• 한국미생물·생명공학회지
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• 제39권3호
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• pp.266-273
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• 2011

인도산 울금 분말을 bifidobacteria와 lactobacilli를 포함한 프로바이오틱 유산균으로 발효한 시료들의 소염활성 정도를 세포 내 염증성 인자들의 발현양을 관찰함으로써 평가하였다. 그 중 L. johnsonii IDCC 9203으로 발효한 시료의 소염 활성이 가장 뛰어났다. 이를 바탕으로 울금 분말을 포함한 본배양 배지에서 L. johnsonii IDCC 9203으로 21시간 동안 배양 후 얻은 상등액으로 발효 울금 원료를 제조하였고, 제조된 발효 울금 원료에 대한 소염활성 효능을 테스트하기 위해 LPS로 활성화된 raw 264.7 세포주에 처리하고 COX-2와 iNOS의 발현양을 확인하였다. 그 결과 발효 울금 원료 250 ${\mu}g$/mL까지 농도 의존적으로 COX-2와 iNOS의 발현을 감소시켰으며, 그 저해 활성은 동일 농도의 비발효 울금 원료보다 강하였다. NC/Nga 아토피 피부염 동물모델과 PCA 동물모델에서 발효 울금 원료의 효능 확인 결과 대조군에 비해 아토피 피부염의 초기 증상 개선효과와 급작형 과민반응에 대한 예방효과가 뛰어남을 확인하였다. 발효 울금 원료의 유효성분 함량을 분석했을 때 커큐민의 함량은 비발효 울금 원료에 비해 2.5배 증가했으며, 수용성 커큐민의 함량 역시 증가하였다. 또한 비스디메톡시커큐민이나 디메톡시커큐민의 함량도 증가되었을 뿐 아니라 전체 커큐미노이드 중에서 이들 유도체의 비중이 높아짐을 확인하였다. 모든 결과들을 종합하면, 울금 분말을 L. johnsonii IDCC 9203을 이용하여 발효함으로써 유효성분인 커큐미노이드들의 성분비가 변화하고 수용성 커큐미노이드의 증가에 의한 생체 이용율 증가로 울금의 소염 및 항알레르기 활성이 증가된다. 본 연구를 통해 L. johnsonii IDCC 9203으로 발효한 울금 원료는 급성기 피부염에 대한 예방 및 치료 목적으로 사용 가능할 것으로 예상된다.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.1022-1029
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• 2018

Tuberculosis (TB) remains a serious health issue around the word. Adenovirus (Ad)-based vaccine and modified vaccinia virus Ankara (MVA)-based vaccine have emerged as two of the most promising immunization candidates over the past few years. However, the performance of the homologous and heterologous prime-boost immunization regimens of these two viral vector-based vaccines remains unclear. In the present study, we constructed recombinant Ad and MVA expressing an Ag85B-TB10.4 fusion protein (AdH4 and MVAH4) and evaluated the impact of their different immunization regimens on the humoral and cellular immune responses. We found that the viral vector-based vaccines could generate significantly higher levels of antigen-specific antibodies, $IFN-{\gamma}$-producing splenocytes, $CD69^+CD8^+$ T cells, and $IFN-{\gamma}$ secretion when compared with bacillus Calmette-$Gu{\acute{e}}rin$ (BCG) in a mouse model. AdH4-containing immunization regimens (AdH4-AdH4, AdH4-MVAH4, and MVAH4-AdH4) induced significantly stronger antibody responses, much more $IFN-{\gamma}$-producing splenocytes and $CD69^+CD8^+$ T cells, and higher levels of $IFN-{\gamma}$ secretion when compared with the MVAH4-MVAH4 immunization regimen. The number of $IFN-{\gamma}$-producing splenocytes sensitive to $CD8^+$ T-cell restricted peptides of Ag85B (9-1p and 9-2p) and Th1-related cytokines ($IFN-{\gamma}$ and $TNF-{\alpha}$) in the AdH4-MVAH4 heterologous prime-boost regimen immunization group was significantly higher than that in the other viral vector-based vaccine- and BCG-immunized groups, respectively. These results indicate that an immunization regimen involving AdH4 may have a higher capacity to induce humoral and cellular immune responses against TB in mice than that by regimens containing BCG or MVAH4 alone, and the AdH4-MVAH4 prime-boost regimen may generate an ideal protective effect.

• Journal of Ginseng Research
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• 제41권4호
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• pp.548-555
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• 2017

Background: Korean Red Ginseng has been used for several decades to treat many diseases, enhancing both immunity and physical strength. Previous studies have documented the therapeutic effects of ginseng, including its anticancer, antiaging, and anti-inflammatory activities. These activities are mediated by ginsenosides present in the ginseng plant. Ginsenoside Rg3, an effective compound from red ginseng, has been shown to have antiplatelet activity in addition to its anticancer and anti-inflammatory activities. Platelets are important for both primary hemostasis and the repair of the vessels after injury; however, they also play a crucial role in the development of acute coronary diseases. We prepared ginsenoside Rg3-enriched red ginseng extract (Rg3-RGE) to examine its role in platelet physiology. Methods: To examine the effect of Rg3-RGE on platelet activation in vitro, platelet aggregation, granule secretion, intracellular calcium ($[Ca^{2+}]_i$) mobilization, flow cytometry, and immunoblot analysis were carried out using rat platelets. To examine the effect of Rg3-RGE on platelet activation in vivo, a collagen plus epinephrine-induced acute pulmonary thromboembolism mouse model was used. Results: We found that Rg3-RGE significantly inhibited collagen-induced platelet aggregation and $[Ca^{2+}]_i$ mobilization in a dose-dependent manner in addition to reducing ATP release from collagen-stimulated platelets. Furthermore, using immunoblot analysis, we found that Rg3-RGE markedly suppressed mitogen-activated protein kinase phosphorylation (i.e., extracellular stimuli-responsive kinase, Jun N-terminal kinase, p38) as well as the PI3K (phosphatidylinositol 3 kinase)/Akt pathway. Moreover, Rg3-RGE effectively reduced collagen plus epinephrine-induced mortality in mice. Conclusion: These data suggest that ginsenoside Rg3-RGE could be potentially be used as an antiplatelet therapeutic agent against platelet-mediated cardiovascular disorders.

• 약학회지
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• 제49권6호
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• pp.518-526
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• 2005

Hwanggumgung (HGG) is a hair-care product which is composed of several plant extracts used in oriental medicine. This study was carried out to investigate effect of HGG on hair regrowth in a shaving model of C57BL6 mice. Five-week-old mice were acclimated for 1 week under 23$\pm$3$^{\circ}C$, 50$\pm10\%$ relative humidity and 12 h of a light/dark cycle before beginning experiment. There were four experimental groups including distilled water (D.W., control), 10$\%$ ethanol (EtOH, vehicle control), a positive control of 3$\%$ minoxidil (MXD), and HGG for female and male mice, respectively; Six-weeks old mice were trimmed by electric clippers so as not to damage the skin. The next day; mice without visible scraches were selected, randomized and separated in groups of 11 mice. The test compounds were topically treated with 0.15ml per mouse per day for 21 days. The hair regrowth was photographically and histologically determined during the experimental period of 21 days. Enzyme activities of $\gamma$-glutamyl transpeptidase and alkaline phosphatase were also determined using a rate assay method. There were no clinical signs in all experimental groups. The topical application of 3$\%$ MXD and HGG in female mice promoted hair regrowth earlier and faster than the control groups. In male mice, the topical application of 3$\%$ MXD and HGG also accelerated hair growth compared with the controls. Ten percent ethanol also promoted hair growth faster than D.W group. The histology of hair growth in experimental groups was strongly associated with the hair regrowth. 3$\%$ MXD and HGG promoted elongation of hair follicles compared with the controls in both female and male mice. Activities of alkaline phosphatase and $\gamma$-glutamyl transpeptidase, enzymes related to hair growth, significantly increased after treatments of 3$\%$ MXD and HGG for 2 weeks in both female and male mice (p < 0.05). These results suggest that HGG has hair growth promoting activities and it can be for treatment for alopecia.

• IMMUNE NETWORK
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• 제11권1호
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• pp.79-94
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• 2011

Background: Dendritic cell (DC)-based vaccines are currently being evaluated as a novel strategy for tumor vaccination and immunotherapy. However, inducing long-term regression in established tumor-implanted mice is difficult. Here, we show that deoxypohophyllotoxin (DPT) induces maturation and activation of bone marrow-derived DCs via Toll-like receptor (TLR) 4 activation of MAPK and NF-${\kappa}B$. Methods: The phenotypic and functional maturation of DPT-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. DPT-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity and for tumor regression against melanoma. Results: DPT promoted the activation of $CD8^+$ T cells and the Th1 immune response by inducing IL-12 production in DCs. In a B16F10 melanoma-implanted mouse model, we demonstrated that DPT-treated DCs (DPT-DCs) enhance immune priming and regression of an established tumor in vivo. Furthermore, migration of DPT-DCs to the draining lymph nodes was induced via CCR7 upregulation. Mice that received DPT-DCs displayed enhanced antitumor therapeutic efficacy, which was associated with increased IFN-${\gamma}$ production and induction of cytotoxic T lymphocyte activity. Conclusion: These findings strongly suggest that the adjuvant effect of DPT in DC vaccination is associated with the polarization of T effector cells toward a Th1 phenotype and provides a potential therapeutic antitumor immunity.

• Journal of Ginseng Research
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• 제39권4호
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• pp.398-405
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• 2015

Background: Ginseng has been used as a tonic for invigoration of the human body. In a previous report, we identified a novel candidate responsible for the tonic role of ginseng, designated gintonin. Gintonin induces $[Ca^{2+}]_i$ transient in animal cells via lysophosphatidic acid receptor activation. Gintonin-mediated $[Ca^{2+}]_i$ transient is linked to anti-Alzheimer's activity in transgenic Alzheimer's disease animal model. The previous method for gintonin preparation included multiple steps. The aim of this study is to develop a simple method of gintonin fraction with a high yield. Methods: We developed a brief method to obtain gintonin using ethanol and water. We extracted ginseng with fermentation ethanol and fractionated the extract with water to obtain water-soluble and water-insoluble fractions. The water-insoluble precipitate, rather than the water-soluble supernatant, induced a large $[Ca^{2+}]_i$ transient in primary astrocytes. We designated this fraction as gintonin-enriched fraction (GEF). Results: The yield of GEF was approximately 6-fold higher than that obtained in the previous gintonin preparation method. The apparent molecular weight of GEF, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was equivalent to that obtained in the previous gintonin preparation method. GEF induced $[Ca^{2+}]_i$ transient in cortical astrocytes. The effective dose (ED50) was $0.3{\pm}0.09{\mu}g/mL$. GEF used the same signal transduction pathway as gintonin during $[Ca^{2+}]_i$ transient induction in mouse cortical astrocytes. Conclusion: Because GEF can be prepared through water precipitation of ginseng ethanol extract and is easily reproducible with high yield, it could be commercially utilized for the development of gintoninderived functional health food and natural medicine.

• Journal of Periodontal and Implant Science
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• 제37권4호
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• pp.859-869
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• 2007

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

• 한방비만학회지
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• 제15권1호
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• pp.9-23
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• 2015

Objectives: This study was investigated the improvement effects of Pakistani Ephedra herba-containing Gangji-hwan-1, 2, 3 (Di-fatty; DF-1, 2, 3), Chinese Ephedra herba-containing Gangjihwan-4 (DF-4) and combination of DF-1 and Gamisoche-hwan (GSH) on obesity in a high fat diet-fed obese mouse model. Methods: Eight-week-old C57BL/6N mice were divided into seven groups: a normal lean group given a standard diet, an obese control group given a high fat diet, and DF-1, 2, 3, 4, and DF-1+GSH groups given a high fat diet with DF-1, 2, 3, 4 (40, 80, 160, 80 mg/kg), and DF-1+ GSH (80 mg/kg), respectively. After 8 weeks of treatment, body weight gain, feeding efficiency ratio (FER), blood lipid markers, liver histology, and fat weight and histology were examined. Results: Body weight gain was significantly decreased in DF and DF-1+GSH groups compared with control. The extent of decreases was eminent in DF-1+GSH group. FER and circulating concentration of leptin were decreased in DF and DF-1+GSH groups compared with control. Circulating concentrations of triglyceride, glucose and insulin were decreased in DF and DF-1+GSH groups compared with control. The size of adipocytes were decreased by DF and DF-1+GSH groups compared with control, whereas the adipocyte number per unit area was increased by them, suggesting that DF and DF-1+GSH groups decreased the number of large adipocytes. Conclusions: In conclusion, these results suggest that DF and DF-1+GSH groups decrease FER, plasma leptin concentration, blood anti-obesity biomarkers and fat mass, improves body weight gain. In addition, these effects were more effective in DF-1+GSH combination group than in DF-1, 2, 3, 4 groups.

• 대한한방부인과학회지
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• 제18권3호
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• pp.110-126
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• 2005

Purpose : The Purpose of this research was to investigate the effects of antithrombotic activities of Wuslsan (WSS). Methods : Measure the effect which was given to blood flow rate through the regular volume of glass tube after the blood was diluted five times with ACD soulution. Antithrombotic effect was calculated as a percentage of the experimental animal figure protected from the paralysis of hind legs or death of the mouse that is caused from the administration of platelet aggregation regent. Being classified one group of eight mice, each of them was divided into Normal, Control, and WSS. The normal group supplied a saline solution and the control group brought the dextran extravasated blood after an hour of administering the saline solution. Also WSS was dissolved in 2ml saline solution and then we dosed it to the experimental mice with Oral Zonde one day before the experiment. After that, the mice were abstained from food. And then we gave a measured amount of it before an hour. Finally, it gave rise to dextran extravasated blood in the same way as the Control group. Results : The results were obtained as follows. WSS inhibited platelet aggregation induced by ADP and epinephrine significantly as compared with the control group. WSS showed fibrinolytic activity insignificantly as compared with the control group. WSS increased blood flow rate significantly as compared with the control group in vitro. WSS inhibited pulmonary embolism induced by collagen and Epinephrine(inhibitive rate is 37.5%). WSS increased number of platelet and fibrinogen amount significantly, and shortened PT and APTT as compared with the control group in thrombus model induced by dextran. Conclusion : WSS is effective antithrombotic activity from experimental result.

• 대한한방부인과학회지
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• 제31권1호
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• pp.20-42
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• 2018

Objectives: This study was designed to investigate anti-obesity effects the improvement effects of Gyejibongnyeong-hwan and Gyejibongnyeong-hwan-Gangji-hwan (CIPPDF) in a ob/ob mouse model. Methods: Seven-week old mice (wild-type C57BL/6J and ob/ob) were used for all experiments. Wild-type C57BL/6J mice were used as normal group and obese ob/ob mice were randomly divided into 4 groups. a normal group given a standard diet, an obese control group given a standard diet with CIPP (300 mg/kg), CIPPDF (1) (300+300 mg/kg), CIPPDF (2) (300+600 mg/kg) respectively. After 10 weeks of treatment, body weight gain, feeding efficiency ratio, blood lipid markers, mRNA levels of genes involved in fatty acid ${\beta}-oxidation$ and lipogenesis in in-vivo, were examined. Results: 1. Body weight gain and Feeding efficiency ratio were significantly decreased in CIPPDF (1) compared with control. Fat mass was significantly decreased in CIPPDF (2) in EAT compared with control. 2. Consistent their effects on body weight gain and fat mass, circulating concentrations of LDL-cholesterol were decreased in CIPPDF (1), CIPPDF (2) groups compared with control. 3. MCAD mRNA levels of genes was increased in CIPPDF (1), CIPPDF (2) groups in the liver, epididymal adipose tissue compared with control. VLCAD mRNA levels of genes was increased in CIPPDF (1), CIPPDF (2) groups in the skeletal muscle compared with control. 4. $PPAR{\gamma}$ mRNA was decreased in CIPPDF (1) in the liver compared with control. SCD1 mRNA was decreased in CIPPDF (1), CIPPDF (2) groups in the epididymal adipose tissue compared with control. Conclusions: In conclusion, These results suggest that CIPPDF not only decrease feeding efficiency ratio, and LDL-cholesterol, but also reduce EAT fat mass contributing to the improvement of ovesity. CIPPDF also were increased in mRNA levels of genes involved in fatty acid ${\beta}-oxidation$ and decreased in mRNA levels of genes involved in lipogenesis.