• 한국약용작물학회지
• /
• 제10권2호
• /
• pp.109-115
• /
• 2002

황칠나무 잎의 추출물의 면역활성증진 실험에서 인간정상 간세포의 경우 모든 추출물이 1.0 Mg/ml의 농도에서 최고 26%이하의 세포독성을 나타내었다. 서로 다른 4가지의 암세포주(MCF7, A549, Hep3B, AGS)에서 50%이상의 저해율을 나타냈고, 정상 세포의 결과와 암세포의 저해율을 비로 나타낸 selectivity의 측정에서 모든 암세포주가 1.5이상의 사멸도를 나타내었고 전체적으로 에탄올 추출물의 효과가 가장 좋았다. 에탄올 추출물의 경우에서 인감 유방암 세포주(MCF7)와 인간 간암세포주(Hep3B)의 경우에서 1.0 mg/ml농도에서 각각 65%와 67%의 저해율을 기록했다. 면역세포 실험에서 에탄올 추출물이 1.0 mg/ml의 농도에서 B세포는 1.22배, T세포는 1.27배의 촉진 활성을 보였고, 6일 동안측정한 $cytokines(IL-6,\;TNF-{\alpha})$의 양도 에탄올 추출물의 경우 T cell의 경우 IL-6은 94pg/m1, $TNF-{\alpha}$은 75pg/ml로 증가하는 것을 알 수 있었다. 이상의 결과로 볼 때 추출 열과 추출용매 등에 의해 황칠나무 잎에 존재하는 여러 가지 유용성분들이 풍부하게 존재하며, crude추출물 중에 에탄올 추출물이 면역활성에서 좋은 효과를 보였다는 것을 알 수가 있었고, 이를 통해서 황칠나무 잎을 이용한 기능성 식품으로의 연구 개발이 통해서 충분히 그 가치가 있다는 것을 알 수 있었다.

• 미생물학회지
• /
• 제40권4호
• /
• pp.334-341
• /
• 2004

와송으로부터 고온 고압의 열수 추출로 다당체 조추출물(OJP1)을 얻어 디스크 확산법, fluorescein diacetate(PDA)법 및 액체 배지 희석법을 통하여 세균과 진균에 대한 항균 활성을 조사하였다. 또한 Sephadex G-50을 통하여 다당체(FI)와 올리고당류(FII)를 분리하고, 이들의 항암 활성을 조사하였다. FI과 FII의 분자량은 각각 30$\~$50 kDa과 1$\~$3 kDa으로 추정되었다. OJP1의 항균 효과는 디스크 확산법에서 Candida albicans가 $20\pm4.9\;mm$의 억제대로 가장 높았으며, Salmonella typhimurium과 Staphylococcus aureus도 각각 $18\pm2.0\;mm$ 와 $17\pm1.0\;mm$의 억제대로 비교적 높게 나타났다. 또한 Escherichia coli와 Pseudomonas aeruginosa에 대해서도 양성 대조군인 프로폴리스보다 높게 나타났으며, FDA법, 액체배지 희석법에서도 비슷한 양상을 나타냈다. OJP1과 Fl, FII의 인체 암세포주와 Sarcoma 180 세포주에 대한 항암 활성을 측정해 본 결과, 이들은 모두 MTT assay와 형태 변화 관찰에서 강력한 암세포주 증식 억제 효과를 나타내었는데, 특히 폐암 세포주인 A549 세포와 자궁 경부암 세포주인 HeLa 세포, 위암 세포주인 AGS에서 현저한 효과를 보였다. 더 나아가 DNA 분절화를 통해 apoptosis 유발 여부를 확인해 본 결과, 대조군에 비해서 이들 물질을 처리한 실험군에서 apoptosis발생을 뜻하는 DNA 분절화 현상이 뚜fut하게 나타났다. 요컨대, OJP1과 Fl, FII는 광범위한 병원성 균주에 대하여 효과적인 항균 활성을 보였으며, 각종 암세포주에 대하여 현저한 항암 활성을 나타내는 것으로 확인되었다.

• 한국식품위생안전성학회지
• /
• 제38권6호
• /
• pp.551-556
• /
• 2023

본 연구는 in vtiro 상에서 부위별 염소고기 열수 추출물의 항산화 활성을 측정하고 추출물 처리에 따른 암세포 주의 세포자멸사 관련 단백질 발현 수준의 변화를 확인하였다. 부위별 염소고기 열수 추출물의 경우, 앞다리, 뒷다리, 등심 및 갈비 부위의 분할육으로 제조하였다. 제조된 열수추출물 중 앞다리와 뒷다리 부위 추출물의 ABTS 라디칼 소거능이 다른 두 부위 추출물보다 유의적으로 높게 나타났다(P<0.05). BAX, p53, p21의 단백질 발현량은 부위에 따른 염소고기 열수추출물 처리 시 AGS 세포주 및 HT-29 세포주에서 모두 비슷한 수준으로 나타났다. 단지, 등심 부위 열수 추출물 처리 시 HT-29 세포주의 p53 발현량이 대조군과 비교하여 유의적으로 높았다(P<0.05). 본 연구 결과는 염소고기 열수 추출물의 항산화 활성 및 일부 세포자멸사 관련 단백질 발현량 변화가 부위에 따라 다르게 나타남을 보이고 있으나 본 결과는 in vitro에서만 수행된 것으로써 결과의 적용 또는 일반화에 한계가 있기 때문에 in vivo에서 후속 연구가 필요하다.

• 한국약용작물학회지
• /
• 제15권3호
• /
• pp.170-176
• /
• 2007

B cell과 T cell을 이용한 면역세포의 생육 촉진 실험에서, B${\cdot}$T cell은 배양 6일째 수피 추출물이 각각 $4.2{\times}10^4$cells/ml, $5.0{\times}10^4$cells/ml로 가장 높은 세포농도를 나타냈다. 면역세포를 이용한 cytokine의 분비량 측정실험에서는 수피추출물이 배양 시간에 따른 cytokine의 분비가 가장 높게 나타나는 것을 확인할 수 있었다. 각 세포의 TNF-${\alpha}$의 분비량은 6일째 최고의 분비량을 나타내었고, 수피 추출물의 경우 대조군에 비해 7-8배 가량 더 높은 분비량을 나타낸 것을 확인하였다. IL-6의 경우 6일째 최고의 분비량을 나타내었고, 수피 추출물에서 가장 높은 분비량을 나타내었다. 또한 때죽나무 추출물을 첨가한 배지에 의한 NK cell의 면역활성을 측정하였으며, 모든 추출물에서 배양시간에 따라 유의적으로 증가하는 것을 확인하였다. 그 중 때죽나무 수피 추출물에서 B cell의 경우 $14.1{\times}10^4$cells/ml, T cell의 경우 $15.3{\times}10^4$cells/ml 으로 가장 높은 생육 활성을 나타내었다. 인간 정상 신장세포인 HEK293을 이용한 세포 독성을 살펴본 결과, 각 추출물은 1.0mg/ml의 농도에서 최고 27.4%의 세포독성을 나타내었다. 또한 항암활성 효과를 살펴보기 위하여 2가지의 암세포를 이용하였는데, AGS와 MCF-7에서 뿌리추출물의 경우 1.0mg/ml의 농도에서 65정도의 높은 억제활성을 나타내었고, 또한 암세포의 생육활성에 대한 정상 세포의 세포독성의 비로 나타낸 선택적 사멸도는 고농도에서는 모두 1.5이상으로 나타나 모두 암세포에 대한 선택성이 있는 것으로 나타났다. Bcl-2 단백질 정량을 통한 항암활성 실험에서는, IOD 값이 수피, 목부, 잎 순으로 각각 72, 186, 200의 결과를 나타내었다. 본 실험 결과를 통하여 때죽나무의 기능성 소재로서의 가능성을 엿볼 수 있었고, 유용성분 분리와 생리활성에 대한 연구가 보다 다각적으로 이루어져야 할 것으로 생각된다.

• Journal of Microbiology and Biotechnology
• /
• 제18권8호
• /
• pp.1431-1438
• /
• 2008

Immunomodulating activities of water-soluble exopolysaccharides (LL-EX) obtained from submerged mycelial culture of Lentinus lepideus were studied and their effectiveness was compared with lipopolysaccharide (LPS). The influence of the LL-EX on macrophage cellular lysosomal enzyme activity was to stimulate up to 267%, 392%, and 464% at the level of 10, 50, and $100{\mu}g/ml$, respectively. When the LL-EX was further fractionated into LL-Fr.I and Fr.II by Sepharose CL-6B gel chromatography, the cellular lysosomal enzyme activity of LL-Fr.II (2.1-fold) was higher than Fr.I (1.2-fold). Moreover, both LL-Fr.I and Fr.II stimulated the cytokines IL-1$\beta$, TNF-$\alpha$, and IL-6 in macrophages. In mixed lymphocyte reaction, LL-Fr.I and Fr.II enhanced the splenocyte proliferation up to 1.2-fold and 1.4-fold ($50{\mu}g/ml$), respectively, stimulating only T lymphocytes. The fractions of LL-EX not show any direct toxicity against human gastric adenocarcinoma cell (AGS). The molecular masses of LL-Fr.I and Fr.II were estimated to be about 1,986 kDa and 21 kDa, respectively. The total sugar and protein contents of the two fractions were 84.97% and 69.88%, and 15.03% and 30.12%, respectively. The sugar and amino acid compositions of the LL-Fr.I and Fr.II were also analyzed in detail.

• 한국식품영양학회지
• /
• 제27권5호
• /
• pp.837-844
• /
• 2014

This study was carried out to analyze the differences in p-hydroxylbenzyl alcohol (HBA) content, antitumor and anti-obesity activities and tyrosinase inhibitory activity between non-fermented G. elata (NFGP) and fermented G. elata powder. The HBA content, which is an index-component of G. elata decreased from 1.58 mg/g before fermentation to 1.07, 0.32, and 0.13 mg/g after the $1^{st}$ fermentation ($1^{st}$ FGP), $2^{nd}$ fermentation ($2^{nd}$ FGP) and $3^{rd}$ fermentation ($3^{rd}$ FGP), respectively. The anti-proliferation effects on the cell lines HT29 and AGS were significantly higher for the fermented G. elata than the NFGP. The antitumor activity was also increased in a fermentation number-dependent manner. During adipocyte differentiation, the ethanol extract of the $3^{rd}$ FGP inhibited lipid accumulation in 3T3-L1 cells significantly better than NFGP and the $1^{st}$ FGP, treated at the concentration of $10{\mu}g/mL$. The tyrosinase inhibitory activity of the $2^{nd}$ FGP at $600{\mu}g/mL$ over was higher than that of kojic acid. At the concentration of $1,000{\mu}g/mL$, the tyrosinase inhibitory activity was increased in a fermentation number-dependent manner. From these results, the fermented G. elata, especially the $3^{rd}$ FGP, is expected to be good candidate for the development of functional food and agents with antitumor, anti-obesity, and tyrosinase inhibitory potential.

• 대한한의학회지
• /
• 제24권2호
• /
• pp.138-147
• /
• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• 한국식품조리과학회지
• /
• 제28권2호
• /
• pp.175-182
• /
• 2012

In this study, antibacterial activity and cytotoxicity of the extracts from pericarp and seed of $Vitis$ $coignetiea$, which were extracted with 0.1% HCl-60% ethanol, were analyzed. The antibacterial activity of the extracts was determined by paper disc diffusion method against food spoilage and food-borne pathogens. The pericarp extract showed high antibacterial activity against $Bacillus$ $cereus$, $Escherichia$ $coli$ O157:H7, and $Pseudomonas$ $aeruginosa$, and the seed extract represented the high antibacterial activity against $B.$ $cereus$, $E.$ $coli$ O157:H7, and $Staphylococcus$ $aureus$. The cytotoxicity of the $Vitis$ $coignetiea$ extract against human cancer cells was determined using the MTT assay and SRB assay. The pericarp extract represented strong growth-inhibition activity against G361 and Hep3B cells and the seed extract greatly inhibited the growth of HeLa and G361 cells in the MTT assay. In addition, the pericarp extract displayed a high inhibition activity against the growth of AGS cells and the seed extract greatly inhibited the growth of HeLa, Hep3B, and MCF7 cells in the SRB assay. Especially, the cytotoxicities of the seed extract against HeLa were significantly higher than those of the extract against other cancer cells at all test concentrations. This study demonstrates that the extract from pericarp and seed of $Vitis$ $coignetiea$ possess high antibacterial activity and cytotoxicity.

• BMB Reports
• /
• 제55권7호
• /
• pp.348-353
• /
• 2022

Gastrointestinal cancer is associated with a high mortality rate. Here, we report that the splice variant of NRP/B contributes to tumorigenic activity in highly malignant gastric cancer through dissociation from the tumor repressor, HDAC5. NRP/B mRNA expression is significantly higher in the human gastric cancer tissues than in the normal tissues. Further, high levels of both the NRP/B splice variant and Lgr5, but not the full-length protein, are found in highly tumorigenic gastric tumor cells, but not in non-tumorigenic cells. The loss of NRP/B markedly inhibits cell migration and invasion, which reduces tumor formation in vivo. Importantly, the inhibition of alternative splicing increases the levels of NRP/B-1 mRNA and protein in AGS cells. The ectopic expression of full-length NRP/B exhibits tumor-suppressive activity, whereas NRP/B-2 induces the noninvasive human gastric cancer cells tumorigenesis. The splice variant NRP/B-2 which loses the capacity to interact with tumor repressors promoted oncogenic activity, suggesting that the BTB/POZ domain in the N-terminus has a crucial role in the suppression of gastric cancer. Therefore, the regulation of alternative splicing of the NRP/B gene is a potential novel target for the treatment of gastrointestinal cancer.

• Journal of Gastric Cancer
• /
• 제22권4호
• /
• pp.319-338
• /
• 2022

Purpose: Gastric cancer (GC) has high morbidity and mortality, the cure rate of surgical treatment and drug chemotherapy is not ideal. Therefore, development of new treatment strategies is necessary. We aimed to identify the mechanism underlying Sp1 regulation of GC progression. Methods and Methods: The levels of Sp1, β-catenin, SET domain bifurcated 1 (SETDB1), and 15-hydroxyprostaglandin dehydrogenase (HPGD) were detected by quantitative reverse transcription polymerase chain reaction and western blot analysis. The targets of SETDB1 were predicted by AnimalTFDB, and dual-luciferase reporter assay was used for confirming the combination of Sp1, β-catenin, and SETDB1. HGC27 or AGS cells (1×10⁶ cells/mouse) were injected into mice via the caudal vein for GC model establishment. The level of Ki67 was detected using immunohistochemistry, and hematoxylin and eosin staining was performed for evaluating tumor metastasis in mice with GC. Results: HPGD was inhibited, while the protein levels of Sp1, β-catenin, and SETDB1 were up-regulated in GC tissues and cell lines. HPGD overexpression or SETDB1 silencing inhibited the proliferation, invasion, and migration of GC cells, and Sp1 regulated the proliferation, invasion, and migration of GC cells in a β-catenin-dependent manner. Furthermore, HPGD served as a target of SETDB1, and it was negatively regulated by SETDB1; additionally, Sp1 and β-catenin bound to the SETDB1 promoter and negatively regulated HPGD expression. We proved that Sp1 regulated GC progression via the SETDB1/HPGD axis. Conclusions: Our findings revealed that Sp1 transcriptionally inhibited HPGD via SETDB1 in a β-catenin-dependent manner and promoted the proliferation and metastasis of GC cells.