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Identification of Novel Psychrotolerant Bacterial Strain and Production of $\beta-Galactosidase$ (새로운 저온 내성세균의 동정과 $\beta-Galactosidase$ 생산)

  • Park, Jeong-Woon;Yoo, Jae-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.40-46
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    • 2006
  • Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.

The Ability of Cervus Elaphus Sibiricus Herbal Acupuncture to Inhibit the Generation of Inflammatory Enzymes on Collagen-induced Arthritis Mice (녹용약침(鹿茸藥鍼)이 CIA 모델 생쥐의 염증인자 생성억제에 미치는 영향)

  • Hwang, Jong-Soon;Hwang, Ji-Hye;Lee, Hyun-Jin;Lee, Dong-Gun;Kang, Min-Joo;Back, Song-Ook;Cho, Hyun-Seok;Kim, Kyung-Ho;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.1-14
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    • 2007
  • Backgrounds : Rheumatoid Arthritis(RA) is known as the chronic inflammatory diseasethat induces persistent inflammation in the joint cavity. The destruction of cartilage occurs as the result of bones destoyed by pannus, several influential cytokines induced by the synovial capsulitis, varieties of proteinases, $O_2$ radicals, and the secondary degenerative changes of articular cartilage. The type 2 collagen-induced arthritis model is used in recent experimental research on rheumatoid arthritis. Cervus elaphus sibiricus (Nockyong) has the effect of relieving pain by nourishing the muscles, joints, and bones. It is also known to be efficacious in promoting and enhancing the immune system. The objective of this study was to investigate the effect of Cervus elaphus sibiricus herbal acupuncture to inhibit the generation of proinflammatory enzyme on type 2 collagen-induced arthritis. I investigated the inhibition of mRNA transcription of MIF(macrophage migration inhibitory factor), $TNF-{\alpha}$(Tumor necrosis $factor-{\alpha}$) and MMP-9 (matrix metalloproteinase-9) of Cervus elaphus sibiricus herbal acupuncture using an in vitro test. Also investigated was the inhibition of differentiation of Th 1 cells and activation of cytokines(MIF, $TNF-{\alpha}$, IL-6, MMP-9), which are known to cause initial RA ,and are also related to the morphology of the synovial membranes of the joint capsule, by an in vivo test, using CIA(collagen induced arthritis) model mice. Materials & methods : The laboratory animals used in this experiment were 4 week-old DBA female mice, weighing approximately 20 grams, and adjusted to the laboratory environment. The experiment was divided into the normal group(NOR)-no treated group, control group(CON)-CIA induced group, and sample group(SAM)-Cervus elaphus sibiricus herbal acupuncture treated group. RA was induced in the mice via injection of $50{\mu}{\ell}$ C II mixed CFA. The Cervus elaphus sibiricus herbal acupuncture solution was applied on $GB_{35}$(陽陵泉) for 26 days from the 3rd day of RA inducement. The concentration of the solution was determined via a MTT assay. To research the effect on the expression of MIF, $TNF-{\alpha}$ and MMP-9 mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. C II induced RA knee joint's histo-chemical synovial membrane was observed using a specimen model via the Hematoxilin and Eosin dying technique. Results : The expression of mRNA of RA-related cytokines such as MIF, $TNF-{\alpha}$, and MMP-9 dosedependently decreased in the cell from the synovial membranes of the joint, which is treated with Cervus elaphus sibiricus herbal acupuncture solution. In mice treated with Cervus elaphus sibiricusherbal acupuncture, the damage of synovial membranes of the joint was lessened, and differentiation of Th 1 cells was suppressed. The activation of RA-related cytokines such as MIF was suppressed, and the generation of $TNF-{\alpha}$ and MMP-9 showed a statistically significant decreas. Conclusions : It is speculated that Cervus elaphus sibiricus herbal acupuncture has the therapeutic effect of palliating the damage of the tissue impaired by RA by inhibition of the initial RA progression and by regulating excessive differentiation of Th 1 cell as it suppresses the generation of RA-related cytokines during the highest stage of RA by acting on pro-inflammatory enzymes.

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Association of Cytotoxic T-lymphocyte Antigen-4 Polymorphisms with Malignant Bone Tumors Risk: A Meta-analysis

  • Zhang, Chao;Hou, Wei-Hua;Ding, Xuan-Xi;Wang, Xiong;Zhao, Hui;Han, Xing-Wen;Wang, Wen-Ji
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3785-3791
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    • 2016
  • Background: Previous studies have assessed the association between the Cytotoxic T-lymphocyte Antigen-4(CTLA-4) polymorphism with the risk of malignant bone tumor, but the conclusions were inconsistent. We aimed to clarify association of cytotoxic T-lymphocyte antigen-4 polymorphisms with malignant bone tumors risk by performing a meta-analysis. Materials and Methods: The databases including PubMed, EMBase databases and the Cochrane Library were searched to identify the eligible studies prior to January 30 2016. Odds ratio (OR) with 95% confidence interval (95%CI) were used to estimate the strengths of the association between the CTLA-4 polymorphism and the malignant bone tumor risks. The meta-analysis was performed by STATA 12.0. Results: Four individual studies with a total of 1003 cases with malignant bone tumor and 1162 controls were included in our meta-analysis. The results of meta-analysis on those data demonstrated that CTLA-4 +49G>A polymorphism was associated with the risk of Ewing's sarcoma and osteosarcoma strongly (A vs. G: OR=1.36, 95%CI:1.20-1.54, p=0.000; AA+AG vs. GG: OR=1.35, 95%CI:1.14-1.61, p=0.001; AA vs. GG: OR=2.24, 95%CI:1.67-2.99, p=0.000; AA vs. AG+GG: OR=2.00, 95%CI:1.53-2.62, p=0.000), but CTLA-4 -318C/T polymorphism was not associated with the risk of malignant bone tumor (C vs. T: OR=0.76, 95%CI:0.76-1.08, p= 0.262; CC+CT vs. TT: OR=0.70, 95%CI:0.41-1.20, p= 0.198; CC vs. TT: OR=0.69, 95%CI:0.40-1.19, p= 0.183; CC vs. CT+TT: OR=0.92, 95%CI:0.75-1.13, p= 0.419). Subgroup analysis showed that there are significantly positive correlations between CTLA-4 +49G>A polymorphism and increased risks of malignant bone tumors in large size of sample (A vs. G: OR=1.347, 95%CI: 1.172,1.548, p=0.000; AA vs. GG: OR=2.228, 95%CI: 1.608,3.085, p=0.000), Ewing's Sarcoma or Osteosarcoma (A vs. G: OR=1.361, 95%CI: 1.201,1.540, p=0.000; AA vs. GG: OR=2.236, 95%CI: 1.674,2.986, p=0.000), and PCR-RFLP or Sequencing(A vs. G: OR=1.361, 95%CI: 1.201,1.540, p=0.000; AA vs. GG: OR=2.236, 95%CI: 1.674,2.986, p=0.000), but CTLA-4 -318C/T polymorphism was not associated with the risk of malignant bone tumors in diagnosis, genotype method, and sample size (all p>0.05). Conclusions: CTLA-4 +49A/G variant was associated with an increased risk of developing the malignant bone tumors, such as Ewing's sarcoma and osteosarcoma. However, it failed to show the association between CTLA-4 -318C/T polymorphism and the risk of malignant bone tumors. Future large-scale studies remain to be done to confirm our conclusions.

Association between Antipsychotic-Induced Restless Legs Syndrome and Glutathione S-Transferase Gst-M1, Gst-T1 and Gst-P1 Gene Polymorphisms (Glutathione S-Transferase (GST) 유전자 다형성과 항정신병약물로 유발된 하지불안증후군의 연관 연구)

  • Kang, Seung-Gul;Park, Young-Min;Kim, Leen;Lee, Heon-Jeong
    • Sleep Medicine and Psychophysiology
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    • v.22 no.1
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    • pp.25-29
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    • 2015
  • Objectives: The pathophysiology of restless legs syndrome (RLS) has not been fully elucidated. Oxidative stress might play a role in the development of RLS and other antipsychotic-induced side effects such as tardive dyskinesia. In the present study, we investigated whether the glutathione S-transferase (GST) gene polymorphisms are associated with antipsychotic-induced RLS in schizophrenia. Methods: We assessed antipsychotic-induced RLS symptoms in 190 Korean schizophrenic patients using the diagnostic criteria of the International Restless Legs Syndrome Study Group. The GST-M1, GST-T1 and GST-P1 loci were analyzed using PCR-based methods. Results: We divided the subjects into 2 groups: those with RLS symptoms (n = 96) and those without RLS symptoms (n = 94). There were no significant differences in the distributions of the GST-M1 genotypes (${\chi}^2=3.56$, p = 0.059), GST-T1 (${\chi}^2=0.51$, p = 0.476) and GST-P1 (${\chi}^2=0.57$, p = 0.821) between the 2 groups. Comparison of the RLS score among genotypes of the GST-M1 (t = -1.54, p = 0.125), GST-T1 (t = -0.02, p = 0.985) and GST-P1 (F = 0.58, p = 0.560) revealed no significant difference. Conclusion: These data suggest that GST gene polymorphisms do not confer increased susceptibility to RLS symptoms in schizophrenic patients. Future studies are necessary to evaluate the possible influences of other candidate genes involved in the reactive oxygen species system.

Study on the Clonality of Endothelial Cell Proliferation in Plexiform Lesions in Patients with Pulmonary Hypertension Associated with CREST Syndrome (CREST 증후군에 동반된 폐고혈압 환자에서 총상병변내 내피세포 증식의 클론성에 관한 연구)

  • Lee, Sang-Do;Jeon, Yong-Gam;Lee, Ji-Hyun;Shim, Tae-Sun;Lim, Chae-Man;Koh, Yun-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Tuder, Rubin M.
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.2
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    • pp.150-160
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    • 1999
  • Background: The CREST syndrome is an indolent form of progressive systemic sclerosis. Although its clinical progress is indolent, pulmonary hypertension(PH) associated with CREST syndrome have grave prognosis with over 40 percent mortality rate at 2 year follow-up. But the pathogenesis of pulmonary hypertension in this disease is not known, and classified as either primary or secondary PH. Clonality of endothelial cell proliferation in plexiform lesion is a molecular marker which allows distinction between primary and secondary PH. We performed this study to know whether the PH associated with CREST syndrome is a variant of primary PH or is a secondary PH. Methods: We assessed the X-chromosome inactivation based on the methylation pattern of the human androgen-receptor gene by PCR(HUMARA). Endothelial cells in plexiform lesions from female patients(n=3) with PH associated with CREST syndrome were microdissected from paraffin blocks. Vascular smooth muscle cells and lung parenchyma were also microdissected for clonality studies. Results: The proliferating endothelial cells in 14 plexiform lesions were all polyclonal. Similarly proliferated smooth muscle cells from 5 vessels with medial hypertrophy were also polyclonal. Conclusion: These results suggest that the pulmonary hypertension associated with CREST syndrome has different pathogenesis from primary PH and to be classified as secondary PH.

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Loss of Heterozygosity on the Long Arm of Chromosome 21 in Non-Small Cell Lung Cancer (비소세포폐암에서 21q 이형체 소실)

  • Chai, Po-Hee;Bae, Nack-Cheon;Lee, Eung-Bae;Park, Jae-Yong;Kang, Kyung-Hee;Kim, Kyung-Rok;Bae, Moon-Seob;Cha, Seung-Ik;Chae, Sang-Chul;Kim, Chang-Ho;Jung, Tae-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.6
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    • pp.668-675
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    • 2001
  • Background : Non-smalll lung cancer(NSCLC) develops as a result of the accumulation of multiple genetic abnormalities. Loss of heterozygosity(LOH) is one of the most frequent genetic alterations that is found in NSCLC, and the chromosomal regions that display a high rate of LOH are thought to harbor tumor suppressor genes(TSGs). This study was done to determine the frequency of LOH in 21q with the aim of identifying potential TSG loci. Method : Thirty-nine surgically resected NSCLCs were analysed. Patients peripheral lymphocytes were used as the source of the normal DNA. Five microsatellite Inarkers of 21q were used to study LOH : 21q21.1(D21S1432, and D21S1994); 21q21.2-21.3(D21S1442) ; 21q22.1(21S1445) ; and 21q22.2-22.3(D21S266). The fractional allelic loss(FAL) in a tumor was calculated as the ratio of the number of markers showing LOH to the number of informative markers. Result : LOH for at least one locus was detected in 21 of 39 tumors(53.8%). Among the 21 tumors with LOH, 5(21.8%) showed LOH at almost all informative loci. Although statistically not significant, LOH was found more frequently in squamous cell carcinomas(15 of 23, 65.2%) than in adenocarcinomas(6 of 16, 37.5%). In the squamous cell carcinomas the frequency of LOH was higher in stage II-III (80.0%) than in stage I (53.8%). The FAL value in squamous cell carcinomas($0.431{\pm}0.375$) was significantly higher than that found in adenocarcinomas($0.l92{\pm}0.276$). Conclusion : These results suggest that LOH on 21q may be involved in the development of NSCLC, and that TSG(s) that contribute to the pathogenesis of NSCLC may exist on 21q.

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Clinical Significance of Low-colony Count Scotochromogen Nontuberculous Mycobacteria (균집락수가 적은 암색소성 비결핵항산균 배양의 임상적 의미)

  • Lee, Jung Yeon;Kim, Mi-Na;Chung, Hee-Jung;Jun, Kyung Ran;Choi, Hee Jin;Lee, Hyeyoung;Joung, Eun Young;Oh, Yeon-Mok;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Shim, Tae Sun
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.1
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    • pp.39-46
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    • 2005
  • Background : Even though it has been suggested that low-colony, scotochromogen nontuberculous mycobacteria (NTM) are usually contaminants and not true pathogens, evidence for this hypothesis has not been provided. This study investigated the colony characteristics, organism identification, and clinical significance of low-colony scotochromogen. Methods : The laboratory cultured 6,898 respiratory clinical specimens for an examination of mycobacteria over a three-month period. A low-colony count was arbitrarily defined as ${\leq}20$ colonies. This study analyzed the recovery rate of the mycobacteria, the number of colonies and their gross characteristics, and their clinical significance. PCR-restriction fragment length polymorphism analysis was carried out to identify the NTM species. NTM pulmonary disease was defined according to the American Thoracic Society. Results : A total of 6,898 respiratory specimens for mycobacterium were cultured. Of these, 263 (3.8%) grew NTM, and 382 (5.5%) grew M. tuberculosis. Of the 263 cultured NTM specimens, 124 (47.1%) were scotochromogens. The smear-positive rate was significantly lower in these scotochromogens (4.8%) than in the non-scotochromogens (23.7%) (p<0.05). The most common isolates were M. gordonae (83/102, 81.4%) in the scotochromogens, and MAC (52/121, 43.0%) in the non-scotochromogens. Even though three out of 113 patients with a low-colony scotochromogen has been diagnosed with NTM pulmonary disease, the isolated scotochromogen was not considered to be the cause of the NTM disease but was just a contaminant. Conclusion : In this study, the most common isolate of a low-colony count scotochromogen was M. gordonae, which appeared to be contaminants and not true pathogens. Greater efforts in the quality control of a mycobacterium laboratory are needed in cases where there is a high recovery rate of low-colony count scotochromogen.

STIMULATION OF OSTEOBLASTIC PHENOTYPES BY STRONTIUM IN PERIOSTEAL-DERIVED CELLS (골막기원세포에서 strontium에 의한 조골세포 표현형의 활성)

  • Kim, Shin-Won;Kim, Uk-Kyu;Park, Bong-Wook;Hah, Young-Sool;Cho, Hee-Young;Kim, Jung-Hwan;Kim, Deok-Ryong;Kim, Jong-Ryoul;Joo, Hyun-Ho;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.3
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    • pp.199-206
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    • 2010
  • This study investigated the effects of strontium on osteoblastic phenotypes of cultured human periostealderived cells. Periosteal tissues were harvested from mandible during surgical extraction of lower impacted third molar. Periosteal-derived cells were introduced into cell culture. After passage 3, the periostealderived cells were further cultured for 28 days in an osteogenic induction DMEM medium supplemented with fetal bovine serum, ascorbic acid 2-phosphate, dexamethasone and at a density of $3{\times}10^4$ cells/well in a 6-well plate. In this culture medium, strontium at different concentrations (1, 5, 10, and 100 ${\mu}g$/mL) was added. The medium was changed every 3 days during the incubation period. We examined the cellular proliferation, histochemical detection and biochemical measurements of alkaline phosphatase (ALP), the RT-PCR analysis for ALP and osteocalcin, and von Kossa staining and calcium contents in the periostealderived cells. Cell proliferation was not associated with the addition of strontium in periosteal-derived cells. The ALP activity in the periosteal-derived cells was higher in 5, 10, and 100 ${\mu}g$/ml strontium-treated cells than in untreated cells at day 14 of culture. Among the strontium-treated cells, the ALP activity was appreciably higher in 100 ${\mu}g$/ml strontium-treated cells than in 5 and 10 ${\mu}g$/ml strontium-treated cells. The levels of ALP and osteocalcin mRNA in the periosteal-derived cells was also higher in strontium-treated cells than in untreated cells at day 14 of culture. Their levels were increased in a dose-dependent manner. Von Kossa-positive mineralization nodules were strongly observed in the 1 ${\mu}g$/ml strontium-treated cells at day 21 and 28 of culture. The calcium content in the periosteal-derived cells was also higher in 1 ${\mu}g$/ml strontium-treated cells at day 28 of culture. These results suggest that low concentration of strontium stimulates the osteoblastic phenotypes of more differentiated periosteal-derived cells, whereas high concentration of strontium stimulates the osteoblastic phenotypes of less differentiated periosteal-derived cells. The effects of strontium on osteoblastic phenotypes of periosteal-derived cells appear to be associated with differentiation-extent.

Genetic Variation of some Patches of Eleutherococcus senticosus (Rupr. & Maxim) Maxim. in Korea (국내(國內) 가시오갈피 군락(群落)의 유전변이(遺傳變異) 분석(分析))

  • Hong, Kyung-Nak;Cho, Kyung-Jin;Park, Yew-Heon;Hur, Sung-Du;Hong, Yong-Pyo;Kang, Bum-Yong
    • Journal of Korean Society of Forest Science
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    • v.89 no.5
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    • pp.645-654
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    • 2000
  • The aim of this study was to described the genetic structure of Eleutherococcus senticosus in Korea. We investigated 10 patches, which are eight Korean patches and two foreign patches come from Russia and China growing at Korean habitat, using ISSR(inter-simple sequence repeats) markers. In ISSR PCR, the overall percentage of polymorphic ISSR amplicons was 76% and the mean number of amplicons per ISSR primer was 11.5, which were higher than the RAPD results for the some cultivars collected in Korea(Kim et al., 1998) ; 57% and 5.7, respectively. So ISSR markers provide more powerful tool than RAPD markers for the investigation of genetic variation in E. senticosus. There are relatively high genetic variation among patches as 62.8%, but low variation within eight Korean patches. Such pattern of genetic variation, which is not ordinary in other tree species, may be result from the narrow and limited habitats and the asexual reproduction of this species at the natural stands in Korea. Although the small sample size in this study seemed to be resulted in the high genetic variation among patches, the overall genetic interpretation of this study might not be much affected on the basis of the characteristics of the distribution and the reproduction system of E. senticosus. Analysis of genetic distance between all pairs of the patches did not reveal any trends with regard to geographic distance, which was confirmed by the results obtained from AMOVA(analysis of molecular variance) and PCA(principal component analysis). These results suggest that, in addition to the preservation of the natural stands, the conservation of larger number of patches with small number of individuals per patch is more effective for the ex situ conservation and for maintaining the genetic diversity of E. senticosus in Korea than smaller patches with large number of individuals.

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Inhibitory Effects of Chios Mastic Gum on Gastric Acid Secretion by Histamine-Related Pathway in a Rat Model and Primary Parietal Cells (위염 동물모델과 위 벽세포에서 히스타민 경로를 통한 매스틱검(Chios Mastic Gum)의 위산 분비 억제효과 및 기전 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Shim, Tae Jin;Lee, Jum Kyun;Hwang, Kwon-Tack
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1500-1509
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    • 2014
  • The object of this study was to investigate the inhibitory effects of chios mastic gum (MG) on gastric acid secretion in an ethanol-induced SD rat model and primary parietal cells. Rats were randomly divided into four groups: Vehicle (normal group), Control (treated with ethanol), MG50 (treated with ethanol and mastic gum at 50 mg/kg b.w), MG100 (treated with ethanol and mastic gum at 100 mg/kg b.w). Groups treated with both MG50 and MG100 showed attenuation of gastric mucosal injury, sub-epithelial loss, hemorrhaging, and gastric juice secretion. We also examined the acidity of gastric juice during gastric injury. Oral administration of both MG50 and MG100 significantly decreased acidity of gastric juice by % and %, respectively. To examine the stimulatory factors related to gastric acid secretion, mRNA expression levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured by real-time PCR. Compared with a vehicle group, mRNA expression levels of H2r, CCK2r, and $H^+/K^+$ ATPase clearly increased in the control group. However, levels of H2r, CCK2r, and $H^+/K^+$ ATPase slightly but significantly decreased in MG-treated groups compared with control. Blood level of histamine significantly decreased in MG-treated groups, which indicates the involvement of MG on in histamine-related acid secretion. To identify the mode of action of MG in regulating histamine-related pathways, intracellular level of cAMP and mRNA levels of H2r, M3r, CCK2r, and $H^+/K^+$ ATPase were measured in primary parietal cells. While mRNA levels of M3r and CCK2r remained unchanged, levels of H2r and $H^+/K^+$ ATPase significantly decreased upon MG treatment. Subsequently, intracellular levels of cAMP decreased. These results suggest that mastic gum has the ability to inhibit gastric acid secretion by regulating a histamine-related pathway.