• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1849-1855
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• 2015

Staphylococcus aureus plays an important role in sepsis, septic shock, pneumonia, and wound infections. Here, we demonstrate that Lactobacillus plantarum extracts inhibited S. aureus-induced cell death of a human epithelial cell line, HT-29. In particular, we have shown that S. aureus-induced cell death was abolished by neutralization of α-toxin, indicating that α-toxin is the major mediator of S. aureus-induced cell death. DNA fragmentation experiment and caspase assay revealed that the S. aureus-induced cell death was apoptosis. L. plantarum extracts inhibited the generation of effector caspase-3 and the initiator caspase-9 in S. aureus- or α-toxin-induced cell death. Moreover, expression of Bcl-2, an anti-apoptotic protein, was activated in L. plantarum extract-treated cells as compared with the S. aureus- or α-toxin-treated only cells. Furthermore, S. aureus-induced apoptosis was efficiently inhibited by lipoteichoic acid and peptidoglycan of L. plantarum. Together, our results suggest that L. plantarum extracts can inhibit the S. aureus-mediated apoptosis, which is associated with S. aureus spreading, in intestinal epithelial cells, and may provide a new therapeutic reagent to treat bacterial infections.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1253-1257
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• 2009

Exo-polysaccharide isolated from the culture of Grifola frondosa was modified by sodium periodate ($NaIO_4$) and sodium chlorite ($NaClO_2$) to delete polysaccharide part and phenolic compound, respectively, and was investigated what effect has each part of exo-polysaccharide against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in porcine kidney epithelial cells (LLC-PK1). Oxidative stress on LLC-PK1 cell was measured by cell viability, lipid peroxidation, superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) activity. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in significant decrease in cell viability, SOD, and GSH-px action, and significant increase in lipid peroxidation. The treatment of exo-polysaccharide and $NaIO_4$ modified sample protected LLC-PK1 cells from AAPH-induced cell damage such as cell viability, lipid peroxidation, SOD, and GSH-px activity in a dose dependant manner (10, 100, and $500{\mu}g/mL$). However, the treatment of $NaClO_2$ modified sample did not affect for cell viability, lipid peroxidation, SOD, and GSH-px activity. The antioxidant activity of exo-polysaccharide was significantly decreased on AAPH-induced LLC-PK1 cell system when phenolic compound was deleted. The antioxidant activity was significantly correlated with the content of phenolic compound of exo-polysaccharide.

• 대한의생명과학회지
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• 제4권2호
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• pp.77-86
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• 1998

백내장 환자의 수정체낭에서 연령차이에 따른 아교질과 type IV 아교섬유의 면역화학 반응 양상, apoptosis현상 및 수정체 상피세포의 전자현미경적인 변화를 관찰하여 다음과 같은 결과를 얻었다. Van Gieson염색소견은 수정체낭의 앞층과 앞아래층에서 짙은 염색반응이 있었으나 나이가 많을수록 반응이 감소되었다. Type IV 아교섬유 면역화학반응은 수정체낭의 앞층과 상피세포 주위에서 현저하게 일어났으며 나이가 많을수록 반응이 감소되었다. Periodic acid skiff-alcian blue반응의 변화는 수정체낭의 앞층에서 나이가 많을수록 현저하게 감소되었다. 세포자연사 현상은 수정체낭 상피세포의 핵내에서 일어났으며 나이가 많을 수록 반응이 일어나는 세포의 수가 감소되는 경향을 보였다. 수정체 상피세포의 전자현미경적인 변화는 40대까지는 외측 세포막의 folding (이하 주름)과 세포질내 공포가 증가되고 골지복합체가 발달되었으나 60대에서는 부분적으로 돌기는 내지만 골지복합체는 관찰되지 않았다. 수정체 상피세포의 바닥막부분에서는 나이가 들수록 거친 섬유들이 수정체 상피세포와 수정체낭을 격리시키고 있었으며, 수정체낭와 중간층은 40대군은 특징적으로 과립상의 구조물들이 집단적으로 축적된 부분들이 관찰되었고 60대군에서는 선상의 섬유성구조들이 나타났으며 짙은 전자밀도를 가진 과립의 수가 감소하였다.

• 동아시아식생활학회지
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• 제17권5호
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• pp.710-718
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• 2007

This study was designed to investigate the effects of plum(Prunus salicina Lindl. cultivars 'Oishiwase', 'Formosa', and 'Soldam') extracts on the proliferation as well as inhibition of human epithelial cells(HaCaT), human cervical carcinoma (HeLa, SiHa, and C33A) cells, and human stomach adenocarcinoma(SNU 638) cells. Dried plum was sequentially extracted and fractionated by hexane(KC-01), chloroform(KC-02), ethyl acetate(KC-03), n-butanol(KC-04), water(KC-05), methanol(KC-6), and hot water extract(KC-07). The epithelial and cancer cells were exposed for 48 h to $50{\mu}g/mL$ of plum extract in vitro, and were then analysed by a sulforhodamin B(SRB) staining assay. The methanol extract(KCP-6) of 'Formosa' proliferated not only the HaCaT cells(147.3%), but also the cervical carcinoma C33A cells(167.8%). The ethyl acetate extract of 'Soldam'(KCJ-3) significantly reduced the proliferation rate of the HPV positive conical carcinoma cells, at 61.5% for the SiHa cells and 70.5% for the HeLa cells. In the C33A cells, which are HPV negative cervical carcinoma cells, the hexane fractions of 'Formosa'(KCP-1) and 'Oishiwase'(KCD-1) markedly suppressed proliferation activity at 20.4% and 61.7%, respectively. However, the proliferation rate of the normal epithelial cells(HaCaT cell) was not reduced the proliferation rate by KCJ-3, KCP-1, or KCD-1, There were no significant effects on proliferation of the stomach cancer cells(SNU 638) by any of the extracts or fractions of the plum cultivars. These results suggest that the anti-proliferative effects of the plum cultivars were selective to the cancer cell origin. In conclusion, we found that several plum cultivar extracts, especially, the ethyl acetate fraction of 'Soldam" and the hexane fraction of "Formosa', have anti-proliferative activity toward human cervical carcinoma cells. However, further investigation is needed to assess the molecular mechanisms that mediate the antiproliferation activities of the plum cultivars.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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• pp.696-703
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• 2014

Clostridium difficile causes mucosal damage and diarrhea by releasing two exotoxins: toxin A and toxin B. C. difficile colitis is associated with alterations in bowel flora and the failure to mount an effective antibody response. The aim of the current study was to investigate whether antitoxin sera prevent toxin-A-induced apoptosis, cytoskeletal disaggregation, cell detachment, and tight junction loss in cultured colonic epithelial cells. Serum samples were isolated from mice that survived a C. difficile infection following antibiotic treatment, and the antitoxin effects of these samples were investigated in toxin-A-exposed HT29 colonic epithelial cells and a toxin-A-induced animal model of gut inflammation. Unchallenged mice did not produce IgG against toxin A, whereas serum (antiserum) from C. difficile-challenged mice showed significant IgG responses against toxin A. Treatment with the antiserum markedly inhibited mucosal damage and inflammation in the toxin-A-treated mouse model. In contrast to control mouse serum, the antiserum also markedly inhibited toxin-A-induced DNA fragmentation, dephosphorylation of paxillin and Epo receptor (EpoR), deacetylation of tubulin, and upregulation of p21(WAF1/CIP1) and p53. Taken together, these results reveal that the generated antitoxin serum has biotherapeutic effects in preventing various C. difficile toxin-A-induced cellular toxicities.

• The Korean Journal of Physiology and Pharmacology
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• 제16권5호
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• pp.313-320
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• 2012

In this study, we focused to identify whether eupatilin (5,7-dihydroxy-3',4',6-trimethoxyflavone), an extract from Artemisia argyi folium, prevents $H_2O_2$-induced injury of cultured feline esophageal epithelial cells. Cell viability was measured by the conventional MTT reduction assay. Western blot analysis was performed to investigate the expression of 5-lipoxygenase by $H_2O_2$ treatment in the absence and presence of inhibitors. When cells were exposed to 600 ${\mu}M$ $H_2O_2$ for 24 hours, cell viability was decreased to 40%. However, when cells were pretreated with 25~150 ${\mu}M$ eupatilin for 12 hours, viability was significantly restored in a concentration-dependent manner. $H_2O_2$-treated cells were shown to express 5-lipoxygenase, whereas the cells pretreated with eupatilin exhibited reduction in the expression of 5-lipoxygenase. The $H_2O_2$-induced increase of 5-lipoxygenase expression was prevented by SB202190, SP600125, or NAC. We further demonstrated that the level of leukotriene $B_4$ ($LTB_4$) was also reduced by eupatilin, SB202190, SP600125, NAC, or nordihydroguaiaretic acid (a lipoxygenase inhibitor) pretreatment. $H_2O_2$ induced the activation of p38MAPK and JNK, this activation was inhibited by eupatilin. These results indicate that eupatilin may reduce $H_2O_2$-induced cytotoxicity, and 5-lipoxygenase expression and $LTB_4$ production by controlling the p38 MAPK and JNK signaling pathways through antioxidative action in feline esophageal epithelial cells.

• 대한기관식도과학회:학술대회논문집
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• 대한기관식도과학회 1993년도 제27차 학술대회 초록집
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• pp.98-98
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• 1993

편도선의 염증반응은 세균이 편도상피세포에 부착되는 것으로부터 시작되며 부착된 세균은 증식하여 집락을 형성한 후 독소를 분비하여 점막 장벽을 뚫고 조직에 손상을 일으킨다. 저자들은 급성편도선염의 병태를 이해하기 위하여 생체내에서 급성편도선염 환자군과 정상인에서 상피세포의 세균부착성에 대한 차이를 알아보고자 본 실험을 시행하였다. 급욍편도선염 환자군 20례와 정상인 대조군 20례를 대상으로 편도선부위를 면봉으로 문지른후 세포혼합물을 Acridine orange로 염색하여 형광현미경하에서 관찰하였다. 50개의 편도상피세포에서 부착된 세균수를 계산하였다. 또한 동시에 근배양을 시행하였다. 급성편도선염군은 대조군보다 상피세포에서 10% 이상 부착된 세균수가 많았으며 (p<0.05), 상피세포에 부착된 세균수는 연령과 유의한 상관관계를 보여주었다.

• Archives of Pharmacal Research
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• 제21권4호
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• pp.398-405
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• 1998

We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. $G_2$/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of $G_1$ phase cells was not observed. The colonies with distinct multicelfular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.

• Journal of Pharmaceutical Investigation
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• 제32권1호
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• pp.21-26
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• 2002

The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

• 대한본초학회지
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• 제38권5호
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• pp.1-7
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• 2023

Purpose : To compare the skin damage recovery efficacy of natural Angelicae Gigantis Radix extract (N-AGR) and cultivated A. Gigantis Radix extract (C-AGR). Through this, we confirmed whether the quality standards of herbal medicines recorded in the classic books make a difference in the experimental efficacy using epithelial cells. Methods : The quality standards of medicinal herbs in the classic books and the cultivation and processing conditions of two types of A. gigas were compared. After inducing oxidative stress with H₂O₂, cytoprotective property of N-AGR and C-AGR were evaluated through cell viability. Additionally, after wound formation of epithelial JB6 cells, N-AGR and C-AGR were treated to evaluate wound healing efficacy. Result : The natural A, gigas met the excellent quality standards of the classic books. N-AGR inhibited cell death by oxidative stress induced by H₂O₂, and was superior to C-AGR. Both N-AGR and C-AGR showed dose-dependent wound healing efficacy, but N-AGR was significantly superior to C-AGR. Conclusions : Through the oxidative stress of skin and skin wound healing efficacy experiments using epithelial cells, natural A. gigas showed superior efficacy compared to cultivated A. gigas.