• 제목/요약/키워드: MTT method.

검색결과 423건 처리시간 0.041초

발효 애엽(艾葉) 추출물이 인간 간암세포주 HepG2 활성에 미치는 영향 (Effect of Fermented Artemisiae Argyi Folium on Human Hepatoma Cell Line HepG2 Activity)

  • 한효상
    • 대한본초학회지
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    • 제28권3호
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    • pp.107-113
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    • 2013
  • Objective : The purpose of this study was to investigate the effect of fermented Artemisiae Argyi Folium(AAF) on some activities of human hepatoma cell, HepG2. Method : To investigate the effect of fermented Artemisiae Argyi Folium(AAF) activity on the human hepatoma cells, AAF extracts was fermented by Lactobacillus pentosus K34(AFL) and Sacchromyces cerevisiae STV89(AFS). And the effects of AFL or AFS on the activities of HepG2 cell, such as cell viability, nitric oxide(NO) production and reactive oxygen species(ROS) production, were tested. Result : Human Hepatoma Cells were incubated each for 3 hours and 24 hours. Human Hepatoma Cells treated with the extract was measured with MTT assay. Then AFL was found to be non-toxic at concentrations of 10 ug/mL(3h), 100 ug/mL(24h) or more. AFS was the same result at concentrations of more than 10 ug/mL. The extract increased ROS generation in Human Hepatoma Cells. AFL increased at concentrations of 100 ug/mL more (3h, also 10 ug/mL more) and 50 ug/mL(24h) and AFS increased both 50 ug/mL. In point of NO generation, AFL inhibited at concentrations of 10 ug/mL(3h) and 100 ug/mL(24h) more (3h, also 10 ug/mL more) and AFS also inhibited 50 ug/mL or more. Conclusion : AFL and AFS, obtained from Artemisiae Argyi Folium extracts by fermentation, reduced the NO production and increased ROS production in HepG2 cell, without cytotoxicity on HepG2 cell. The results suggested that AFL and AFS increased the immunological effects of Artemisiae Argyi Folium extracts.

Protective effect and mechanism of ginsenoside Rg2 on atherosclerosis

  • Qianqian Xue;Tao Yu;Zhibin Wang;Xiuxiu Fu;Xiaoxin Li;Lu Zou;Min Li;Jae Youl Cho;Yanyan Yang
    • Journal of Ginseng Research
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    • 제47권2호
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    • pp.237-245
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    • 2023
  • Background: Ginsenoside Rg2 (Rg2) has a variety of pharmacological activities and provides benefits during inflammation, cancer, and other diseases. However, there are no reports about the relationship between Rg2 and atherosclerosis. Methods: We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to detect the cell viability of Rg2 in vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs). The expression of inflammatory factors in HUVECs and the expression of phenotypic transformation-related marker in VSMCs were detected at mRNA levels. Western blot method was used to detect the expression of inflammation pathways and the expression of phenotypic transformation at the protein levels. The rat carotid balloon injury model was performed to explore the effect of Rg2 on inflammation and phenotypic transformation in vivo. Results: Rg2 decreased the expression of inflammatory factors induced by lipopolysaccharide in HUVECs-without affecting cell viability. These events depend on the blocking regulation of NF-κB and p-ERK signaling pathway. In VSMCs, Rg2 can inhibit the proliferation, migration, and phenotypic transformation of VSMCs induced by platelet derived growth factor-BB (PDGF-BB)-which may contribute to its anti-atherosclerotic role. In rats with carotid balloon injury, Rg2 can reduce intimal proliferation after injury, regulate the inflammatory pathway to reduce inflammatory response, and also suppress the phenotypic transformation of VSMCs. Conclusion: These results suggest that Rg2 can exert its anti-atherosclerotic effect at the cellular level and animal level, which provides a more sufficient basis for ginseng as a functional dietary regulator.

3T3-L1 지방 전구세포의 분화 및 지방 생성에 미치는 Fragaria Orientalis L. 물 추출물의 영향 (Effects of Fragaria Orientalis Water extract on Adipogenesis and Cell Differentiation in 3T3-L1 Cells)

  • 최문열;김미형;김미려
    • 대한본초학회지
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    • 제39권3호
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    • pp.49-56
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    • 2024
  • Objective : Obesity, which has recently been rapidly increasing in the obese population, is caused by an imbalance in energy intake and consumption. The reason why we need to manage obesity well is that the prevalence of complications such as diabetes, atherosclerosis, insulin resistance, and cardiovascular disease increases. In this study, the effect of FO (Fragaria orientalis) water extract on fat metabolism in 3T3-L1 cells was observed to develop a new anti-obesity material based on Mongolian medical books. Methods : The effect of FO extract on adipogenesis in 3T3-L1 cells was observed using DPPH scavenging, pancreatic lipase inhibitory activity, MTT analysis and Oil-red-O staining method. And the expression of proteins related to lipid metabolism was analyzed by Western blot. Results : The FO group significantly increased the DPPH radical scavenging activity at 5 mg/ml compared to the positive control BHA at 0.1 mg/ml. In oil red O staining at a safe concentration without cytotoxicity, lipid accumulation was significantly inhibited by less than 80% compared to the control group at all concentrations. Moreover, treatment of FO significantly increased the expression of proteins related to lipid metabolism, such as p-AMPK and p-ACC, in 3T3-L1 cells, and the expression of CPT-1 tended to increase in a dose-dependent manner. However, the expression of PPAR-γ was significantly decreased in a dose-dependent manner. Conclusion : These results suggest that FO water extract has a potential anti-obesity effect and are expected to be utilized in the development of materials for obesity prevention and treatment.

생쥐 대장 카할세포에서 가미소요산, 반하사심탕 및 보중익기탕의 효과에 관한 비교연구 (Effects of Gamisoyo-san, Banhasasim-tang and Bojungikki-tang in Colonic Interstitial cells of Cajal in mice)

  • 최나리;최우균;김병주
    • 대한한의학방제학회지
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    • 제32권1호
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    • pp.29-37
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    • 2024
  • Objectives : The purpose of this study was to examine the effects of insurance herbal medicines on colonic interstitial Cells of Cajal (ICC) in mice. Methods : Among the insurance herbal medicines, we chose Gamisoyo-san (GSS), Banhasasim-tang (BHSST) and Bojungikki-tang (BGIKT). We made the ICC culture in large intestine in mice and used the electrophysiological method to record pacemaker potentials. Also we used MTT assay to check cell viability and examined the ICC protein expression by western blot. Results : 1. GSS (1-10 mg/ml) induced the pacemaker potential depolarization and decreased frequency with concentration-dependent manners in colonic ICC. EC50 is 2.99 mg/ml. BHSST (1-10 mg/ml) induced the pacemaker potential depolarization and decreased frequency with concentration-dependent manners in colonic ICC. EC50 is 2.76 mg/ml. BGIKT (1-10 mg/ml) induced the pacemaker potential depolarization and decreased frequency with concentration-dependent manners in colonic ICC. EC50 is 4.49 mg/ml. 2. GSS, BHSST and BGIKT had no effects on cell viability in colonic ICC. 3. GSS and BGIKT increased the Anoctamin-1 (ANO1) protein expression and BHSST increased the transient receptor potential melastatin-subfamily member 7 (TRPM7) protein expression in colonic ICC. Conclusions : These results suggest that GSS, BHSST, and BGIKT have shown the potential to regulate gastrointestinal (GI) motility by regulating colonic ICC and may show the potential to treat colon-derived GI diseases such as irritable bowel syndrome (IBS).

우각승마탕이 LPS로 유도된 RAW 264.7 세포에서 NO 생산에 미치는 영향 (The Effect of Woogakseungmatang Extract on NO Production in LPS- Stimulated RAW 264.7 Cells)

  • 조나영
    • Korean Journal of Acupuncture
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    • 제35권4호
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    • pp.166-173
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    • 2018
  • 목적 : 우각승마탕은 한의학에서 안면마비 등 안면부의 통증 및 마비를 동반하는 질환에 주로 사용하는 처방이다. 본 연구의 목적은 LPS로 염증 반응을 유도한 대식세포에서 우각승마탕의 항염증 및 항산화 효과를 알아 보는 것이다. 방법 : 동의보감 처방에 기반한 우각승마탕을 열수 추출한 후 동결건조 하였다. MTT 방법을 이용하여 세포 독성을 평가하였고, 우각승마탕 추출물을 처리한 RAW 세포에서 NO 생산 및 $PGE_2$ 생성량을 조사 하였다. 또한 염증과 관련된 사이토카인($IL-1{\beta}$, IL-6, and $TNF-{\alpha}$)의 생성량 변화를 조사 하였다. 우각승마탕의 항산화 효능은 DPPH 법으로 측정 하였다. 결과 : 우각승마탕 추출물을 처리 한 대식세포는 $100{\mu}g/ml$ 농도에서 $92.18{\pm}2.97%$의 세포 생존율을 보였다. 150과 $200{\mu}g/ml$ 농도에서 세포생존율은 각각 $85.39{\pm}1.54$ and $76.11{\pm}2.81%$로 통계적으로 유의한 감소가 있었다. $100{\mu}g/ml$ 농도에서 NO 생성량은 $85.13{\pm}1.93%$ 였으며, 대조군과 비교하여 유의한 감소가 나타났다. $PGE_2$ 생성량은 $100{\mu}g/ml$ 농도에서 $82.64{\pm}2.54%$ 이었고, 통계적으로 유의한 감소가 있었다. $IL-1{\beta}$, IL-6, $TNF-{\alpha}$$100{\mu}g/ml$에서 각각 $85.24{\pm}1.18$, $86.18{\pm}3.37$, $84.20{\pm}3.94%$였으며, 대조군과 비교하여 통계적으로 유의한 감소를 나타내었다. DPPH free radical 소거능은 $100{\mu}g/ml$에서 $50.89{\pm}3.59%$를 나타내었다. 결론 : 본 연구의 결과로 미루어 볼 때 우각승마탕은 세포독성을 보이지 않는 최고 농도인 $100{\mu}g/ml$에서 미약한 항염증 효과를 나타내었고 농도의존적 항염증 효과를 확인되지 않았다. 따라서 본 연구는 우각승마탕의 잠재적 항염증 효과를 확인하는 의미가 있었다. 이 연구를 바탕으로 우각승마탕의 안전하고 효과적인 임상적 사용을 위하여 체계적이고 다양한 연구가 수행되어야 할 것이다.

밀리미터파 대역 차량용 레이더를 위한 순서통계 기법을 이용한 다중표적의 데이터 연관 필터 (Multi-target Data Association Filter Based on Order Statistics for Millimeter-wave Automotive Radar)

  • 이문식;김용훈
    • 대한전자공학회논문지SP
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    • 제37권5호
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    • pp.94-104
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    • 2000
  • 차량 충돌 경보용 레이더 시스템의 개발에 있어 표적 추적의 정확도와 신뢰도는 매우 중요한 요소이다. 여러 표적을 동시에 추적할 때 중요한 것은 표적과 측정치와의 데이터 연관(data association) 이며, 부적절한 측정치가 어느 표적과 연관되면 그 표적은 트랙을 벗어나 추적능력을 잃어버릴 수 있고 심지어 다른 표적의 추적에도 영향을 줄 수 있다 지금까지 발표된 대부분의 데이터 연관 필터들은 근접하여 이동하는 표적들의 경우 이와 같은 문제점을 보여왔다 따라서, 현재 개발되고 있는 많은 알고리즘들은 이러한 데이터 연 관 문제의 해결에 초점을 맞추고 있다 본 논문에서는 순서통계(order statistics)를 이용한 새로운 다중 표적의 데이터 연관 방법에 대하여 서술하고자 한다 OSPDA와 OSJPDA로 불리는 제안된 방법은 각각 PDA 필터 또는 JPDA 필터에서 계산된 연관 확률을 이용하며 이 연관 확률을 결정 논리(dicision logic)에 의한 가중치로 함수화 하여 표적과 측정치 사이에 최적 혹은 최적 근처의(near optimal) 데이터 연관이 가능하도록 한 것이다 시뮬레이션 결과를 통해, 제안한 방법은 기존의 NN 필터, PDA 필터, 그리고 JPDA 필터의 성능과 비교 분석되었으며, 그 결과 제안한 OSPDA, OSJPDA 필터는 PDA, JPDA 필터보다 추적 정확도에 대해 각각 약 18%, 19% 이상으로 성능이 향상됨을 확인하였다 제안한 방법은 CAN을 통해 차량 엔진 등의 ECU와 통신하도록 개발된 DSP 보드를 이용하여 구현되었다

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육계약침액이 LPS로 유도된 대식세포의 염증반응에 미치는 영향 (Effects of $Cinnamomi$ $Cortex$ Pharmacopuncture on LPS-induced Inflammatory Response in Macrophage)

  • 김형석;노정두
    • Journal of Acupuncture Research
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    • 제29권1호
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    • pp.15-24
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    • 2012
  • Objectives : In recent years, many studies have been widely researching anti-inflammation effect of various medicinal plants. $Cinnamomi$ $Cortex$ was not enough in researching of the anti-inflammation. Moreover, there is no comparative study about extraction methods. Therefore, we investigated the inhibitory effects of $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction on Nitric oxide(NO), Prostaglandin E2(PGE2) production, Cyclooxygenase(COX)-2, inducible NOS(iNOS) expression and extracellular signal regulate kinase(ERK)1/2 phosphorylation in lipopolysaccharide(LPS) induced RAW 264.7 macrophage cell. Methods : $Cinnamomi$ $Cortex$ was extracted by EtOH and Hot water. RAW 264.7 macrophage cell viability was measured by MTT assay. Effect of $Cinnamomi$ $Cortex$ pharmacopuncture on NO and PGE2 production in LPS induced macrophages was accessed by Griess assay and enzyme-linked immunospecific assay(ELISA), respectively. Inhibition effect on COX-2, iNOS expression and ERK1/2 phosphorylation was examined by Immunoblotting assay. Results : 1. Cytotoxic effect of $Cinnamomi$ $Cortex$ pharmacopuncture by Hot water extraction in RAW 264.7 macrophages was not appeared, except $3125{\mu}g/m{\ell}$. And cytotoxic effect was not appeared in EtOH extraction method. 2. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited NO production in LPS induced macrophages significantly. 3. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited PGE2 production in LPS induced macrophages significantly. 4. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited COX-2, iNOS expression in LPS induced macrophages. Especially, it has been confirmed that COX-2, iNOS expression were effectively inhibited in Hot water extraction. 5. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited ERK1/2 phosphorylation in LPS induced macrophages. Especially, it has been confirmed that ERK1/2 phosphorylation was effectively inhibited in Hot water extraction. Conclusions : According to the results, $Cinnamomi$ $Cortex$ pharmacopuncture suppresses NO, PGE2 production, COX-2, iNOS expression and ERK1/2 phosphorylation in LPS induced macrophages. It has a potential for treating various inflammatory diseases, and Hot water extraction method could be used more extensively than EtOH extraction method.

Radiosensitivity Enhancement by Arsenic Trioxide in Conjunction with Hyperthermia in the EC-1 Esophageal Carcinoma Cell Line

  • Cui, Yan-Hui;Liang, Hai-Jun;Zhang, Qing-Qin;Li, Si-Qing;Li, Xiao-Rui;Huo, Xiao-Qing;Yang, Qing-Hui;Li, Wei-Wei;Gu, Jian-Fa;Hua, Qin-Liang;Lu, Ping;Miao, Zhan-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권4호
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    • pp.1693-1697
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    • 2012
  • Objective: To explore the effect on radiosensitivity of arsenic trioxide ($As_20_3$) in conjunction with hyperthermia on the esophageal carcinoma EC-1 cell line. Method: Inhibition of EC-1 cell proliferation at different concentrations of $As_20_3$ was assessed using the methyl thiazolyl blue colorimetric method (MTT method), with calculation of $IC_{50}$ value and choice of 20% of the $IC_{50}$ as the experimental drug concentration. Blank control, $As_20_3$, hyperthermia, radiotherapy group, $As_20_3$ + hyperthermia, $As_20_3$ + radiotherapy, hyperthermia + radiotherapy and $As_20_3$ + hyperthermia + radiotherapy groups were established, and the cell survival fraction (SF) was calculated from flat panel colony forming analysis, and fitted by the 'multitarget click mathematical model'. Flow cytometry (FCM) was used to detect changes in cell apoptosis and the cell cycle. Results: $As_20_3$ exerted inhibitory effects on proliferation of esophageal carcinoma EC-1 cells, with an $IC_{50}$ of 18.7 ${\mu}mol/L$. After joint therapy of $As_20_3$ + hyperthermia + radiotherapy, the results of FCM showed that cells could be arrested in the $G_2$/M phase, and as the ratio of cells in $G_0/G_1$ and S phases decreased, cell death became more pronounced. Conclusion: $As_20_3$ and hyperthermia exert radiosensitivity effects on esophageal carcinoma EC-1 cells, with synergy in combination. Mechanistically, $As_20_3$ and hyperthermia mainly influence the cell cycle distribution of EC-1 esophageal carcinoma cells, decreasing the repair of sublethal damage and inducing apoptosis, thereby enhancing the killing effects of radioactive rays.

박막증착시 티타늄 표면의 마손저항도와 세포독성에 관한 연구 (A STUDY ON THE RESISTANCE OF WEAR AND CYTOTOXICITY OF THE TITANIUM SURFACE AFTER FILM DEPOSITIONS)

  • 김형우;김창회;김영수
    • 대한치과보철학회지
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    • 제39권1호
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    • pp.84-95
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    • 2001
  • Titanium is widely used in dentistry for its low density, high strength, fatigue resistance, corrosion resistance, and biocompatibility. But it has a tendency of surface damage under circumstance of friction and impact for its low hardness of the surface. Coating is one of methods fir increasing surface hardness. Its effect is to improve surface physical characteristics without change of titanium. Diamond-like carbon and titanium nitride are known for its high hardness of the surface. So that this study was aimed at the wear test and the cytotoxicity test of the commercially pure titanium and Ti-6Al-4V alloy which were deposited by diamond-like carbon film or titanium nitride film to acertain improvement of the surface hardness and the biocompatibility. A disk (25mm diameter, 2mm thickness) was made of commercially pure titanium and Ti-6Al-4V alloy and these substrates were deposited by diamond-like carbon film or titanium nitride film. Diamond-like carbon film was deposited by the method of radiofrequency plasma assisted chemical vapor deposition and titanium nitride film was deposited by the method of reactive arc ion plating. Then these substrates were tested about wear characteristics by the pin-on-disk type wear tester in which ruby ball was used as a wear causer under the load of 32N, The fracture cycles were measured by rotating the substrates until their films were fractured. The wear volume was measured after 150 cycles and 3,000 cycles using surface profiler. The cytotoxicity test was peformed by the method of the MTT assay. The results were as follows : 1. In the results of the wear volume test, commercially pure titanium and titanium alloy which were coated by diamond-like carbon film or titanium nitride aim had higher resistance against wear than the substrates which were not coated by any films (P<0.05). 2. In the results of the fracture cycle test and the wear volume test, diamond-like carbon film had higher resistance against wear than titanium nitride film (P<0.05). 3. In both coatings of diamond-like carbon aim and titanium nitride film, Ti-6Al-4V alloy had higher resistance against wear than commercially pure titanium (P<0.05) 4. In the results of the cytotoxicity test, diamond-like carbon film and titanium nitride film had little cytotoxicity as like commercially pure titanium or Ti-6Al-4V alloy (P>0.05).

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화침의 안전성 평가에 관한 고찰 (The Safety Assessment of Fire needling)

  • 연선희;이새봄;권오상;조성진;최광호;이상훈;최선미;류연희
    • 한국한의학연구원논문집
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    • 제18권3호
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    • pp.103-110
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    • 2012
  • Objectives : Fire needling has been applied as the treatment for various diseases and been getting much attention from Oriental medicine due to its excellent effectiveness as the results of clinical studies have reported. However, the research findings on the safety of treatment method, materials for the Fire needling needle materials and the possibility of burn injury during the procedure are still insufficient. Methods : A thermo imaging camera was used to confirm the temperature distribution on acupuncture needle and the treatment area during the fire needling therapy. Then the degree of thermal injury was observed by H&E stain and TUNEL assay. In addition, in order to assess the safety of acupuncture materials, we conducted MTT assay using a L6 cell line. Results : The average temperature of the skin surface was observed at $47{\sim}51^{\circ}C$ after classic fire needling and $30^{\circ}C$ after warming fire needling. Warming fire needling therapy does not induce a burn on the tissue and a third degree burn was observed locally in the muscle and skin layers after classic fire needling treatment. This confirms that hwa-acupuncture therapies do not cause major burns. According to the safety assessment test result, no cytotoxicity was detected in the warming fire needling materials. This confirms the safety of the acupuncture materials Conclusions : Various research results on the biological safety of fire needling. Since fire needling therapy induces a burn locally without leaving any scar, and as other results indicate, it is considered a safe treatment method.