhibit growth of facultative bacteria such as E. coli and S. aureus on the petri dish. The result does not exclude a possible antibacterial effect of organic solvent extract of green tea rather than DMfree which comes from the water extract of green tea. Pre-treatment of DMfree appeared to have no effect on copy number of 14 genes compared with control MSC by real-time RT-PCR. However pre-treatment of DMfree on M. leprae infected MSC revealed a significant decrease of anti-inflammatory cytokine (IL-6), (P<0.038) and sharp down-regulation of pro-inflammatory cytokine (IL-1). Enhanced expression of VEGFR-1 mRNA was noted in DMfree pretreated M. leprae infected MSC group (P<0.003). These results show that DMfree would stabilize M. leprae infected MSC from further inflammation by down-regulating anti-inflammatory cytokine (IL-6) and pro-inflammatory cytokine (IL-$1{\beta}$). This is the first report on DMfree inhibition of IL-6 and IL-$1{\beta}$ expression in M. leprae infected MSC. Further experiments that detect protein levels of IL-$1{\beta}$ and IL-6 may support the result of this gene array.
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