Helicobacter pylori are known as a causative agent of gastritis, gastric duodenum and peptic ulcer, and gastric cancer, and multiple drug use is associated with various side effects in patients. The discovery of antibacterial substances against H. pylori from Korean resource plants is an important substitute for antibiotics. 52 species of Korean resource plants were collected and extracted with 50% ethanol, and antibacterial activity against H. pylori was measured using the disk diffusion method. The toxicity of plant extracts to human gastric adenocarcinoma(AGS) cells was measured by MTT assay, and the level of IL-8 secreted when gastric epithelial cells were inoculated with H. pylori was measured. As a result of measuring the antibacterial activity of H. pylori, antibacterial activity was confirmed in 38 plant extracts. The plant species with the strongest antibacterial activity were Chrysanthemum indicum, Rheum rhabarbarum, Patrinia scabiosaefolia and Petasites japonicus. C. indicum was not cytotoxic to H. pyroli-infected AGS cells and showed anti-inflammatory effects. This study's results can be used to develop healthy, functional foods and medical materials.

Helicobacter (H.) pylori is one of the most common pathogenic bacteria in the human gut, and is also one of the most important factors that cause digestive disorders such as chronic inflammation, gastric ulcers, and even gastric cancer. Since the use of various antibiotics to treat H. pylori infection is associated with the development of resistance in this bacterium, the aim of this study was to determine the anti-H. pylori effects of Peucedanum japonicum extracts (PJE) in the stomach tissue of C57BL/6 mice. In this experimental study, 50 C57BL/6 mice in five groups were evaluated. After induction of H. pylori infection in mice with the standard strain of H. pylori (ATCC 43504), the infected mice were treated with 3 different doses of PJE in different groups. Then, the anti-H. pylori effects of PJE were evaluated by rapid urease test (CLO), histopathological exam and blood interleukin (IL)-8 ELISA analysis. Based on CLO results, H. pylori eradication effects of PJE was observed. H. pylori infection of C57BL/6 mice showed significant increase of histopathological lesion scores. However, the treatment of PJE showed significant decrease of histopathological lesion scores. The treatment of PJE decreased gastric inflammation and mucosal lesions in the stomach of H. pylori-infected mice. Blood IL-8 levels were elevated 4 weeks after H. pylori inoculation. However, the treatment of PJE showed significant decrease of IL-8 level. According to the results of this study, PJE could be useful in the reduction of H. pylori infection in the mouse model.

Dental health care workers (DHCW) are at a risk of occupational exposure to Helicobacter pylori from the aerosolized oral biofilms and saliva of patients. We designed this study to investigate the prevalence of H. pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka. After obtaining informed consent, oral biofilms were collected from 38 dental undergraduates (19 males and 19 females) undergoing clinical training and 33 non-dental undergraduates (14 males and 19 females). The participants were in the age range of 22–27 years and had healthy periodontium. Total DNA from the oral biofilms were extracted, and H. pylori DNA was detected using polymerase chain reaction (PCR) amplification of 16S rRNA gene of H. pylori using JW22-JW23 primers, and the results were confirmed using PCR amplification of H. pylori -urease specific HPU1-HPU2 primers. Out of 71 participants, 11 (28.95%) dental and 3 (9.09%) non-dental undergraduates had H. pylori in their oral biofilms indicating an overall prevalence rate of 19.72% (14/71). Thus, the prevalence of H. pylori in oral biofilms was significantly higher in dental undergraduates than in non-dental undergraduates (p < 0.05). An odds ratio of 4.07 indicated that dental undergraduates were four times more likely to harbor H. pylori in their oral biofilms than nondental undergraduates. Foregoing data support the fact that there may be greater occupational risk of exposure to H. pylori for dental undergraduates during clinical training than that for non-dental undergraduates, warranting meticulous infection control practices during clinical dentistry.

The efficacy of standard therapeutic strategies for Helicobacter pylori infection is decreasing over time due to the emergence of drug-resistant strains. At the our previous study, we reported that the lactic acid bacterium Lactobacillus paracasei HP7 isolated from kimchi - a fermented vegetable dish widely consumed in Korea - had inhibitory effects against H. pylori in vitro and in vivo. In this study, we are aimed to investigate the synergistic effect of probiotic HP7 and the extracts of Perilla frutescens (PFE) and Glycyrrhiza glabra (GGE) to alleviate Helicobacter pylori-induced damages. H. pylori infection in KATO III cells decreased mucin gene expression over time. The reduction of mucin gene expression was significantly recovered at 6 hours when GGE, PFE, and Lactobacillus HP7 were co-treated. H. pylori infection of C57BL/6 mice was demonstrated by H. pylori-IgG concentration, and results showed significant increase in mice post-infected with H. pylori. Blood Tumor Necrosis Factor (TNF)-α levels were elevated 4 weeks after H. pylori inoculation. However, the treatment of a combined mixture of HP7 including GGE and PFE induced significant decrease of TNF-α level (p<0.01) and showed significant decrease of TNF-α level as compared as the level by the positive control Glycyrrhiza glabra acetone extract (Deglycyrrhizinated Licorice, DGL) treatment (p<0.05). Additionally, the treatment of a combined mixture of HP7 including GGE and PFE decreased gastric inflammation and mucosal lesions in the stomach of H. pylori-infected mice. These results demonstrate that he treatment of a combined mixture of HP7 including GGE and PFE treatment can inhibit H. pylori growth and is thus a promising treatment for patients with gastric symptoms such as gastritis that are caused by H. pylori infection.

만성 위염, 위암, 위궤양 등을 유발하는 Helicobacter pylori는 항생제의 부작용, 내성, 다제 내성균의 등장으로 항생제만으로 안전하게 치료하기는 어려운 실정이다. 안전하게 제균을 하기 위해 항생제를 대체할 수 있는 천연물 소재 치료제를 개발하기 위해 많은 연구들을 하고 있다. 선행 연구를 통해 in vitro 실험 H. pylori 항균 효과 평가를 통해 항균 효과가 우수한 추출물을 알아보고자 한다.

본 실험은 위점막에 존재하는 H. pylori에 대한 PJT 추출물의 생장 억제를 알아보고자 하였다. H. pylori에 대한 치료제로는 항생제가 있으나 높은 내성률 등의 문제가 야기되 고 있다. 천연물은 화합물보다 높은 안전성과 항생제 내성 문제의 해결 측면에서 큰 장점을 가지고 있어 예방 및 치료제의 개발로 많은 연구가 진행되고 있다. PJT는 활성화와 항알레르기 작용 및 실험성 위궤양의 항궤양 작용에 우수한 것으로 알려져 있다. 또한 뇌 졸중으로 발병되는 중풍병, 해독 등의 효능이 있어 활용성이 높은 약초로 향후 연구대상 약용작물로 각광을 받을 가능성이 큰 항균성 약용작물이다. 실험에서 사용된 H. pylori 균은 멸균된 플레이트에 접종한 후, 10% CO2 및 100% 습 도가 유지되는 37℃ incubation에 3일간 배양 후 평가하였다. 균이 접종된 플레이트에 각각 표준자를 이용하여 6시간, 12시간, 24시간 및 48시간 후 clear zone의 지름으로 표현되는 억제범위(the zone of inhibition)를 측정하여 균이 자라지 않은 clear zone을 관찰하였다. 실험 종료 후, H. pylori 생장 억제 활성 측정결과 에탄올, 열수 추출물에서의 생장 억제를 확인할 수 있었다. 본 실험에서 열수 추출보다 에탄올 추출에서 더 나은 저해 활성을 나타난 것을 확인할 수 있었고, 이것은 물과 에탄올 즉 용매의 극성이 각각 다르기 때문에 차이가 나는 것으로 사료된다. 따라서 PJT은 H. pylori를 효과적으로 억제하는 것으로 판단되며, 향후 H. pylori에 대한 갯기름나물 항궤양 효과에 지표로 쓰일 수 있을 것으로 판단된다.

Infection with Helicobacter pylori leads to gastritis, peptic ulcers and gastric cancer. Moreover, when the gastric mucosa is exposed to H. pylori, gastric mucosal inflammatory cytokine interleukin-8 (IL-8) and reactive oxygen (ROS) species increase. Tribulus terrestris L. is an annual creeping herb of the family Zygophyllaceae. In this study, the effect of Tribulus terrestris L. fruits extract (TTE) extract on H. pylori-infected human gastric epithelial cells was examined. Cell viability was determined by the tetrazolium salt reduction method using WST-1 according to the manufacturer's instructions. AGS cells were pretreated with TTE extract for 24 hrs followed by H. pylori infection for up to 24 hrs. IL-8 secretion in AGS cells was measured by ELISA. The extract yield of the fruits of Tribulus terrestris with 50% ethanol was 20.60%. We analyzed TFE composition by LC. The concentration of Protodioscin in TFE was 310 μg/mL. It was confirmed that exposure to 100 μg/mL of TTE had no significant effect on cell proliferation at the concentrations examined (12.5–200 μg/mL). It was therefore concluded that TTE at these concentrations had no cytotoxic effects on AGS cells and could be used in this study. Pretreatment of H. pylori-infected AGS cells with 12.5, 25, 50 and 100 μg/mL of TTE for 24 hrs significantly decreased IL-8 production by 12.5%, 25%, 27.5% and 50%, respectively, compared to H. pylori-infected cells without TTE. In this study, we found that TTE inhibited H. pylori-induced IL-8 secretion, thus augmenting their benefit in regard to protection of gastric epithelial cells. This study suggests that ingestion of these plant extracts could have therapeutic implications for patients with H. pylori induced gastritis and duodenal ulcer.

Propolis possesses antimicrobial, anti-inflammatory, anaesthetic and immunostimulating activities. The aim of this study was to evaluated chemical composition and in vitro anti-Helicobacter pylori activity of eight propolis ethanolic extracts form different region in Korea. The chemical composition of eight Korean propolis was investicated by absorption spectrophotomety and UHPLC. Total penolic and total flavonoids content ranged between 21±1.5 and 31±1.8 gallic acid g/100 g and 7.2±0.9 and 24.4±1.9 quercetin g/100 g, respectively. The p-coumaric acid as standard components content ranged between 0.29±0.00 and 2.34±0.02 mg/g using UHPLC. Cinnamic acid content between 0.43±0.00 and 1.33±0.01 mg/g. 80% ethanolic extract of propolis against H. pylori was evaluated by using the disc diffusion method. Ethanol was used as a control. Mean diameters of H. pylori growth inhibition using 10 μg/mL, 50 μg/mL or 100 μg/mL ethanol propolis per disc were 0, ranging from 0.9 to 2.1 mm and from 1.9 to 3.5 mm, respectively. In conclusion, variation in the chemical composition of Korean propolis were mainly due to the difference in the preferred regional plants visited by honeybees. Also Korean propolis possesses considerable antibacterial activity against H. pylori. There was not confirmed a correlation between the chemical composion contens and anti-H. pylori effect. The potential of propolis in the prevention or treatment of H. pylori infection is worth further extensive evaluation.

비피도박테리아 증식인자인 1,4-dihydroxy-2-naphthoica cid 생산 유산균주 LP2가 선행연구에서 자연치즈로부터 분리되었다. LP2 균주는16S rRNA 유전자 염기서열 분석에 의한 균주동정에서 Lactobacillus plantarum(99% 일치율)과 가장 높은 상동성을 나타내어 Lactobacillus plantarum LP2 균주로 명명하였다. LP2 균주의 배양 상등액은 Helicobacter pylori KCTC 12083 균주에 대하여 항균활성을 나타내었으며 이는 생산된 DHNA에 기인하는 것으로 추측되었다.

Vitamin C (ascorbic acid) is an essential nutrient of most living tissues. We established a strain of Gulo-/- mice with known deficiency, in which vitamin C intake can be controlled by diet, like humans, and investigated the differen- tially expressed proteins following treatments with Helicobacter pylori and diethylnitrosamine (DENA) in the liver of Gulo-/- mice using a proteomic approach. Expression of p53, 14-3-3ε and 14-3-3δ in Gulo-/- mice liver tissue was analyzed by immunohistochemistry. 2-DE maps constructed from Gulo-/- mice liver and differentially expressed proteins in liver tissue were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF/MS). In Gulo- /- mice after H. Pylori infection, followed by treatment with DENA, no differences in p53, 14-3-3ε and 14-3-3δ were observed by immunohistochemistry. Proteome analyses us- ing MALDI-TOF/MS resulted in successful identification of 12 proteins (nine proteins were up-regulated and three were down-regulated). Specifically, peroxiredoxin-6 and Alpha-1-antitrypsin 1-4 were up-regulated in liver after H. Pylori infection followed by treatment with DENA. These results indicated that oral supplementation with vitamin C led to rescue of Gulo-/- mice from vitamin deficiency, and protected the liver from H.pylori infection and/or DENA ef- fect, and vitamin C also protected the liver against oxidative stress.

소화성 질환의 중요 인자인 Helicobacter pylori를 저해하는 IgY, 목이버섯, 김치와 타락 유산균을 이용하여 생육 저해 상승 효과를 분석하고자 하였다. 동결 건조된 유산균 배양 상징액을 다양한 효소 처리 결과, 지질 분해 효소를 제외하고는 활성을 나타냈다. GC 분석을 통해 유산균 동결건조액의 지방산 조성은 undecanoic acid(C11:0), palmitic acid(C16:0), steraic acid(C18:0), oleic acid(C18:1)가 L. mensenteroides LAB KW5와 S. thermophilus LAB KW15에서 모두 확인되었으며, eicosadienoic acid(C20:2)는 LAB KW5에서만 나타났다. 또한 유산균 동결건조액은 다른 식중독균에서도 spot assay의 결과, 그람 음성균 중에서 특히 E. coli O157:H7, E. coli, C. sakazakii 등에서 생육 저해능이 확인되었다. 목이버섯 추출물은 열수 추출과 ethanol를 이용해 분리하였으며, HPLC를 이용하여 목이버섯 추출 다당체를 분석한 결과, glucuronoxylomannan 혹은 glucomannan이 β-glucan과 함께 존재하는 혼합물일 것으로 되었다. 또한, 면역란은 1차 접종 후 11일째부터 주마다 2회씩 회수하여 IgY를 분리, 정제하였다. 실험을 통해 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액을 혼합 배양하여 배양시간에 따른 생육 저해력의 변화를 확인한 결과, 유산균에 의해 H. pylori의 저해 효과가 있었으며, IgY와 목이버섯의 혼합 시 추가 억제 효과가 있는 것으로 나타났다. 그러므로 동결 건조된 유산균 배양 상징액, IgY, 목이버섯 추출액의 복합처리는 H. pylori를 제어하는데 효과적인 것으로 보인다.

In this study, a medicinal herbal plant, Cinnamomum cassia, was extracted by three different methods using water, methanol, or ethanol. For anti-helicobacter activity screening, inhibitory zone tests as an in vitro assay were performed respectively with the extracting compounds. As the result of inhibitory zone test, Cinnamomum cassia extract exhibited strong anti-helicobacter activity. In addition, we performed a comparison of the antibacterial activities according to the extracting methods of Cinnamomum cassia against Helicobacter pylori. The 70% ethanol-extracted compound exhibited stronger anti-helicobacter activity than the compounds extracted with water or methanol. These results indicate that it can be used for treatment against H. pylori infection and protected against H. pylori-induced pathology using 70% ethanol-extracted Cinnamomum cassia.

It has been suggested that Helicobacter pylori(H. pylori) infections can promote the development and progression of gastric cancer through the modulation of cell cycle regulators such as p27Kip1 and Skp2. p27Kip1 is a cyclin-dependent kinase (CDK) inhibitor that blocks the G1/S transition necessary for cell cycle progression. Skp2 is a component of the ubiquitin ligase complex called SCFSkp2(SKP1-Cullin-F-box), which specifically binds and promotes the degradation of p27Kip1. A low level of p27Kip1 and a high level of Skp2 have been reported in many types of cancers, including gastric cancer. In addition, a decrease in p27Kip1 has been reported in H. pylori-infected specimens. However, data on Skp2 in H. pylori infections are limited. This study examines the changes in the status of Skp2 in H. pylori-infected gastric epithelial AGS cells. For this, we stimulated AGS cells with H. pylori(NCTC 11637) at the ratio of 300:1(bacterium:cell) for 6 hours. The results of an immunoprecipitation analysis, followed by a western blot, indicate that the interaction between Skp2 and 14-3-3 was elevated 3 hours after the H. pylori treatment. In addition, there was an increase in cytoplasmic Skp2 after 3 hours, whereas there was no change in the nuclear level. Since it has been reported that interaction with 14-3-3 and the subsequent cytoplasmic translocation of Skp2 can increase its protein stability, increases in the interaction with 14-3-3 and the cytoplasmic Skp2 after the H. pylori treatment can increase the level of Skp2 in AGS cells. This phenomenon may explain, at least to some extent, the mechanism underlying the relationship between H. pylori infections and gastric carcinogenesis.

Yoghurt는 쉽게 접할 수 있는 기호성 식품으로 생체 조절 기능을 갖는 기능성과 다양한 맛을 부여할 수 있는 퓨전성을 동시에 강화시킬 수 있기 때문에 여러 가지 기능성 소재들을 첨가하여 새로운 yoghurt를 개발하기 위한 연구가 활발하게 수행되고 있다. 본 연구는 probiotics 유산균이 생성하는 천연의 세포 외 다당류(EPS; exopolysaccharide)를 이용하여 품질이 개선된 Helicobacter pylori을 억제하는 기능성 yoghurt를 제조하고자 우리나라의 식품공전에 식물성 재료로 허가되어 있어 사용 시 안전성에 문제가 없으며 전래된 여러 효능에 착안하여 감초를 이용한 기능성 yoghurt를 개발을 검토하였다. 감초 용매 추출물(1 mg/mL)의 H. pylori KCCM 40449에 대한 항균 활성을 측정한 결과, 40% methanol 추출물은 13.2±0.6 mm, 40% ethanol 추출물은 14.0±0.2 mm, 80% ethanol 추출물은 14.3±0.3 mm, 40% isopropanol 추출물은 15.1±0.3 mm의 생육저지환을 나타내었으며(p < 0.05), 최소 저해 농도(Minimum Inhibitory concentration: MIC)는 각 용매 추출물의 경우 25-100 μg/mL로 나타났다. 40% 에탄올 감초 추출물을 우유 배지에 0.05, 0.1, 0.2%(v/v)의 농도로 첨가하고 Lactobacillus amylovorus DU-21을 접종하여 37oC에서 72시간 동안 발효시키면서, yoghurt의 이화학적 특성 및 관능적 특성을 관찰하였다. pH는 균 접종 직후, 6.44-6.47에서 72시간 발효 후, 3.41-3.51를 나타내었으며, 적정산도는 0.109-0.111%에서 1.021-1.091%로 변화하였다. 점도와 syneresis를 측정한 결과, yoghurt의 점도는 발효 12시간까지 모든 실험구가 급격히 증가하였으며, 그 이후 24시간까지 증가 폭이 다소 둔화된 후, 72시간까지 완만한 증가 양상을 나타내었다. Syneresis는 발효 시간이 증가할수록 감소하였다. 72시간 발효 하였을 때, 감초추출물 0.2% 첨가구에서 12.0±0.4%로 가장 적은 값을 나타내었다. 산생성이 진행됨에 따라 점도가 증가하였으며 syneresis가 감소하는 것을 알 수 있었다. Yoghurt의 Helicobacter pylori KCCM 40449에 대한 항균활성은 첨가군에서 항균활성이 뚜렷이 나타났다(p < 0.05). 이러한 결과는 yoghurt 상등액의 낮은 pH에 의한 영향이라기 보다는 발효 중, 생성된 항균성 물질과 첨가된 감초 추출물의 영향이라고 판단된다. 발효유의 관능검사를 실시한 결과 모든 배양시간에서 첨가구들이 대조구에 비하여 낮은 점수를 받았다(p < 0.05). 이는 감초 특유의 향과 맛에 대해 익숙하지 않는 점과 과도한 산 생성으로 인해 오히려 기호도를 낮춘 것으로 사료된다. 관능검사 결과, 40% 에탄올 감초 추출물을 0.05% 함유한 yoghurt의 향, 맛, 조직감, 색택 및 전반적인 기호도가 다른 처리군에 비하여 월등히 높았다. 이에 감초 추출물을 함유한 yoghurt의 제조 시, 적절한 감초 추출물의 배합비율은 총 중량대비 0.05%인 것으로 판단된다. 식물자원에는 다양한 생리활성물질들이 서로 상호작용하면서 혼합되어 있는 형태로 다양한 질환에 대한 조합치료(combination therapy)에 이용될 수 있으며, 생체 내에서 다중 타겟에 영향을 미치므로 식물작원에 함유되어 있는 다양한 성분이 함유되어 있는 조추출물(crude extract), 용매 분획물(solvent fraction)을 의약품으로 이용하거나, 이로부터 분리된 단일성분 화합물 또는 이에 대한 유도체를 합성하여 새로운 의약품에 대한 개발이 시도되고 있다. 감초의 지표 성분이며, H. pylori에 대한 항균활성이 있는 glycyrrhizin은 비당질의 고감미에 속하는 천연 감미료로서 설탕의 약 200배의 감미를 가지고 있으며, 항산화성을 가지고 있어서 식품 성분의 산화를 방지하여 품질을 안정화시키는 작용을 하는 것으로 알려져 있기 때문에 H. pylori 억제능 이외의 부수적인 효능을 볼 수 있을 것이다. 기호성이 유지되면서 항균활성이 강화된 기능성 yoghurt의 개발을 위해서는 감초 추출물 0.2% 이상 첨가할 수 있도록 적절한 향을 조합하거나, 항균효과와 기호성을 개선할 수 있는 생약제를 이용하여 감초 추출물의 항균 활성을 강화하는 연구가 필요할 것으로 사료된다.

14-3-3 is a highly conserved, ubiquitously expressed protein family. It associates with diverse cellular proteins through its specific phosphoserine/phosphothreonine-binding activity and thus contributes to the regulation of crucial cellular processes such as metabolism, signal transduction, cell-cycle control, apoptosis, protein trafficking, transcription and stress responses. This study aims to determine changes in levels of 14-3-3 isoforms and 14-3-3 - associated proteins in Helicobacter pylori(H. pylori)-infected gastric epithelial AGS cells. AGS cells were stimulated with H. pylori(NCTC 11637) at the ratio of 300:1(bacterium:cell). Western blot analysis revealed that 14-3-3 σ was elevated at 3 hr after H. pylori treatment. Other isoforms were not significantly affected by H. pylori infection. Using immunoprecipitation to 14-3-3 σ, followed by proteomic analysis, we found that S phase kinase associated protein isoform 2 bound to 14-3-3 σ has increased. In contrast, three proteins (DEAD-box polypeptide 3, heterogeneous nuclear ribonucleoprotein H2 and WD repeat-containing protein isoform 1) bound to 14-3-3 decreased by H. pylori infection. Our results suggest that 14-3-3 may play an important regulatory role in H. pylori-induced signal transduction in gastric epithelial cells.

Lactic acid bacteria from traditional Korean foods of Tarak and Kimchi was isolated and characterized against carcinogenic Helicobacter pylori. Five Tarak and 30 Kimchi, traditional lactic acid-fermented foods, were collected from Andong area and the markets in Seoul, respectively and 15 lactic acid bacteria were isolated. Among them, two isolates were selected from high growth-inhibitory activities on H. pylori. The isolates were identified as Streptococcus thermophilus LAB kw15 from Tarak and Leuconostoc mesenteroides LAB kw5 from Kimchi by the biochemical characteristics and 16S DNA sequencing. The culture solutions of the isolates adjusted to pH 7.0 showed H. pylori inhibition. The isolates grew well and H. pylori did not grow during the co-culture with those strains. Therefore, L. mesenteroides LAB kw5 and S. thermophilus LAB kw15 might be the candidates as the functional lactic acid bacteria for improving stomach health.

Tremella fuciformis 자실체를 diethylether로 추출 후 농축하여 Muller-Hinton calf serum의 agar 배지에 100 ㎕ 씩 주입 후 4일 동안 배양된 Helicobacter pylori에 대하여 13 mm의 성장저해환을 얻었다. 이 추출물을 가지고 diethyl ether : methanol : water (1:8:1) 용매로 TLC 분리를 하여 Rf=0.95인 곳에서 12 mm, Rf=0.5에서 6 mm 그리고 Rf=0.1에서 8 mm의 저해환이 나타났으며 이것을 Waters symmetry 300 C18 5 ㎕ column으로 HPLC 분리하여 3개의 peak를 얻었 으며 이중에서 rf=0.95이 단일 peak로 나타나 순수분리 되었음을 알 수 있었다.