• Journal of Plant Biotechnology
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• 제2권2호
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• pp.55-60
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• 2000

Experiments initiated 30 years ago to obtain selectable markers have led to a series of studies of Trp biosynthesis and anthranilate synthase (AS) the control enzyme using largely plant tissue cultures since they have experimental properties that can be readily exploited. Enzymological and compound feeding studies provided evidence that AS is the control point in the Trp biosynthesis branch and that altering the AS feedback control by the selection of mutants resistant to the Trp analog 5-methyl-tryptophan (5MT) can lead to the overproduction of this important amino acid. Plants regenerated from these Trp overproducing lines of most species also had high free Trp levels but Nicotiana tabaum (tobacco) plants expressed the feedback altered AS only in cultured cells and not in the regenerated plants. further tests by transient and stable expression of the cloned promoter for the naturally occurring tobacco feedback-insensitive AS, denoted ASA2, confirmed the tissue culture specific nature of the expression control. The 5MT caused by the expression of a feedback-insensitive AS from tobacco has been used to select protoplast fusion hybrids with several species since the resistance is expressed dominantly. Recently the ASA2 gene has been used successfully as a selectable marker to select transformed Astragalus sinicus and Glycine max hairy roots induced by Agrobactetium rhizogenes. These results show that the ASA2y-subunit can interact with the y-subunit of another species to form active feedback-insensitive enzyme that may be useful for selecting transformed cells. Plastid DNA transformation of tobacco has also effectively expressed ASA2 in the compartment in which Trp biosynthesis is localized in the cell.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7603-7609
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• 2014

Glutathione is a thiol compound that plays an important role in the antioxidant defense system of the cell and its deficiency leads to an increased susceptibility to oxidative stress and, thus, progression of many disease states including head and neck cancer. In the present study, alterations of glutathione levels were investigated in study cohort of 500 samples (cohort 1 containing 200 head and neck cancer blood samples along with 200 healthy controls and cohort II with 50 head and neck squamous cell carcinoma tissue samples along with 50 control tissues) by high performance liquid chromatography. The results indicated that mean blood glutathione levels were significantly reduced in head and neck cancer patients (p<0.001) compared to respective controls. In contrast, the levels of glutathione total (p<0.05) and glutathione reduced (p<0.05) were significantly elevated in head and neck squamous cell carcinoma tissues compared to the adjacent cancer-free control tissues. In addition to this, pearson correlation performed to correlate different tissue glutathione levels (GSH) with clinical/pathological parameters demonstrated a significant negative correlation between pT-stage and GSH level ($r=-0.263^{**}$; p<0.01), C-stage and GSH level ($r=-0.335^{**}$; p<0.01), grade and GSH ($r=-0.329^{**}$; p<0.01) and grade versus redox index ($r=-0.213^{**}$; p<0.01) in HNSCC tissues. Our study suggests that dysregulation of glutathione levels in head and neck cancer has the potential to predict metastasis, and may serve as a prognostic marker.

• Journal of Ginseng Research
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• 제42권1호
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• pp.68-74
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• 2018

Background: Ginsenosides have been reported to have many health benefits, including anti-inflammatory effects, and the resolution of inflammation is now considered to be an active process driven by M2-type macrophages. In order to determine whether ginsenosides modulate macrophage phenotypes to reduce inflammation, 11 ginsenosides were studied with respect to macrophage polarization and the resolution of inflammation. Methods: Mouse peritoneal macrophages were polarized into M1 or M2 phenotypes. Reverse transcription-polymerase chain reaction, Western blotting, and measurement of nitric oxide (NO) and prostaglandin $E_2$ levels were performed in vitro and in a zymosan-induced peritonitis C57BL/6 mouse model. Results: Ginsenoside $Rg_3$ was identified as a proresolving ginseng compound based on the induction of M2 macrophage polarization. Ginsenoside $Rg_3$ not only induced the expression of arginase-1 (a representative M2 marker gene), but also suppressed M1 marker genes, such as inducible NO synthase, and NO levels. The proresolving activity of ginsenoside $Rg_3$ was also observed in vivo in a zymosan-induced peritonitis model. Ginsenoside $Rg_3$ accelerated the resolution process when administered at peak inflammatory response into the peritoneal cavity. Conclusion: These results suggest that ginsenoside $Rg_3$ induces the M2 polarization of macrophages and accelerates the resolution of inflammation. This finding opens a new avenue in ginseng pharmacology.

• Journal of Ginseng Research
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• 제38권2호
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• pp.89-96
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• 2014

The protective effect of ginsenoside Re, isolated from ginseng berry, against acute gastric mucosal lesions was examined in rats with a single intraperitoneal injection of compound 48/80 (C48/80). Ginsenoside Re (20 mg/kg or 100 mg/kg) was orally administered 0.5 h prior to C48/80 treatment. Ginsenoside Re dose-dependently prevented gastric mucosal lesion development 3 h after C48/80 treatment. Increases in the activities of myeloperoxidase (MPO; an index of neutrophil infiltration) and xanthine oxidase (XO) and the content of thiobarbituric acid reactive substances (TBARS; an index of lipid peroxidation) and decreases in the contents of hexosamine (a marker of gastric mucus) and adherent mucus, which occurred in gastric mucosal tissues after C48/80 treatment, were significantly attenuated by ginsenoside Re. The elevation of Bax expression and the decrease in Bcl2 expression after C48/80 treatment were also attenuated by ginsenoside Re. Ginsenoside Re significantly attenuated all these changes 3 h after C48/80 treatment. These results indicate that orally administered ginsenoside Re protects against C48/80-induced acute gastric mucosal lesions in rats, possibly through its stimulatory action on gastric mucus synthesis and secretion, its inhibitory action on neutrophil infiltration, and enhanced lipid peroxidation in the gastric mucosal tissue.

• 한국식품과학회지
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• 제41권6호
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• pp.717-721
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• 2009

고장초 메탄올 추출물의 항알레르기 활성을 조사하기 위해, 비만세포주인 RBL-2H3 세포를 이용하였으며, 고장초 추출물을 처리한 후 IgE-항체, A23187 및 PMA를 이용하여 세포를 활성화시켜 분비되는 $\beta$-hexosaminidase, TNF-$\alpha$의 양을 측정하였다. 그 결과 고장초 추출물이 활성화된 비만세포에서 분비되는 $\beta$-hexosaminidase, TNF-$\alpha$를 농도 의존적으로 억제하는 것을 확인할 수 있었다. 또한, 비만세포에 탈과립이 발생할 경우 나타나는 형태학적 변화에 대한 시료의 영향을 검색하기 위해 고장초 추출물을 처리한 세포에 compound 48/80을 처리하였으며, 그 결과 고장초 추출물을 처리한 경우 대조군에 비해 탈과립이 현저히 억제되는 것을 관찰하였다. 따라서 고장초 추출물은 다양한 자극으로 인한 비만세포의 탈과립 저해를 통해 알레르기 반응을 억제시키는 것으로 판단되며, 본 연구결과는 고장초의 항알레르기 기능성 소재로의 이용 가능성을 시사해준다.

• 생약학회지
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• 제34권2호통권133호
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• pp.123-127
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• 2003

Atrartylodis Rhizoma(蒼朮)'s origin plants are Atratylodes lancea and A. Chinensis in Chinese, Japanese and Korean pharmacopoeia. A. Japonica is only indigenous in Korea, it is actually used as Atractylodis Rhizoma in Korean market. A. lancea is used in Hunan province, China and A. Chinensis is used in Hubei province, China. It is impossible to distinguish with species differency as macro- and micro-morphology. We tried to distinguish with species differency by HPLC and GC-Mass spectra. Atractylone(mw. 216) which is a marker compound in Atractylodis Rhizoma Alba(白朮) was detected in A. japonica. Atractylodin (mw.182) was detected in A. lancea and two eudesmadien derivatives (mw. 204) were detected in A. chinensis. HPLC chromatogram showed the same patterns. As a result, we propose that A. japonica will be added as Atractylodis Rhizoma (蒼朮)'s origin plant in Korean Pharmacopoeia. Atractylodis Rhizoma Alba(白朮)'s origin plants are A. macrocephala in China, and A. Japonica and A. ovata in Korea and Japan. In GC-Mass analysis, all samples showed same patterns and the main compound was atractylone.

• Natural Product Sciences
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• 제26권3호
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• pp.230-235
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• 2020

A pregnane steroid, 3α-hydroxy-pregn-7-ene-6,20-dione (1), was isolated from a Hydractinia-associated Cladosporium sphaerospermum SW67 by repetitive column chromatographic separation and high-performance liquid chromatography (HPLC) purification. The planar structure of 1 was elucidated from the analysis of the spectroscopic data (1D and 2D NMR spectra) and LC-MS data. The absolute configuration of 1 was determined by interpretation of ROESY spectrum of 1, together with the comparison of reported spectroscopic values in previous studies. To the best of our knowledge, this is the first report of the identification of the pregnane scaffold from C. sphaerospermum, a natural source. Compound 1 was evaluated for its effects on lipid metabolism and adipogenesis during adipocyte maturation and showed that compound 1 substantially inhibited lipid accumulation compared to the control. Consistently, the expression of the adipocyte marker gene (Adipsin) was reduced upon incubation with 1. Further, we evaluated the effects of 1 on lipid metabolism by measuring the transcription of lipolytic and lipogenic genes. The expression of the lipolytic gene ATGL was significantly elevated upon exposure to 1 during adipogenesis, whereas the expression of lipogenic genes FASN and SREBP1 was significantly reduced upon treatment with 1. Thus, our findings provide experimental evidence that the steroid derived from Hydractinia-associated C. sphaerospermum SW67 is a potential therapeutic agent for obesity.

• Nutrition Research and Practice
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• 제14권5호
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• pp.478-489
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• 2020

BACKGROUND/OBJECTIVES: Morusin, a marker component of Morus alba L., possesses anti-cancer activity. The objective of this study was to determine autophagy-inducing effect of morusin in non-small cell lung cancer (NSCLC) cells and investigate the underlying mechanism. SUBJECTS/METHODS: Autophagy induction and the expression of autophagy-related proteins were analyzed by LC3 immunofluorescence and western blot, respectively. The role of autophagy and AMP-activated protein kinase (AMPK) was determined by treating NSCLC cells with bafilomycin A1, an autophagy inhibitor, and compound C, an AMPK inhibitor. Cytotoxicity and apoptosis induction were determined by MTT assay, trypan blue exclusion assay, annexin V-propidium iodide (PI) double staining assay, and cell cycle analysis. RESULTS: Morusin increased the formation of LC3 puncta in the cytoplasm and upregulated the expression of autophagy-related 5 (Atg5), Atg12, beclin-1, and LC3II in NSCLC cells, demonstrating that morusin could induce autophagy. Treatment with bafilomycin A1 markedly reduced cell viability but increased proportions of sub-G1 phase cells and annexin V-positive cells in H460 cells. These results indicate that morusin can trigger autophagy in NSCLC cells as a defense mechanism against morusin-induced apoptosis. Furthermore, we found that AMPK and its downstream acetyl-CoA carboxylase (ACC) were phosphorylated, while mammalian target of rapamycin (mTOR) and its downstream p70S6 kinase (p70S6K) were dephosphorylated by morusin. Morusin-induced apoptosis was significantly increased by treatment with compound C in H460 cells. These results suggest that morusin-induced AMPK activation could protect NSCLC cells from apoptosis probably by inducing autophagy. CONCLUSIONS: Our findings suggest that combination treatment with morusin and autophagy inhibitor or AMPK inhibitor might enhance the clinical efficacy of morusin for NSCLC.

• 한국식품과학회지
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• 제45권1호
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• pp.20-24
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• 2013

아몬드와 땅콩의 비조사 시료와 1, 3, 5, 10 kGy로 전자선을 조사한 시료에서 유도된 hydrocarbon류의 생성량을 분석하기 위하여 지방을 n-hexane으로 추출하여 florisil column으로 분리 및 GC/MS로 분석하였다. 전자선 조사된 아몬드와 땅콩에서 유래한 hydrocarbon류는 oleic acid에서 1,7-hexadecadiene($C_{16:2}$)과 8-heptadecene($C_{17:1}$), linoleic acid에서 1,7,10-hexadecatriene($C_{16:3}$)과 6,9-heptadecadiene($C_{17:2}$), 그리고 palmitic acid에서 1-tetradecene($C_{14:1}$) 및 pentadecane($C_{15:0}$) 생성되었고, oleic acid와 palmitic acid에서는 $C_{n-2}$, linoleic acid에서는 $C_{n-1}$ 화합물이 보다 더 높게 생성되었다. 전자선을 1, 3, 5, 10 kGy의 선량별로 조사한 시료에서 생성된 hydrocarbon류의 함량은 조사선량에 따라 증가하였고 비조사 시료에서는 확인되지 않았다. 특히, oleic acid와 linoleic acid에서 유래한 1,7-hexadecadiene($C_{16:2}$), 8-heptadecene($C_{17:1}$)과 1,7,10-hexadecatriene($C_{16:3}$) 및 6,9-heptadecadiene($C_{17:2}$)의 경우 아몬드와 땅콩 모두에서 다른 hydrocarbon류에 비해 전자선 조사에 의한 생성량이 높게 나타난바 전자선 조사유무 판별 marker로 사용 가능함을 확인하였다.

• 한국식품영양과학회지
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• 제31권6호
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• pp.958-964
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• 2002

방사선 조사된 닭고기에서 휘발성 조사물질을 선정하기 위하여 각 선량별(0,1,3,5,10 kGy)로 조사된 닭고기를 시료로 하여 P&T법으로 휘발성 성분을 추출한 후 GC/MS법으로 분석하였으며, 동시에 LLCE법의 단점을 보완한 상보적인 분석방법으로서의 가능성을 제시하고자 하였다 그 결과 P&T/GC/MS법에 의해 총 119종의 휘발성 성분이 검출되었으며 이는 알데히드류(7종), 케톤류(22종), 알콜류(8종), 에스테르화합물류(30종), 탄화수소류(36종), 방향족화합물류(8종) 및 기타 화합물류(8종)로 구성되어 있었다. 이중 21종의 화합물이 LLCE법과 공통적으로 검출되었고, 나머지 98종은 P&T법 에 의해서만 검출됨으로서 LLCE법과 P&T법을 동시에 사용할 경우 더욱 폭넓은 휘발성 향기성분의 분석이 가능하였다. 그리고 P&T법에 의해 추출된 휘발성 향기 성분중 방사선 조사선량과 함량간의 회귀분석 및 상관분석을 행한 결과 hexene, propanol 및 1,3 bis(1,1-dimethylethy)benzene 등 3종의 휘발성 화합물이 유의적인(p<0.01 또는 p<0.05) 양의 상관성 (r)0.90)을 나타내어 P&T법에 의한 닭고기의 방사선 조사 판별을 위한 표지물질로 선정되었다.