• Nutrition Research and Practice
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• 제17권5호
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• pp.899-916
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• 2023

BACKGROUND/OBJECTIVES: Oxidative stress is a fundamental neurodegenerative disease trigger that damages and decimates nerve cells. Neurodegenerative diseases are chronic central nervous system disorders that progress and result from neuronal degradation and loss. Recent studies have extensively focused on neurodegenerative disease treatment and prevention using dietary compounds. Heseperetin is an aglycone hesperidin form with various physiological activities, such as anti-inflammation, antioxidant, and antitumor. However, few studies have considered hesperetin's neuroprotective effects and mechanisms; thus, our study investigated this in hydrogen peroxide (H₂O₂)-treated SH-SY5Y cells. MATERIALS/METHODS: SH-SY5Y cells were treated with H₂O₂ (400 µM) in hesperetin absence or presence (10-40 µM) for 24 h. Three-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays detected cell viability, and 4',6-diamidino-2-phenylindole staining allowed us to observe nuclear morphology changes such as chromatin condensation and apoptotic nuclei. Reactive oxygen species (ROS) detection assays measured intracellular ROS production; Griess reaction assays assessed nitric oxide (NO) production. Western blotting and quantitative polymerase chain reactions quantified corresponding mRNA and proteins. RESULTS: Subsequent experiments utilized various non-toxic hesperetin concentrations, establishing that hesperetin notably decreased intracellular ROS and NO production in H₂O₂-treated SH-SY5Y cells (P < 0.05). Furthermore, hesperetin inhibited H₂O₂-induced inflammation-related gene expression, including interluekin-6, tumor necrosis factor-α, and nuclear factor kappa B (NF-κB) p65 activation. In addition, hesperetin inhibited NF-κB translocation into H₂O₂-treated SH-SY5Y cell nuclei and suppressed mitogen-activated protein kinase protein expression, an essential apoptotic cell death regulator. Various apoptosis hallmarks, including shrinkage and nuclear condensation in H₂O₂-treated cells, were suppressed dose-dependently. Additionally, hesperetin treatment down-regulated Bax/Bcl-2 expression ratios and activated AMP-activated protein kinase-mammalian target of rapamycin autophagy pathways. CONCLUSION: These results substantiate that hesperetin activates autophagy and inhibits apoptosis and inflammation. Hesperetin is a potentially potent dietary agent that reduces neurodegenerative disease onset, progression, and prevention.

• Advances in Traditional Medicine
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• 제1권2호
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• pp.45-51
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• 2000

This study was carried out to evaluate cytotoxicity of the methanol extract from Sophora flavescens Ait. against L1210 (lymphocytic leukemia) and $P388D_1$ (lymphoid neoplasma) Cells in vitro. We have determined cytotoxicity by the MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H- tetrazolium bromide) assay. The order of cytotoxicity of Sophora flavescens Ait. extracts against L1210 and $P388D_1$ cells in vitro is as follows: Fr. 4 > Fr. 3 > Fr. 5 > Fr. 2 > Fr. 1. These results suggest that the fraction 4 of the methanol extracts from Sophora flavescens Ait. may be a valuable choice for the development of antitumor agents. In order to develop an antimicrobial agent, dried Sophora flavescens Ait. was extracted with hot methanol, and then antimicrobial activity (MIC test) was investigated. In this study, the fraction 3 of the methanol extracts from the roots of S. flavescens showed strong growth inhibition activity against gram-positive and gram-negative bacteria (MIC, $3.125\;{\mu}g/ml$) such as S. mutans, S. epidermidis and P. putida. These results indicate that fractions 3 and 4 inhibit tumor cells and bacteria.

• 대한예방한의학회지
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• 제5권2호
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• pp.122-128
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• 2001

This study was carried out to evaluate antitoxic effects of Houttuynia cordata extract on cadmium by colorimetric methods. The antitoxic activity ofc in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. The concentration of 10-2 mg/ml of Houttuynia cordata extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that the ethyl acetate extract of Houttuynia cordata retains a potential antitoxic activity.

• 한국식품영양학회지
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• 제13권5호
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• pp.460-464
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• 2000

본 연구에서는 전통적으로 담석증, 신장치료 등의 한약제로 많이 사용하는 대왕을 여러 용매를 사용하여 얻은 대왕추출물 분액에 대한 세포독성을 여부를 MTT 정량법, NR 정량법, SRB 정량법을 이용하여 조사하였다. 1. 추출 용매 methylene chloride, ethyl acetate, butanol, water로부터 얻은 대왕추출물 모두 처리농도에 따라 세포에 미치는 영향이 증가하였다. 2. Butanol을 용매로 사용하여 얻은 대왕추출물 분액이 다른 3가지 용매로부터 얻은 대왕추출물보다 세포에 미치는 영향이 크게 나타났고 water를 용매로 사용하여 얻은 추출물이 A498 세포주에 미치는 영향이 가장 낮게 나타났다. 3. Butanol을 추출 용매로 하여 얻은 대왕추출물이 A498 세포주에 미치는 영향이 가장 컸는데 그 추출물에 대한 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.63mg/ml, 0.65mg/ml, 0.68mg/ml이었고, 가장 영향이 적은 water의 경우 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.84mg/ml, 0.82mg/ml, 0.80mg/ml이었다. 4. 정량방법 간의 대왕추출물에 대한 반응은 MTT 정량법이 가장 민감하게 나타났다.

• 생약학회지
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• 제31권1호
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• pp.51-56
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• 2000

This study was carried out to evaluate cytotoxic effects of the roots of Sophora flavescens Ait. extracts on murine leukemia tumor cells lines $(P388D_1\;and\;L1210)$. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The comparison of $IC_{50}$ values of the ethyl acetate of Sophora flavescens Ait. extract in leukemia cell lines showed that their susceptibility to these extracts decreased in the following order : Adriamycin>Fr.4>Fr.5>Fr.3>Fr.1>Fr.2 by the MTT assay. These results suggest that the fraction 4 of the ethyl acetate soluble extract of Sophora flavescens Ait. may be a valuable choice for the studies on the treatment of murine leukemia cell lines.

• 한국진공학회지
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• 제22권2호
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• pp.55-65
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• 2013

본 연구에서는, 동시에 넓은 면적을 조사할 수 있는 형태의 유전체 장벽 방전을 이용한 대기압 면방전 플라즈마 발생장치를 이용하여 곰팡이의 살균 실험을 수행하였다. 실험에 사용한 면방전 플라즈마 발생장치의 파워는 사인파 교류전원을 이용하였으며, 1.4~2.3 kV의 방전전압을 가진다. 또한, 유전체 전기용량의 특성으로 인하여 전압과 전류의 위상차는 약 80도를 갖는다. 생물시료에 미치는 온도의 영향은 공랭식 쿨러를 사용하여 유전체의 열을 배출함으로써 최소화 하였으며, 시료의 온도는 온도측정장치를 사용하여 쿨러(Cooler) 작동 시 최대 10분간 플라즈마를 발생시켜도 37도가 넘지 않음을 확인하였다. 플라즈마에서 생성되는 활성종중 오존($O_3$)의 양은 플라즈마 발생부로부터 1 cm 이내에서 약 25~30 ppm이 측정되었으며, 150 cm 떨어진 지점에서도 5 ppm 정도 측정되었다. 그에 비해 일산화질소(NO)나 이산화질소($NO_2$)는 거의 검지되지 않음을 확인하였다. 증류수 속에 담긴 빵 곰팡이 포자를 면방전 플라즈마 발생장치로 처리하였을 때, 포자의 발아율은 처리시간 및 출력파워가 증가함에 따라 급격히 감소하였으며, MTT (3-(4,5-dimethy lthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) 측정법을 통해 분석한 미토콘드리아 활성도도 처리시간과 출력파워에 비례하여 감소함을 보았다. 반면 포자를 Vogel's minimal 배양액에 넣고 플라즈마 처리를 하면, 앞의 실험과 달리 살균효과가 미비함을 보였는데, 이를 통해 포자를 둘러싸고 있는 환경이 플라즈마의 살균효과에 영향을 미치는 것으로 보여진다. 본 연구를 통하여 유전체 장벽을 이용한 면방전 플라즈마 발생장치는 플라즈마 제트(jet)와는 달리 직접적인 접촉 없이도 미생물 살균이 가능하다는 것을 보여주었으며, 이는 면방전 플라즈마 발생장치로부터 발생하는 활성종들이 곰팡이와 같은 미생물의 비활성화에 주요역할을 하기 때문이라고 본다.

• 한방재활의학과학회지
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• 제31권4호
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• pp.1-11
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• 2021

Objectives Hwanggeumjakyak-tang (HJT) has traditionally been used to treat gastrointestinal inflammatory diseases; however, its protective effects against neuronal inflammation are still undiscovered. Methods We investigated the anti-neuroinflammatory effects of HJT water extract on lipopolysaccharide (LPS)-stimulated BV2 mouse microglia cells. BV2 cells were treated with LPS (1 ㎍/mL) 1 hour prior to the addition of HJT. We measured cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and nitrite production using the Griess assay. We performed a reverse transcription-polymerase chain reaction assay to measure messenger RNA expression of inflammatory cytokines including interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. Western blot analysis was performed to determine protein expression of mitogen-activated protein kinases (MAPKs) and inhibitor of nuclear factor kappa B (NF-κB)α. Results HJT inhibited excessive nitrite release in LPS-stimulated BV2 cells and also significantly inhibited inflammatory cytokines such as IL-1β, IL-6, and TNF-α in LPS-stimulated BV2 cells. Moreover, HJT significantly suppressed LPS-induced MAPK and NF-κB activation and inhibited the elevation of IL-1β, IL-6, and TNF-α in the brain of LPS-injected mice. Conclusions Our study highlights the anti-neuroinflammatory effects of HJT via MAPK and NF-κB deactivation.

• Restorative Dentistry and Endodontics
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• 제48권1호
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• pp.2.1-2.12
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• 2023

Objectives: In this study, natural substances were introduced as primary dental pulp caps for use in pulp therapy, and the antimicrobial and cytotoxic properties of these substances were investigated. Materials and Methods: In this in vitro study, the antimicrobial properties of calcium-enriched mixture (CEM) cement, propolis, and propolis individually combined with the extracts of several medicinal plants were investigated against Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Then, the cytotoxicity of each substance or mixture against pulp stem cells extracted from 30 primary healthy teeth was evaluated at 4 concentrations. Data were gathered via observation, and optical density values were obtained using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) test and recorded. SPSS software version 23 was used to analyze the data. Data were evaluated using 2-way analysis of variance and the Tukey test. Results: Regarding antimicrobial properties, thyme alone and thyme + propolis had the lowest minimum inhibitory concentrations (MICs) against the growth of S. aureus, E. coli, and P. aeruginosa bacteria. For E. faecalis, thyme + propolis had the lowest MIC, followed by thyme alone. At 24 and 72 hours, thyme + propolis, CEM cement, and propolis had the greatest bioviability in the primary dental pulp stem cells, and lavender + propolis had the lowest bioviability. Conclusions: Of the studied materials, thyme + propolis showed the best results in the measures of practical performance as a dental pulp cap.

• Journal of Veterinary Science
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• 제25권2호
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• pp.21.1-21.15
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• 2024

Background: Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial. Objectives: To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication. Methods: The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway. Results: The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly downregulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010. Conclusions: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.

• 접착 및 계면
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• 제10권2호
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• pp.77-83
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• 2009

생체적용 센서 코팅 재료로서 polydimethylsiloxane (PDMS)를 선정하였으며 합성 및 성형과정에서 용출될 수 있는 잔류 독성 물질의 세포독성을 확인하고자 하였다. ISO 10993-5, Biological evaluation of medical devices-Part 5 : Tests for in vitro cytotoxicity (의료기기의 생물 안정성 평가-제5부: 세포 독성 시험-체외시험)를 통하여 세포 독성 평가를 실시하였다. 양성 대조군으로 organo-tin을 사용하였으며 음성 대조군으로 혈청이 포함되지 않은 RPMI 1640배지를 사용하였다. 고체 시료의 표면적에 대하여 $125{\mu}L/cm^2$가 되도록 혈청이 포함되지 않은 RPMI 1640 배지를 용기에 첨가하였으며 $38^{\circ}C$를 유지하며 일정시간 동안 추출하였다. 세포 독성 평가는 1) NIH 3T3 fibroblast 단일세포층을 형성한 후 추출물을 첨가하는 방법과 2) 세포와 함께 추출물을 넣어 배양하는 방법을 동시에 시행하였다. 세포 형태학적인 변화 관찰과 MTT (tetrazolium dye, 2-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) 시험법에 의한 세포 활성 측정을 병행함으로써 고체 시료로부터 추출된 물질의 세포독성 여부와 고체시료의 표면에 대한 세포의 감응성도 함께 관찰할 수 있었다.