• 한국식품과학회지
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• 제48권4호
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• pp.306-316
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• 2016

멸치 젓갈로부터 분리한 젖산세균을 대상으로 프로바이오틱 균주로서의 기능적 특성과 안전성을 평가하였다. 분리된 균주는 생화학적 특성과 당 발효능 및 염기순서 분석을 통해 Enterococcus faecium AJ06, Leuconostoc mesenteroides AJ13, Pediococcus halophilus AJ22, Lactobacillus sakei AJ29 및 Pediococcus pentosaceus AJ35로 동정되었다. AJ06, AJ22 및 AJ29 균주들은 펩신이 첨가된 인공 위액 및 쓸개즙액에서 강한 저항성을 나타내었고, taurocholic acid 혹은 taurodeoxycholic acid가 첨가된 MRS 한천 평판 배지 상에서 쓸개즙분해효소를 생산하였다. 특히, Caco-2 세포에 대한 높은 부착능과 다양한 항생제에 대한 저항성을 나타낸 AJ22와 AJ29 균주는 항균물질 생산으로 인해 식중독균의 증식을 효과적으로 저해하였다. 이들 균주는 혈액 한천 평판배지 상에서 알파 및 베타용혈독을 나타내지 않았고, 아미노산 전구체가 함유된 MRS 액상배지 내에서 생체 아민을 생산하지 않았으며, Salmonella Typhimurium TA98 and TA100 균주에 대한 돌연변이도 유발하지 않았다.

• BMB Reports
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• 제46권1호
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• pp.47-52
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• 2013

Intestinal epithelial cell (IEC) apoptosis induced by hypoxia compromise intestinal epithelium barrier function. Both Akt and Hsp90 have cytoprotective function. However, the specific role of Akt and $Hsp90{\beta}$ in IEC apoptosis induced by hypoxia has not been explored. We confirmed that hypoxia-induced apoptosis was reduced by $Hsp90{\beta}$ overexpression but enhanced by decreasing $Hsp90{\beta}$ expression. $Hsp90{\beta}$ overexpression enhanced BAD phosphorylation and thus reduced mitochondrial release of cytochrome C. Reducing $Hsp90{\beta}$ expression had opposite effects. The protective effect of $Hsp90{\beta}$ against apoptosis was negated by LY294002, an Akt inhibitor. Further study showed that Akt phosphorylation was enhanced by $Hsp90{\beta}$, which was not due to the activation of upstream PI3K and PDK1 but because of stabilization of pAkt via direct interaction between $Hsp90{\beta}$ and pAkt. These results demonstrate that $Hsp90{\beta}$ may play a significant role in protecting IECs from hypoxia-induced apoptosis via stabilizing pAkt to phosphorylate BAD and reduce cytochrome C release.

• 대한한의학회지
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• 제23권3호
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• pp.164-173
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• 2002

목적：반응성산소기들은 장관에서 여러 종류의 질병의 발생과 관련을 가지고 있는 것으로 알려져 있어, 이들에 의한 세포손상을 방지하는 약물의 개발은 시급한 실정이다. 본 연구에서는 항산화작용을 가진 약재로 보고 된 단삼추출액이 장관상피세포에서 $H_2O_2$에 의한 세포손상을 방지할 수 있는 지를 조사하고자 하였다. 방법：장관상피세포로는 사람의 소장상피세포에서 유래한 배양세포주인 Caco-2세포를 이용하였고, 세포손상 정도는 trypan blue exclusion assay를 통해 평가하였고, 지질의 과산화는 그 산물인 malondialdehyde의 량을 측정하여 산정하였다. 결과： $H_2O_2$는 처리 시간 및 농도에 비례하여 세포손상을 유발하였으며, 이러한 효과는 단삼추출액에 의해 농도의존적으로 방지되었다. $H_2O_2$에 의한 세포소상은 $H_2O_2$제거제인 catalase와 철착염제인 deferoxamine에 의해 방지되었으나 항산화제인 N,N-diphenyl-p-phenylenamine(DPPD)에 의해 영향을 받지 않았다. $H_2O_2$는 지질의 과산화를 증가시켰으며, 이러한 효과는 단삼추출액과 DPPD에 의해 억제되었다. 단삼추출액은 $H_2O_2$에 의한 세포내 ATP 고갈을 방지하였다. $H_2O_2$는 DNA 손상을 일으켰으며, 이러한 효과는 단삼추출액, catalase 및 deferoxamine에 의해 방지되었으나, DPPD에 의해서는 변화되지 않았다. 결론 : 이상의 결과를 종합하면 단삼추출액은 장관상피세포에서 $H_2O_2$에 의한 세포손상을 방지하며, 이러한 효과는 항산화작용이 아닌 다른 작용기전에 기인할 것으로 생각된다. 또한 본 연구의 결과는 $H_2O_2$가 장관상피세포에서 지질의 과산화를 유발하여 세포손상을 일으키지 않음을 가리킨다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.743-748
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• 2016

Background: $K^-Ras$ activation is an early event in colorectal carcinogenesis and associated mutations have been reported in about 40% of colorectal cancer patients. These mutations have always been responsible for enhancing malignancy and silencing them is associated with attenuation of tumorigenicity. Among downstream effectors are the RAF/MEK/ERK and the PI3K/Akt signaling pathways. PI3K/Akt signaling leads to reduction of apoptosis, stimulated cell growth and enhanced proliferation. Ellagic acid (EA), a naturally occurring antioxidant, has recently emerged as a promising anti-cancer agent. Purpose: To evaluate the impact of cellular genetic makeup of two colon cancer cell lines with different genetic backgrounds, HCT-116 ($K^-Ras^-/p53^+$) and Caco-2 ($K^-Ras^+/p53^-$), on response to potential anti-tumour effects of EA. In addition, the influence of $K^-Ras$ silencing in HCT-116 cells was investigated. Materials and Methods: Cellular proliferation, morphology and cell cycle analysis were carried out in addition to Western blotting for detecting total Akt and p-Akt (at Thr308 and Ser473) in the presence and absence of different concentrations of EA. Cell proliferation was also assessed in cells transfected with different concentrations of $K^-Ras$ siRNA or incubated with ellagic acid following transfection. Results: The results of the present study revealed that EA exerts anti-proliferative and dose-dependent pro-apoptotic effects. Cytostatic and cytotoxic effects were also observed. p-Akt (at Thr308 and Ser473) was downregulated. Moreover, EA treatment was found to (i) reduce $K^-Ras$ protein expression; (ii) in cells transfected with siRNA and co-treated with EA, pronounced anti-proliferative effects as well as depletion of p-Akt (at Thr308) were detected. Conclusions: Cellular genetic makeup ($K^-Ras^-/p53^-$) was not likely to impose limitations on targeting EA in treatment of colon cancer. EA had a multi-disciplinary pro-apoptotic anti-proliferative approach, having inhibited Akt phosphorylation, induced cell cycle arrest and showed an anti-proliferative potential in HCT-116 cells (expressing mutant $K^-Ras$).

• ALGAE
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• 제35권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.225-235
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• 2014

Total 121 lactic acid bacteria were isolated from homemade Inner Mongolia extra hard Hurood cheese. Seven of these strains, identified as Lactobacillus plantarum, were studied for probiotic characteristics. All seven strains survived at pH 3.0 for 3 h, or in the presence of oxgall at 0.3% or 0.6% for 4 h, but their viabilities were affected to different extents at pH 2.0 for 3 h. Strains C37 and C51 showed better adherence to Caco-2 cells, and higher hydrophobicity. The seven L. plantarum strains were different in in vitro free radical scavenging activities and cholesterol-reducing ability. In vivo evaluation of the influence of L. plantarum C37 on the intestinal flora in a mouse model showed strain C37 could increase the viable counts of lactobacilli in feces of mice and decrease the viable counts of enterococci. When L. plantarum C37 was used to prepare probiotic Hurood cheese, it was able to maintain high viable counts (>7.8 log CFU/g) during the whole storage period, but the composition of the cheese was not changed. These results indicate that L. plantarum C37 could be considered as a promising probiotic strain.

• Archives of Pharmacal Research
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• 제26권9호
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• pp.768-772
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• 2003

Tween 80 (Polysorbate 80) is a hydrophilic nonionic surfactant commonly used as an ingredient in dosing vehicles for pre-clinical in vivo studies (e.g., pharmacokinetic studies, etc.). Tween 80 increased apical to basolateral permeability of digoxin in Caco-2 cells suggesting that Tween 80 is an in vitro inhibitor of P-gp. The overall objective of the present study was to investigate whether an inhibition of P-gp by Tween 80 can potentially influence in vivo absorption of P-gp substrates by evaluating the effect of Tween 80 on the disposition of digoxin (a model P-gp substrate with minimum metabolism) after oral administration in rats. Rats were dosed orally with digoxin (0.2 mg/kg) formulated in ethanol (40％, v/v) and saline mixture with and without Tween 80 (1 or 10％, v/v). Digoxin oral AUC increased 30 and 61％ when dosed in 1 ％ and 10％ Tween 80, respectively, compared to control (P＜0.05). To further examine whether the increase in digoxin AUC after oral administration of Tween 80 is due, in part, to a systemic inhibition of digoxin excretion in addition to an inhibition of P-gp in the GI tract, a separate group of rats received digoxin intravenously (0.2 mg/kg) and Tween 80 (10％ v/v) orally. No significant changes in digoxin IV AUC was noted when Tween 80 was administered orally. In conclusion, Tween 80 significantly increased digoxin AUC and Cmax after oral administration, and the increased AUC is likely to be due to an inhibition of P-gp in the gut (i.e., improved absorption). Therefore, Tween 80 is likely to improve systemic exposure of P-gp substrates after oral administration. Comparing AUC after oral administration with and without Tween 80 may be a viable strategy in evaluating whether oral absorption of P-gp substrates is potentially limited by P-gp in the gut.

• 한국식품과학회지
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• 제50권4호
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• pp.414-419
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• 2018

병원성 대장균 O157:H7은 식중독 사고를 일으키는 주된 원인균으로서 항생제 내성 문제를 극복하기 위해서는 병원성 대장균 O157:H7을 제어하기 위한 새로운 방법이 개발되어야 한다. 따라서 본 연구에서는 식물화학물질을 스크리닝하여 병원성 대장균 O157:H7의 부착에 주된 역할을 하는 LEE 오페론의 발현을 감소시키는 물질을 찾고자 하였다. 스크리닝을 통해 선발된 식물화학물질 중 다이오스민은 사람 결장 상피세포 부착능을 3.62배 감소시킴으로써(p<0.01) 양성대조군으로 사용된 미리세틴과 유사한 정도의 효과를 나타냈다. 생물막 형성에 있어서는 다이오스민 처리 시 표현형이 25.6% 감소하여 유의적 차이가 확인되었고(p<0.05), curli 유전자의 발현 역시 미리세틴 처리 때보다 1.57-2.60배 더 유의미하게 감소하는 것으로 나타나 양성대조군보다 더 좋은 효과를 보였다. 한편 다이오스민은 병원성 대장균 O157:H7의 생장에는 아무런 영향을 미치지 않는 것으로 나타나 내성 발생률이 저감화된 새로운 항미생물제재로서 사용 가능하리라 판단된다.

• Journal of Microbiology and Biotechnology
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• 제26권10호
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• pp.1675-1686
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• 2016

We characterized the probiotic properties of Lactobacillus helveticus strains KII13 and KHI1 isolated from fermented cow milk by in vitro and in vivo studies. The strains exhibited tolerance to simulated orogastrointestinal condition, adherence to Caco-2 cells, and antimicrobial activity. Both L. helveticus strains produced bioactive tripeptides, isoleucylprolyl-proline and valyl-prolyl-proline, during fermentation of milk. KII13 showed higher in vitro cholesterol-lowering activity (47%) compared with KHI1 (28%) and L. helveticus ATCC 15009 (22%), and hence, it was selected for in vivo study of cholesterol-lowering activity in atherogenic diet-fed hypercholesterolemic mice. For the study, mice were divided into four groups (viz., normal diet control group, atherogenic diet control group (HCD), KII13-atherogenic diet group (HCD-KII13), and Lactobacillus acidophilus ATCC 43121-atherogenic diet group (HCD-L.ac) as positive control). The serum total cholesterol level was significantly decreased by 8.6% and 7.78% in the HCD-KII13 and HCD-L.ac groups (p < 0.05), respectively, compared with the HCD group. Low-density lipoprotein cholesterol levels in both HCD-KII13 and HCD-L.ac groups were decreased by 13% and 11%, respectively, compared with the HCD group (both, p < 0.05). Analysis of cholesterol metabolism-related gene expression in mice liver showed increased expression of LDLR and SREBF2 genes in mice fed with KII13. By comparing all the results, we conclude that L. helveticus KII13 could be used as a potential probiotic strain to produce antihypertensive peptides and reduce serum cholesterol.

• 생약학회지
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• 제53권4호
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• pp.213-225
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• 2022

Schisandra chinensis is widely known to regulate fatigue recovery, lowering blood pressure, and contains several bioactive compounds such as schizadran. In this study, we obtained basic data for the development of health food by measuring the schizandrin content, antioxidant activity, anti-inflammatory, antibacterial activity, and enzyme activity of the hot water extract (SCW) and 40% ethanol extract (SCE) of S. chinensis grown in Sunchang-gun, Korea. Schizandrin content was measured as 7.87 ± 0.01 mg/g for SCW and 10.38 ± 0.05 mg/g for SCE. SCE had higher DPPH and ABTS radical scavenging activity than SCW, and the total polyphenol and flavonoid contents were also higher. Both SCW and SCE (500 ㎍/mL) exhibited more than 55% protective effects against oxidative stress in HepG2 cell lines. Anti-inflammatory efficacy was assessed using RAW 264.7 and Caco-2 cells and both SCW and SCE do not have cytotoxic effects. Anti-inflammatory results showed concentration-dependent NO activity and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) inhibition. The antibacterial activity increased with increasing dose concentrations of SCW and SCE, and the MIC was 25 mg/mL for L. monocytogenes, S. typhimurium, and 75 mg/mL for H. pylori. In addition, amylase and protease enzyme activity was observed in both SCW and SCE.