• Title/Summary/Keyword: human intestinal epithelial cells

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Sulfoquinovosylmonoacylglycerols regulating intestinal inflammation in co-culture system from the brown alga Turbinaria ornata

  • Lee, Seon Min;Kim, Na-Hyun;Ji, Yeong Kwang;Kim, Yun Na;Jeon, You-Jin;Heo, Jeong Doo;Jeong, Eun Ju;Rho, Jung-Rae
    • ALGAE
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    • v.35 no.2
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    • pp.201-212
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    • 2020
  • The inflammatory bowel diseases (IBD) including ulcerative colitis and Crohn disease are characterized by chronic inflammation throughout the gastrointestinal tract. The prevalence of IBD has been increasing worldwide, and has sometimes led to irreversible impairment of gastrointestinal structure and functions. In the present study, we identified a new sulfoquinovosylmonoacylglycerols (SQMG) (1) together with two known SQMGs (2 and 3) regulating intestinal inflammation from the brown alga Turbinaria ornata. The anti-inflammatory properties of two bioactive SQMGs, 1 and 2 were evaluated using an in vitro co-culture system consisting of human epithelial Caco-2 cells and PMA (phorbol 12-myristate 12-acetate)-differentiated THP-1 macrophages. Treatment with 1 or 2 inhibited the production nitric oxide and prostaglandin E2 induced by lipopolysaccharide and interferon γ challenge. The expressions of inducible nitric oxide synthase and cyclooxygenase 2 were markedly down-regulated in response to inhibition of nuclear factor κB translocation to nucleus. These findings suggest the potential use of the brown alga T. ornata and its biologically active metabolites SQMGs as pharmaceutical adjuvants in the treatment of inflammation-related diseases, including IBD.

Immune-triggering effect of the foodborne parasite Kudoa septempunctata through the C-type lectin Mincle in HT29 cells

  • Shin, Ji-Hun;Yang, Jung-Pyo;Seo, Seung-Hwan;Kim, Sang-Gyun;Kim, Eun-Min;Ham, Do-Won;Shin, Eun-Hee
    • BMB Reports
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    • v.53 no.9
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    • pp.478-483
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    • 2020
  • Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

Mechanism of Apoptosis & Tumor Growth Inhibition of Agrimonia pilosa Ledebour(APL) in vitro and in vivo (선학초(짚신나물)에 의한 in vitro와 in vivo에서의 암세포사멸 기전 탐색)

  • Choi, Soon-Ja;Baik, Jong-Woo;Park, Jong-Hyeong;Jun, Chan-Yong;Choi, You-Kyung;Ko, Seung-Gyu
    • The Journal of Internal Korean Medicine
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    • v.30 no.2
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    • pp.399-409
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    • 2009
  • Objectives : The aim of this study was to experiment the antitumor activity of Agrimonia pilosa Ledebour (APL) in human stomach cancer (AGS) cell lines (in vitro) and male C57BL/6J mouse (in vivo). Methods : The effects of the ethanol extract from the plant on several transplantable rodent tumors were investigated in vitro by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. DNA content analysis and Western blot analysis. Agrimonia pilosa Ledebour (APL) was given to rats with Lewis Lung Carcinoma (LLC) cells. The experimental rats were divided into 3 groups in vivo. Saline was injected into the abdominal cavity in the first group, 50 mg/kg APL was injected into the abdominal cavity in the second group and 100 mg/kg was injected into the abdominal cavity in the third group. After that, we checked their tumor volume periodically. Results : At first, human gastric cancer (AGS) cell lines (in vitro) showed decreased cell viability, and increased $sub-G_1$ contents. When we experimented rat intestinal epithelial (RIE)l as same condition, this result didn't show. With this, compared to normal cells, Agrimonia pilosa Ledebour (APL) led selectively to the extinction of cells only in human gastric cancer. Moreover, we showed that the traditional herbal medicine APL induced caspase-dependent apoptosis in AGS cells. Next, APL inhibited the growth of LLC-bearing mouse tumor. However, we could not verify APL induced caspase-dependent apoptosis in LLC-bearing mouse tumor. Conclusions : The roots of Agrimonia pilosa Ledebour (APL) contain some antitumor constituents.

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Effect of Baegieum(BGU) on Oxidant induced cell death in human intestinal epithelial cells (배기음(排氣飮)이 인간(人間)의 장관(腸管) 상피세포(上皮細胞)에서 Oxidant에 의해 유발(誘發)된 세포사망(細胞死亡)과 DNA 손상(損傷)에 미치는 영향)

  • Kim, Woo-Hwan;Kim, Won-Ill
    • The Journal of Internal Korean Medicine
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    • v.21 no.1
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    • pp.13-22
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    • 2000
  • 목적 : 본(本) 연구(硏究)는 배기음(排氣飮)이 인간(人間)의 장관내(腸管內)에서 산화물(酸化物)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡) 및 DNA의 손상(損傷)을 방지할수 있는지를 검증(檢證)하기 위한 실험(實驗)이다. 방법 : 배양(培養)된 인체장관(人體腸管) 세포계열(細胞系列)인 Caco-2 세포(細胞)에서 세포(細胞)의 사망(死亡)은 trypan bile의 소실정도에 의해서 평가했으며, DNA의 손상(損傷)은 double stranded DNA의 파괴정도를 측정하여 평가하였다. $H_2O_2$는 표본(標本) 산화제(酸化劑)로 사용되었다. 결과 : $H_2O_2$에 노출된 세포들의 세포사망(細胞死亡) 정도는 노출시간과 용량에 비례하여 증가하는 양상을 보였다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)되는 세포방지를 방지하였고, 0.05-1%의 농도범위에 걸쳐서는 그 효과가 용량에 비례하여 증가하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 세포손상(細胞損傷)은 catalase(hydrogen peroxide scavenger enzyme)와 deferoxamine(iron chelator)에 의해 억제되었다. 그러나 강력한 항산화제(抗酸化劑)인 DPPD는 $H_2O_2$에 의해 유발(誘發)되는 세포손상(細胞損傷)에는 영향을 주지 못했다. $H_2O_2$에 의해 유발(誘發)된 지질(脂質)의 과산화(過酸化)는 배기음(排氣飮)과 DPPD에 의해 억제되었다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상(損傷)은 배기음(排氣飮)에 의해 방지되었으며 용량에 의존하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상은 catalase와 deferoxamine에 의해 억제되었지만 DPPD는 억제시키지 못했다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)된 ATP의 소실을 회복시켰다. 이러한 실험결과 $H_2O_2$에 의해 유발(誘發)된 세포(細胞)의 손상(損傷)은 지질(脂質)의 과산화(過酸化)와는 다른 독립적인 기전에 의해 일어남을 나타낸다. 결론 : 이러한 결과들로 볼 때 Caco-2 세포(細胞)에서 배기음(排氣飮)이 항산화작용(亢酸化作用)보다는 다른 기전을 통하여 Caco-2 세포안에서 산화제(酸化劑)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡)와 DNA의 손상(損傷)을 방지할 수 있다는 것을 가리킨다. 따라서 본 연구(硏究)는 배기음(排氣飮)이 반응성산소기(反應性酸素基)에 의해 매개된 인체(人體) 위장관질환(胃腸管疾患)의 치료(治療)에 사용할 수 있을 가능성(可能性)이 있음을 제시하고 있다.

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Comparative Genomics Approaches to Understanding Virulence and Antimicrobial Resistance of Salmonella Typhimurium ST1539 Isolated from a Poultry Slaughterhouse in Korea

  • Kim, Eunsuk;Park, Soyeon;Cho, Seongbeom;Hahn, Tae-Wook;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.962-972
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    • 2019
  • Non-typhoidal Salmonella (NTS) is one of the most frequent causes of bacterial foodborne illnesses. Considering that the main reservoir of NTS is the intestinal tract of livestock, foods of animal origin are regarded as the main vehicles of Salmonella infection. In particular, poultry colonized with Salmonella Typhimurium (S. Typhimurium), a dominant serotype responsible for human infections, do not exhibit overt signs and symptoms, thereby posing a potential health risk to humans. In this study, comparative genomics approaches were applied to two S. Typhimurium strains, ST1539 and ST1120, isolated from a duck slaughterhouse and a pig farm, respectively, to characterize their virulence and antimicrobial resistance-associated genomic determinants. ST1539 containing a chromosome (4,905,039 bp; 4,403 CDSs) and a plasmid (93,876 bp; 96 CDSs) was phylogenetically distinct from other S. Typhimurium strains such as ST1120 and LT2. Compared to the ST1120 genome (previously deposited in GenBank; CP021909.1 and CP021910.1), ST1539 possesses more virulence determinants, including ST64B prophage, plasmid spv operon encoding virulence factors, genes encoding SseJ effector, Rck invasin, and biofilm-forming factors (bcf operon and pefAB). In accordance with the in silico prediction, ST1539 exhibited higher cytotoxicity against epithelial cells, better survival inside macrophage cells, and faster mice-killing activity than ST1120. However, ST1539 showed less resistance against antibiotics than ST1120, which may be attributed to the multiple resistanceassociated genes in the ST1120 chromosome. The accumulation of comparative genomics data on S. Typhimurium isolates from livestock would enrich our understanding of strategies Salmonella employs to adapt to diverse host animals.

Gastrointestinal Carriage of Escherichia coli with Hemagglutination Activity and Colonization Factor Antigen I and its Relation to Diarrhea (Mannose-resistant Hemagglutination(MRHA) 및 Colonization Factor Antigen I(CFA I)을 표현하는 대장균의 장관내 존재와 설사증 발현과의 관계)

  • Ro, Sung-Hyun;Kim, Kyung-Hee;Cho, Yaug-Ja;Suh, Inn-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.147-153
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    • 1987
  • Colonization factor antigen I(CFA I) has been shown to be one of several virulence factors that promote attachment of enterotoxigenic E. coli(ETEC) to small intestinal epithelial cells of humans. The ability of ETEC to produce mannose-resistant hemagglutination(MRHA) of human blood group A has been used to detect CFA I. To determine gastrointestinal carriage in Korean children of E. coli with MRHA and CFA I, 116 strains of E. coli from diarrheal children admitted to Hanyang University Hospital were examined for MRHA of human erythrocytes and the presence of CFA I. Of 45 ETEC strains, 18(40%) gave a positive MRHA($MRHA^+$) and eight(18%) were positive for CFA I(CFA $I^+$). ETEC with CFA I were all heat-stable enterotoxin(ST) producers and two of these strains were of serogroups $O_{25}$. Of 17 classic enteropathogenic E. coli(EPEC), 7(41%) were $MRHA^+$ but all were negative for CFA I(CFA $I^-$). Of 30 enteroadherent E. coli(EAEC) strains, 11(37%) were $MRHA^+$ and one was CFA $I^+$. Of 24 nonpathogenic E. coli, 4(17%) were $MRHA^+$ but all were CFA $I^-$. It was shown that MRHA was common in all strains of E. coli, CFA I was limited only to ST producing ETEC and EAEC; although MRHA is a useful screening procedure, serologic tests seem to be necessary to comfirm CFA I production. CFA I was associated with a lower proportion of ETEC isolates in Korea than has been reported for other locations.

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Apoptosis of Kinetin Riboside in Colorectal Cancer Cells Occurs by Promoting β-Catenin Degradation

  • TaeKyung Nam;Wonku Kang;Sangtaek Oh
    • Journal of Microbiology and Biotechnology
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    • v.33 no.9
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    • pp.1206-1212
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    • 2023
  • The Wnt/β-catenin pathway plays essential roles in regulating various cellular behaviors, including proliferation, survival, and differentiation [1-3]. The intracellular β-catenin level, which is regulated by a proteasomal degradation pathway, is critical to Wnt/β-catenin pathway control [4]. Normally, casein kinase 1 (CK1) and glycogen synthase kinase-3β (GSK-3β), which form a complex with the scaffolding protein Axin and the tumor suppressor protein adenomatous polyposis coli (APC), phosphorylate β-catenin at Ser45, Thr41, Ser37, and Ser33 [5, 6]. Phosphorylated β-catenin is ubiquitinated by the β-transducin repeat-containing protein (β-TrCP), an F-box E3 ubiquitin ligase complex, and ubiquitinated β-catenin is degraded via a proteasome pathway [7, 8]. Colorectal cancer is a significant cause of cancer-related deaths worldwide. Abnormal up-regulation of the Wnt/β-catenin pathway is a major pathological event in intestinal epithelial cells during human colorectal cancer oncogenesis [9]. Genetic mutations in the APC gene are observed in familial adenomatous polyposis coli (FAP) and sporadic colorectal cancers [10]. In addition, mutations in the N-terminal phosphorylation motif of the β-catenin gene were found in patients with colorectal cancer [11]. These mutations cause β-catenin to accumulate in the nucleus, where it forms complexes with transcription factors of the T-cell factor/lymphocyte enhancer factor (TCF/LEF) family to stimulate the expression of β-catenin responsive genes, such as c-Myc and cyclin D1, which leads to colorectal tumorigenesis [12-14]. Therefore, downregulating β-catenin response transcription (CRT) is a potential strategy for preventing and treating colorectal cancer. Plant cytokinins are N6-substituted purine derivatives; they promote cell division in plants and regulate developmental pathways. Natural cytokinins are classified as isoprenoid (isopentenyladenine, zeatin, and dihydrozeatin), aromatic (benzyladenine, topolin, and methoxytopolin), or furfural (kinetin and kinetin riboside), depending on their structure [15, 16]. Kinetin riboside was identified in coconut water and is a naturally produced cytokinin that induces apoptosis and exhibits antiproliferative activity in several human cancer cell lines [17]. However, little attention has been paid to kinetin riboside's mode of action. In this study, we show that kinetin riboside exerts its cytotoxic activity against colon cancer cells by suppressing the Wnt/β-catenin pathway and promoting intracellular β-catenin degradation.

Oxidative Stress of Mouse Fed with ${\gamma}$-Irradiated Soybean Diet (대두 함유 방사선 조사식이를 섭취한 Mouse의 산화적 스트레스)

  • Park, Sun-Young;Seo, Dae-Young;Suh, Kwang-Sun;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.40 no.2
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    • pp.138-146
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    • 2007
  • Food irradiation has been steadily increased in many countries concomitantly with increasing international trades. Harmful contaminants naturally occurred from foods which contain high levels of unsaturated fatty acids that are easily oxidized can affect the human anti-oxidation system through the generation of free radicals. Moreover, previous studies proved that ${\gamma}$-irradiation may cause production of free radicals in food. We investigated the effect of ${\gamma}$-irradiated soybeans in relation to oxidative stress in mice. Oxidative index of mice was evaluated by TBARS, DNA fragmentation in various organs such as blood lymphocytes, liver and kidney. Forty male ICR mice were equally divided into 4 groups and fed control diet or ${\gamma}$-irradiated diet containing 50% soybeans (5, 10, and 20 kGy, respectively) for 8 weeks. Peroxide values of the irradiated diets were higher than that of the non-irradiated one and increased according to the storage period. There was no significant difference in weight gain as well as in TBARS value in plasma and kidney of all groups. Liver TBARS value of the group fed with irradiated diet at 20 kGy increased significantly compared with the control group (p < 0.05). DNA oxidative damage as measured by alkaline comet assay showed that % tail DNA in the blood lymphocytes of 5 kGy and 10 kGy groups increased significantly over the control group (p < 0.05). Also, tail moments of 5 kGy and 10 kGy groups were higher than that of the control group. Ultrastructural examination shows myeline figures and swollen mitochondria in parietal and intestinal epithelial cells of the group fed with irradiated diet. Therefore, considering unsaturated fatty acid content, consumption of soybeans ${\gamma}$-irradiated with over 20 kGy or repeatedly may decrease the body's antioxidant mechanism.

Detecting of Periodic Fasciculations of Avian Muscles Using Magnetic and Other Multimedia Devices

  • Nakajima, Isao;Tanaka, Sachie;Mitsuhashi, Kokuryo;Hata, Jun-ichi;Nakajima, Tomo
    • Journal of Multimedia Information System
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    • v.6 no.4
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    • pp.293-302
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    • 2019
  • In the past, there was a theory that influenza wasn't transmitted directly from birds but was infected to humans via swains. Recently, molecular level research has progressed, and it was confirmed that the avian influenza virus can directly infected to human lung and intestinal epithelial cells. Three pandemicsin the past 100 years were also infected to humans directly from birds. In view of such scientific background, we are developing a method for screening sick birds by monitoring the physiological characteristics of birds in a contactless manner with sensors. Here, the movement of respiratory muscles and abdominal muscles under autonomic innervation was monitored using a magnet and Hall sensor sewn on the thoracic wall, and other multimedia devices. This paper presents and discusses the results of experiments involving continuous periodic noise discovered during flight experiments with a data logger mounted on a Japanese pheasant from 2012 to 2015. A brief summary is given as the below: 1. Magnet and Hall sensor sewn to the left and right chest walls, bipolar electrocardiograms between the thoracic walls, posterior thoracic air sac pressure, angular velocity sensors sewn on the back and hips, and optical reflection of LEDs (blue and green) from the skin of the hips allow observation of periodic vibrations(fasciculations) in the waves. No such analysis has been reported before. 2. These fasciculations are presumed to be derived from muscle to maintain and control air sac pressure. 3. Since each muscle fiber is spatially Gaussian distributed from the sympathetic nerve, the envelope is assumed to plot a Gaussian curve. 4. Since avian trunk muscles contract periodically at all time, we assume that the sympathetic nerve dominates in their control. 5. The technique of sewing a magnet to the thoracic wall and measuring the strength of the magnetic field with a Hall sensor can be applied to screen for early stage of avian influenza, with a sensor attached to the chicken enclosure.

Effect of Mokhyangsungi-tang(MHS) on antioxidative ability in human intestinal epithelial cells (목향순기탕(木香順氣湯)이 인간의 장관상피세포내에서 항산화효과에 미치는 영향)

  • Kim, Ji-Woong;Kim, Woo-Hwan;Kim, Won-Ill
    • The Journal of Internal Korean Medicine
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    • v.22 no.3
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    • pp.383-391
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    • 2001
  • 목적 : 본 연구는 목향순기탕(木香順氣湯)이 인간의 장관상피세포 계열인 Caco-2 세포에서 항산화작용을 증진시키는 효과가 있는지 검증하기 위한 실험이다. 방법 : 배양된 인간장관 세포계열인 Caco-2 세포에서 세포의 사망은 trypan blue의 소실정도에 의해 평가했으며 $H_2O_2$는 표본산화제로 사용되었다. 결과 : $H_2O_2$에서 노출된 세포들은 용량에 비례하여 세포 사망하는 결과를 보였다. 목향순기탕(木香順氣湯)은 $H_2O_2$에 의해 유발된 세포사망을 방지하였고, 0.05-1%의 농도범위에 걸쳐서 효과가 극대화되었다. 목향순기탕(木香順氣湯)과 강력한 항산화제인 DPPD는 $H_2O_2$에 의해 억제된 SOD의 활성에는 영향을 주지는 못했다. 그러나 $H_2O_2$에 의해 유발된 catalase, glutathione peroxidase, hydroperoxide 탈취효소의 활성이 감소되는 것을 억제하였다. 또한 $H_2O_2$에 의해 유발된 glutathione의 감소는 목향순기탕(木香順氣湯)과 DPPD에 의해 억제되었다. 목향순기탕(木香順氣湯)은 $H_2O_2$에 의해 유발된 ATP의 소실을 회복시켰지만 DPPD는 ATP 소실을 회복시키지 못하였다. 결론 : 이러한 결과로 볼 때 Caco-2세포에서 목향순기탕이 세포사망을 억제하는 것은 다른 기전을 통하여 항산화작용을 하는 것으로 볼 수 있다. 따라서 본 연구는 목향순기탕(木香順氣湯)이 반응성산소기에 의해 유발된 인체 위장관질환의 치료에 사용할 수 있을 가능성을 제시하고 있다.

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