• Title/Summary/Keyword: P19 cells

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The Effect of Inchinohryungsan Pharmacopuncture on Hepatocellular Carcinoma Induced by Diethylnitrosamine(DEN) in Rats (인진오령산(茵陳五苓散)약침이 DEN으로 유발된 흰쥐의 간암에 미치는 영향)

  • Ryu, Mi-Seon;Kim, Sung-Phil;Kim, Jae-Hong;Ryu, Hye-Seon;Yoon, Yeo-Choong;Shin, Jeong-Cheol
    • Journal of Acupuncture Research
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    • v.28 no.5
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    • pp.39-55
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    • 2011
  • Objectives : Inchinohryungsan has been used for treatment of hepatobiliary diseases. This study was designed to investigate the effect of Inchinohryungsan pharmacopuncture on hepatocellular carcinoma in rats. Sprague Dawley(SD) rats of the control and experimental groups received intraperitoneal injection of 50 mg/kg DEN, weekly for 12 weeks. Methods : Rats were divided into 5 groups. Normal group was not induced hepatocellular carcinoma and not treated. Control group was induced hepatocellular carcinoma and injected with Inchinohryungsan pharmacopuncture into the root of tail. Experimental groups were induced hepatocellular carcinoma. BL group was injected with Inchinohryungsan pharmacopuncture into the $BL_{18}$ and $LR_{14}$, BG group was injected into the $BL_{19}$ and $GB_{24}$ and CSC group was injected into the $CV_{12}$, $ST_{25}$ and $CV_4$. Thereafter, the changes of the body weight, the liver weight and the weight of liver/100g body weight, WBC, neutrophil, lymphocyte and the activities of AST, ALT, ALP, LDH, AFP and SOD were measured. And gross anatomy, light and electron microscopy were performed. Results : The significant results were as follows, 1. The activities of LDH were significantly decreased in CSC group compared with control group. 2. The activities of AFP were significantly decreased in the BL, BG, CSC groups compared with control group. 3. The activities of SOD were increased in the BL, BG, CSC groups compared with control group and CSC group was significantly increased than normal group. 4. According to the gross anatomical observation, the control and BL, BG, CSC groups showed multi-nodular hepatocellular carcinoma. But the size and numbers of the hepatocellular carcinoma in experimental groups were smaller than control group. 5. The numbers of hepatic p53 positive cells were decreased in the BL, BG groups compared with control group. 6. According to the light and electron microscopical observation, the BL, BG and CSC groups were mildly improved than control group in morphological and histopathological changes. Conclusions : These results suggested that Inchinohryungsan pharmacopuncture may have some effects on hepatocellular carcinoma induced by DEN in rats.

Evaluation of In-vitro Anticoagulation Activity of 33 Different Medicinal Herbs (33종 생약재의 in-vitro 항혈전 활성 평가)

  • Ryu, Hee-Young;Ahn, Seon-Mi;Kim, Jong-Sik;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.20 no.6
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    • pp.922-928
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    • 2010
  • This study was carried out to develop safe and novel anticoagulation agents from oriental medicinal herbs. From 33 medicinal herbs, 40 different ethanol extracts were prepared according to place of origin or extraction parts, and anticoagulation activities were evaluated by determination of thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (aPTT). The average water content and average extraction ratio for the medicinal herbs were $6.85{\pm}2.26%$ and $5.27{\pm}4.25%$, respectively. Evaluation of TT at various concentrations of the extract led to the selection of Mucuna birdwoodiana, Prunus armeniaca, Cacalia ainsliaeiflora, Cinnamonum aromaticum, and Rhus javanica Linneas potent antithrombosis medicinal herbs. Evaluation of PT and aPTT showed that the extracts of R.javanica Linne, M. birdwoodiana, and P. armeniaca have strong anticoagulation activities. Determination of hemolytic activities of 40 different ethanol extracts against human red blood cells, however, showed that only M. birdwoodiana, C. ainsliaeiflora, C. aromaticum, and R. javanica Linnehas strong anticoagulation activity without hemolytic activity at a concentration of 500 mg/ml. Our results suggest that oriental medicinal herbs, which are under a mass-production system, have potentialas a safe and novel source of anticoagulants, as well being a thrombin-specific and coagulation factor-specific inhibitor.

The Regenerative effects of Platelet-Rich Plasma and Enamel Matrix Protein on Grade III Furcation defects in beagle dogs (혈소판 농축혈장과 법랑기질 단백질이 성견 3급 이개부 병소의 재생에 미치는 영향)

  • Kim, Young-Jun;Lim, Sung-Bin;Chung, Chin-Hyung;Hong, Ki-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.823-837
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    • 2005
  • The purpose of this study was to study the histopathological correlation between the use of platelet-rich plasma and enamel matrix protein used in conjunction with xenograft. compared to a control group with regards to bone regeneration at the grade III furcation area in beagle dogs. Control group was treated with bovine derived bone $powder(Biocera^{(R)})$, and experimental I group was treated with bovine derived bone powder and Platelet-rich plasma and experimental II group was treated with bovine derived bone powder and Enamel matrix $protein(Emdogain^{(R)})$. The regeneration rate of bone formation was observed and compared histopathologically at 2. 4, and 8 weeks after surgery. The results were as follows: 1. In control group and both experimental groups. inflammatory cells were observed but, new bone formation wasn't. 2. In control group, new cementum on the notch was found in 4 weeks, less mature periodontal ligament when compared to that of experimental group was found and cementum formation was great but, regeneration couldn't be seen in 8 weeks. 3. Experimental I group. new bone formation in the area adjacent to alveolar bone and graft material surrounded by more dense connective tissue were found in 4 weeks. New bone formation up to crown portion was found and periodontal ligament was aligned functionally and cementum more mature. 4. Experimental II group, new bone formation was found under the defect area in 4 weeks and new bone formation around graft material in 8 weeks, too, and there were a number of fibroblasts, blood vessels, acellular cementum, which was less mature when compared to that of experimental I group, and dense collagen fiber like which normal periodontal ligament has in periodontal ligament of experimental II group in 8 weeks. 5. As a result of histologic finding, bone formation rate were 18.0${\pm}$7.87%(control group), 34. 05${pm}$7.25%(experimental I group), 19.33 ${pm}$5.15%(experimental II group) in 4 weeks and 21.89${pm}$1.58%(control group), 38.82${pm}$3.2(experimental I group), 37.65${pm}$9.22%(experimental II group) in 8 weeks. 6. Statistically significant ratio of bone formation was observed in experimental I group in 4 weeks and in experimental II group in 8 weeks. When experimental I group was compared to experimental II group, the ratio of bone formation in experimental I group was higher than that in experimental II group in 4 weeks(p<0.05). This results suggest that platelet-rich plasma showed more new bone formation than enamel matrix protein within 4 weeks. And use of enamel matrix protein in the treatment of periodontal bone defects starts to enhance regeneration after 8 weeks in beagle dogs.

Effects of Red Ginseng Total Saponin on the Polycystic Ovaries in Rats (홍삼 사포닌이 랫드의 다낭성난소에 미치는 영향)

  • Lee Jiae;Lim Sung-Chul;Lyu Ae-Ran;Bae Jin-Gyu;Kang Seong-Soo;Kim Jong-Choon;Kim Sung-Ho;Kim Jeong-Wook;Choi Bum-Chae;Bae Chun-Sik
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.433-436
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    • 2004
  • Polycystic ovarian syndrome (PCOS) is a very common endocrine disorder in women of reproductive age. There are some evidences that nerve growth factor (NGF) is involved in the pathogenesis of PCOS. In this study, we investigated the effect of Korean red ginseng total saponin (GTS) on the ovarian morphology and NGF expressions in the ovaries, pituitary and hippocampus. The oil control animals were injected with 0.2 ml oil/rat. Animals in estradiol valerate (EV) control group were injected i.m. with 4 mg EV in 0.2 ml oil/rat. The GTS was administered (100 mg/kg) i.p. every other day for 60 days, beginning 1 day after the EV injection. PCO was induced by a single injection of EV (4 mg, i.m.). At day 60, the expressions of NGF were examined by immunohistochemistry. The main findings were as follows; PCO was fully developed with a single i.m. injection of EV, and PCO showed the increased expression of NGF, and GTS administration decreased NGF expressions in the ovaries without affecting pituitary and hippocampus significantly. The present results demonstrate that GTS attenuates PCOS by the stimulation of NGF expression.

Drying seaweeds using hybrid hot water Goodle dryer (HHGD): comparison with freeze-dryer in chemical composition and antioxidant activity

  • Nagahawatta, D.P.;Asanka Sanjeewa, K.K.;Jayawardena, Thilina U.;Kim, Hyun-Soo;Yang, Hye-Won;Jiang, Yunfei;Je, Jun-Geon;Lee, Tae-Ki;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.24 no.1
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    • pp.19-31
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    • 2021
  • Seaweeds are a potential source of minerals, essential amino acids, fatty acids, proteins, and various bioactive compounds such as antioxidants. The higher water content of seaweeds reduces the shelf life and this requires the appropriate drying method. The drying conditions play a major role in the conservation of nutrient composition in dried seaweeds. In recent years, the seaweed industry has used many different drying methods with advantages and limitations. Hybrid hot-water Goodle dryer (HHGD) which is a special dryer mixed with hot-water and a Korean traditional heating system (Goodlejang) might be a solution to avoid these limitations. The present study evaluated the effect of drying conditions in HHGD on nutrient composition and bioactivities of brown seaweeds. Moreover, freeze-dryer (FD) and HHGD were employed in this study to compare the dried outputs obtained from four brown seaweed species. The present study aims to evaluate the effect of the hybrid hot-water Goodle drying method (HHGDM) on the nutritional composition and antioxidant activity of dried seaweeds. AOAC standard methods were used to analyze the proximate composition of dried samples and their 70% ethanol extract. The intracellular and extracellular antioxidant activities were evaluated using Vero cells and electron spin resonance (ESR) spectrometer respectively. High performance liquid chromatography, apoptotic body formation, and in-vivo experiments were used for further confirmation of the quality of dried output. The proximate composition results obtained from drying in HHGD and FD did not exhibit any significant difference. Moreover, the seaweed extracts from the dried seaweeds by HHGD and FD dryings were also not different and both significantly down-regulated in-vivo and in-vitro oxidative stress. Furthermore, the high performance liquid chromatography results revealed that the two dryers did not make the major peaks different in the chromatograms. Freeze-drying method (FDM) provides elevated quality for dried output, but there are limitations such as high cost and low capacity. The results from a novel HHGD did not provide any significant difference with the results in FD and expressed a potential to avoid the limitations in FD. Overall, these findings solidified the applicability of HHGD over FD.

Expression of TIMP1, TIMP2 Genes by Ionizing Radiation (이온화 방사선에 의한 TIMP1, TIMP2 유전자 발현 측정)

  • Park Kun-Koo;Jin Jung Sun;Park Ki Yong;Lee Yun Hee;Kim Sang Yoon;Noh Young Ju;Ahn Seung Do;Kim Jong Hoon;Choi Eun Kyung;Chang Hyesook
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.171-180
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    • 2001
  • Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

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Correlation of Proliferating Cell Nuclear Antigen (PCNA) Expression and S-phase Fraction, Survival Rate in Primary Non-Small Cell Lung Cancer (원발성 비소세포 폐암에서 PCNA의 발현정도와 암세포의 분열능 및 생존률과의 관계)

  • Yang, Sei-Hoon;Kim, Hak-Ryul;Gu, Ki-Seon;Jung, Byung-Hak;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.756-765
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    • 1997
  • Background : To study the prognosis of patients with lung cancer, many investigators have reported the methods to detect cell proliferation in tissues including PCNA, thymidine autoradiography, flow cytometry and Ki-67. PCNA, also known as cyclin, is a cell related nuclear protein with 36KD intranuclear polypeptide that is maximally elevated in S phase of proliferating cells. In this study, PCNA was identified by paraffin-embedding tissue using immunohistochemistry which has an advantage of simplicity and maintenance of tissue architecture. The variation of PCNA expression is known to be related with proliferating fraction, histologic type, anatomic(TNM) stage, degree of cell differentiation, S-phase fraction and survival rate. We analyzed the correlation between PCNA expression and S-phase fraction, survival. Method : To investigate expression of PCNA in primary lung cancer, we used immunohistochemical stain to paraffin-embedded sections of 57 resected primary non-small cell lung cancer specimen and the results were analyzed according to the cell type, cell differentiation, TNM stage, S-phase fraction and survival. Results : PCNA expression was divided into five group according to degree of staging(-, +, ++, +++, ++++). Squamous cell type showed high positivity than in adenocarcinoma. Nonsignificant difference related to TNM stage was noticed. Nonsignificant difference related to degree of cell differentiation was noticed. S-phase fraction was increased with advance of PCNA positivity, but it could not reach the statistic significance. The 2 year survival rate and median survival time were -50% 13 months, +75% 41.3 months, ++73% 33.6 months, +++67% 29.0 months, ++++25% 9 months with statistic significance (P<0.05, Kaplan-Meier, generalized Wilcox). Conclusion : From this study, PCNA expression was high positive in squamous cell cancer. And, there was no relationship between PCNA positivity and TNM stage, cellular differentiation or S-phase fraction. But, the patients with high positive PCNA staining showed poor survival rate than the patients with lower positive PCNA staining (p<0.05). It was concluded that PCNA immunostaining is a simple and useful method for survival prediction in paraffin embedded tissue of non-small cell lung cancer.

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Association between polymorphisms in Interleukin-17 receptor A gene and childhood IgA nephropathy (IgA 신병증 환자에서 Interleukin-17 수용체 A 유전자의 단일염기다형성 연관성 연구)

  • Baek, Seung-Ah;Han, Won-Ho;Cho, Byoung-Soo;Kim, Sung-Do
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.215-221
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    • 2010
  • Purpose : Interleukin-17 (IL-17) is produced by activated CD4+T cells and exhibits pleiotropic biological activity on various cell types. IL-17 was reported to be involved in the immunoregulatory response in IgA nephropathy (IgAN). Our aim was to investigate the association between single-nucleotide polymorphisms (SNPs) in IL-17 receptor A (IL-17RA) gene and childhood IgAN. Methods : We analyzed the SNPs in the IL-17RA in 156 children with biopsy-proven IgAN and 245 healthy controls. We divided the IgAN patients into 2 groups and compared them with respect to proteinuria (${\leq}4$ and >$4mg/m^2/h$, ${\leq}40$ and >$40mg/m^2/h$, respectively) and the presence of pathological levels of biomarkers of diseases such as interstitial fibrosis, tubular atrophy, or global sclerosis. Results : No difference was observed between the SNP genotypes rs2895332, rs1468488, and rs4819553 between IgAN patients and control subjects. In addition, no significant difference was observed between allele frequency of SNPs rs2895 332, rs1468488, and rs4819553 between patients in the early and advanced stage of the disease. However, significant difference was observed between the genotype of SNP rs2895332 between patients with proteinuria (>$4mg/m^2/h$) and those without proteinuria (codominant model OR 0.36, 95% CI 0.19-.66, P <0.001; dominant model OR 0.35, 95% CI 0.17-.69 P =0.002; recessive model OR 0.12, 95% CI 0.01-.06 P =0.025). Conclusion : Our results indicate that the SNP in IL-17RA (rs2895332) may be related to the development of proteinuria in IgAN patients.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Characteristics and anti-obesity effect of fermented products of coffee wine (커피발효물의 발효특성 및 항비만 효과)

  • So Hyun Park;Hyeon Hwa Oh;Do Youn Jeong;Young-Soo Kim
    • Food Science and Preservation
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    • v.30 no.4
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    • pp.703-715
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    • 2023
  • This study was conducted to investigate the fermentation characteristics and anti-obesity effects of acetic acid fermentation products of coffee wine. The live cell counts, soluble solids, pH and total acidity of the acetic acid unfermented coffee wine (AUFCW; day 0, before fermentation) were 6.35 log CFU/mL, 8.10 °Brix, 3.88, and 1.29%, respectively, while the acetic acid fermented coffee wine (AFCW; day 15, after fermentation) were 4.40 log CFU/mL, 8.57 °Brix, 3.07, and 7.45%, respectively. Pancreatic lipase inhibitory activity tended to increase as the acetic acid fermentation period increased. The anti-obesity effects of AFCW on 3T3-L1 cells, which was induced by MDI, were evaluated based on the lipid accumulation rate, leptin expression, and fat production-related gene expression (PPAR-γ and SREBP-1c) at the mRNA level. In the case of AFCW, the lipid accumulation rate and leptin expression were decreased to 69.37% and 50.20% at a concentration of 200 ㎍/mL, respectively, and the expression levels of PPAR-γ and SREBP-1c at the mRNA level were decreased to 79.89% and 48.81%, respectively. These results indicate that anti-obesity effect of acetic acid fermentation products could be increased by acetic acid fermentation of coffee wine.