• 대한화장품학회지
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• 제32권3호
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• pp.201-208
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• 2006

본 연구는 예로부터 민간요법 및 한방에 유용한 약재로 사용되어지고 있는 관중(Dryopteris crassirhizoma Nakai)을 이용하여 Propionibacterium acnes에 대한 항균활성을 평가하고, 이를 토대로 여드름 억제 및 치료에 관련된 화장품 소재를 개발하는데 그 목적이 있다. P. acnes에 대한 관중 crude 추출물 및 헥산분획물의 minimum inhibitory concentration (MIC) 및 paper disk diffusion assay로 항균 효과를 측정하였으며, 추출물에 대한 온도 및 pH 변화에 따른 안정성을 평가하였다 추출물을 이용한 P. acnes에 대한 MIC 값은 관중 crude 추출물은 0.008 mg/mL, 헥산추출물은 0.001 mg/mL로 측정되었으며, 이는 대조군으로 사용한 항생제인 triclosan의 MIC 값(0.004 mg/mL)과 비교했을 때 시료가 관중 crude 추출물 및 헥산 분획물임을 고려하면 매우 항균활성이 높은 것으로 평가되었다. 추출물에 대한 paper disk diffusion assay를 수행한 결과, 대조군으로 사용한 triclosan과 대비, 높은 활성을 나타내는 것으로 측정되었으며, HaCaT 세포에 대한 독성은 $20{\mu}g/mL$ 이하의 농도에서는 나타나지 않았다. 관중추출물의 온도, pH 안정성을 평가한 결과, 25, 70, 80, 90, 100, $121^{\circ}C$ 등 다양한 온도 조건과 $pH 2{\sim}pH 11$의 변화에 있어서도 그 효능이 변하지 않고 안정된 상태로 유지되는 것으로 관찰되었다.

• Restorative Dentistry and Endodontics
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• 제34권3호
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• pp.192-198
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• 2009

이번 연구는 치근 천공의 치료 재료인 white mineral trioxide aggregate (MTA)를 흔히 사용되는 calcium hydroxide liner ($Dycal^{(R)}$), glass ionomer cement (GIC), 그리고 MTA와 유사한 성분을 가진 Portland cement와 세포독성 실험으로 생체 친화성을 평가하는 것이다. 세포독성의 정도는 MG-63 세포를 이용해 주사전자 현미경적 관찰과 수용성 tetrazolium salt를 이용한 흡광도를 측정 (XTT assay)하여 평가하였다. SEM 관찰에서, 1일과 3일째 모두에서 GIC와 MTA, Portland cement 표면에서는 잘 부착된 세포를 보여주었다. 반면에, Dycal 표면의 세포들은 둥글고 부착되지 않은 양상을 보여 주었다. XTT assay에서는 Dycal을 제외한 모든 재료에서 유사하게 높은 세포 활성도를 보여주었으며, 이는 SEM 관찰 소견과 일치하였다. 이번 연구는 MTA가 생체친화적인 재료라는 견해를 뒷받침한다. 또한 Portland cement와 GIC에서도 MTA와 유사한 세포반응을 보여주었다.

• 한국가축번식학회지
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• 제9권2호
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• pp.133-139
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• 1985

An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.

• 대한한방내과학회지
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• 제32권2호
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• pp.153-169
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• 2011

Objectives : This study was performed to investigate the anti-fibrogenic effect and the effect on cell growth and apoptosis in YJCGT, YJCGT YSO and YJCGT YSCO on thioacetamide-induced rat liver tissue and the immortalized human hepatic cell line LX2. Materials and Methods : LX2 cells were treated with various concentrations (0, 50, 150, 300 ug/ml) of YJCGT, Y+YSO, and Y+YSCO extract for 24, 48 and 72 hours. After the treatment, cell viability was measured by using MTT assay. Caspase inhibitor assay, and cell viability were determined by a colorimetric assay with PMS/MTS solution. Rat liver fibrosis was induced by intraperitoneal thioacetamide injection 150 mg/kg 3 times a week for 5 weeks. After the treatment, body weight, liver & spleen weights, liver function test, the complete blood cell count and the change of portal pressure were studied. After YJCGT, Y+YSO, and Y+YSCO treatment, percentages of collagen in thioacetamide-induced rat liver tissue were measured. Results : The viability of the LX2 cell decreased in a dose- and time-dependent manner. Exposure of LX2 cells to YJCGT, YJCGT+YSO and YJCGT+YSCO induced caspase-3 activation, but co-treatment of YJCGT, YJCGT+YSO and YJCGT+YSCO with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. There was no difference in rat body weight between the thioacetamide only group and the YJCGT, YJCGT+YSO and YJCGT+YSCO groups. In the YJCGT, YJCGT YSO and YJCGT YSCO groups, the serum level of GPT significantly went down compared with the thioacetamide only group. In the YJCGT, Y+YSO, Y+YSCO groups, white blood cell elevated by thioacetamide injection decreased but RBC, Hgb, and Hct increased. In the Y+YSO group, the portal pressure elevated by thioacetamide injection significantly decreased. In the histological finding, thioacetamide injections caused severe fibrosis, but YJCGT, Y+YSO, and Y+YSCO treatment significantly reduced the amounts of hepatic collagens. Conclusions : YJCGT, Y+YSO, and Y+YSCO inhibit the growth of LX2 cells by inducing apoptosis through caspase activity. YJCGT, Y+YSO, and Y+YSCO have beneficial effects on the treatment of cirrhotic patients as well as patients with chronic hepatitis.

• 대한한방내과학회지
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• 제18권1호
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• pp.231-241
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• 1997

국내 사망률중 가장 높은 비율을 차지하고 있는 악성종양(惡性腫瘍)에 대하여 국내외적으로 다양하게 연구되고 있다. 그러나 아직까지도 종양(腫瘍)을 치료하기 우하여 수술요법(手術療法) 방사선요법(放射線療法) 면역요법(免疫療法) 화학요법(化學療法) 등의 많은 치료법들을 개발하고 있지만 종양(腫瘍)에 대한 치료원칙이나 치료약물에 대하여서는 아직까지 미흡한 것이 국내의 현실이다. 현재 일반적으로 항암제(抗癌劑)를 이용한 화학요법(化學療法) 등이 사용되고 있지만 이에 따른 부작용(副作用)이 많아 항암제(抗癌劑)와 한약재(韓藥材)를 병용투여(倂用投與)함으로써 부작용(副作用)을 최소화하고, 이에 따라 한약재(韓藥材)가 정상세포(正常細胞)에 영향을 미치며, 암종세포(癌腫細胞)에는 영향을 미칠 것으로 사료되어 관찰한 결과 유의성(有意性)이 있어 보고하게 되었다. 그리하여 수종(數種)의 한약재중(韓藥材中) 청열작용(淸熱作用)과 활혈화어(活血化瘀)작용이 있는 대극(大戟)과 목단피(牧丹皮)를 인체의 피부암세포(皮膚癌細胞)인 A431 세포(細胞), 자궁암세포(子宮癌細胞)인 HeLa 세포(細胞), 급성백혈병세포(急性白血病細胞)인 MOLT-4 세포(細胞), 만성골수성백혈병세포(慢性骨髓性白血病細胞)인 K562 세포(細胞)에 대한 세포독성(細胞毒性)과 항암제(抗癌劑)인 mitomycin C와 병용(倂用)처리결과를 MTT assay를 통하여 관찰하였다. 또한 정상세포(正常細胞)에 대한 세포독성(細胞毒性)을 검색하기 위하여 마우스 섬유아세포(Balb/c 3T3), 마우스 흉선(胸腺) 및 비장세포(脾臟細胞), human lymphocyte에 미치는 세포독성(細胞毒性)을 검토하였다. 대극(大戟)과 목단피(牧丹皮)는 A431 세포(細胞)와 K562 세포(細胞), 마우스 섬유아세포(細胞)인 Balb/c 3T3 세포(細胞) 및 mitomycin C와 병용처리(倂用處理)하였을 때 mitomycin C를 단독처리하였을 때보다 A431 세포(細胞)의 증식을 억제하였고, 백선피(白蘚皮)와 천산갑(穿山甲)은 human lymphocyte의 증식을 촉진하였다.

• Tuberculosis and Respiratory Diseases
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• 제71권2호
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• pp.88-96
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• 2011

Background: It is known that cigarette smoke (CS) causes cell death. Apoptotic cell death is involved in the pathogenesis of CS-related lung diseases. Some members of the protein kinase C (PKC) family have roles in cigarette smoke extract (CSE)-induced apoptosis. This study was conducted to investigate the role of PKC epsilon in CSE-induced apoptosis in human lung fibroblast cell line, MRC-5. Methods: Lactate dehydrogenase release was measured using a cytotoxicity detection kit. The MTT assay was used to measure cell viability. Western immunoblot, Hoechst 33342 staining and flow cytometry were used to demonstrate the effect of $PKC{\varepsilon}$. Caspase-3 and caspase-8 activities were determined using a colorimetric assay. To examine $PKC{\varepsilon}$ activation, Western blotting was performed using both fractions of membrane and cytosol. Results: We showed that CSE activated $PKC{\varepsilon}$ by demonstrating increased expression of $PKC{\varepsilon}$ in the plasma membrane fraction. Pre-treatment of $PKC{\varepsilon}$ peptide inhibitor attenuated CSE-induced apoptotic cell death, as demonstrated by the MTT assay (13.03% of control, 85.66% of CSE-treatment, and 53.73% of $PKC{\varepsilon}$ peptide inhibitor-pre-treatment, respectively), Hoechst 33342 staining, and flow cytometry (85.64% of CSE-treatment, 53.73% of $PKC{\varepsilon}$ peptide inhibitor-pre-treatment). Pre-treatment of $PKC{\varepsilon}$ peptide inhibitor reduced caspase-3 expression and attenuated caspase-3, caspase-8 activity compared with CSE treatment alone. Conclusion: $PKC{\varepsilon}$ seem to have pro-apoptotic function and exerts its function through the extrinsic apoptotic pathway in CSE-exposed MRC-5 cells. This study suggests that $PKC{\varepsilon}$ inhibition may be a therapeutic strategy in CS-related lung disease such as chronic obstructive pulmonary disease.

• 대한본초학회지
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• 제26권4호
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• pp.39-47
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• 2011

Objective: The roots, leaves, flowers, stems and seeds of Cirsium japonicum var. ussuriense are often used in treatment of human diseases such as hemorrhage, blood congestion and inflammation. Focusing our attention on natural and bioavailable sources of antioxidants and anti-inflammation, we undertook to investigate the antioxidant and anti-inflammatory properties of Cirsium japonicum var. ussuriense used as a folk medicine in Korea. Methods: The extracts of the leaves, stems, flowers, seeds and roots from C. japonicum var. ussuriense were prepared by extracting with water or 80% ethanol. Total flavonoids and polyphenols were measured by a colorimetric assay. The free radical scavenging activity of the extract was analyzed by the DPPH (1,1-diphenyl-2-picryl hydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) and Griess reagent assay. An oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The level of prostaglandin $E_2$ ($PGE_2$) was measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blot analysis. Results: Total flavonoid and polyphenol amounts of the leaves (CLE) and flowers (CFE) showed higher than those of the seed extract (CSE), stem extract (CSTE) and roots (CRE). CLE and CFE also showed the high antioxidant activities such as DPPH, NO-like and ABTS radical scavenging activity. An antioxidant activities of these water extracts showed higher than those of 80% ethanol extracts. We investigated the anti-inflammatory effects of CLE on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. CLE significantly suppressed the levels of the inflammatory mediators such as NO and prostaglandin $E_2$ ($PGE_2$) in dose dependant. Furthermore, the levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with CLE extract in a dose dependent manner. Conclusions: These results suggest that CLE water extract has a higher anoxidant and anti-inflammatory activity, these properties may contribute to the oxidative and inflammatory related disease care.

• Nutrition Research and Practice
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• 제16권6호
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• pp.685-699
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• 2022

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

• 대한본초학회지
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• 제28권1호
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• pp.59-64
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• 2013

Objectives : Methicillin-Resistant Staphylococcus aureus (MRSA) is a cephalosporin and beta-lactam antibiotic-resistant strains. In most cases, that is spread from infected patients and infection rates are growing increasingly. Thus, accordingly, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infections diseases. Methods : The antibacterial activities of Sinhyowoldosan were evaluated against 3 strains of Methicillin-resistant staphylococcus aureus(MRSA) and 1 standard Methicillin-susceptible S. aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay was performed under dark. Results : The MIC (minimum inhibitory concentration) of Sinhyowoldosan water extract against S. aureus strains ranged from 500 to 2,000 ${\mu}g/mL$, so we have confirmed it on a strong antibacterial effect. Also, the combinations of Sinhyowoldosan water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Sinhyowoldosan water extract against MRSA have antibacterial activity, it has potential as alternatives to antibiotic agent. the combination test was used, Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of SH is affected by the cell membrane were assessed. Conclusion : We suggest that the Sinhyowoldosan water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.

• Asian Pacific Journal of Cancer Prevention
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• 제16권5호
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• pp.1771-1779
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• 2015

Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide. Presently, targeted therapy via monoclonal antibodies to specific tumor-associated antigens is being continuously developed. Hep88 mAb has proven to exert tumoricidal effects on the HepG2 cell via a paraptosis-like morphology. To verify the pathway, we then demonstrated downstream up-regulation of caspase-3, caspase-8 and caspase-9, assessingmRNA expression by real-time PCR and associated enzyme activity by colorimetric assay. Active caspase-3 determination was also accomplished by flow cytometry. Active caspase-3 expression was increased by Hep88 mAb treatment in a dose-and time-dependent manner. All of the results indicated that Hep88 mAb induced programmed cell death in the HepG2 cell line from paraptosis-like to apoptosis by downstream induction of caspases. These conclusions imply that Hep88mAb might be a promising tool for the effective treatment of HCC in the future.