• Title/Summary/Keyword: MTT assay

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Propranolol Inhibits the Proliferation of Human Glioblastoma Cell Lines through Notch1 and Hes1 Signaling System

  • Kim, Hyun Sik;Park, Young Han;Lee, Heui Seung;Kwon, Mi Jung;Song, Joon Ho;Chang, In Bok
    • Journal of Korean Neurosurgical Society
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    • v.64 no.5
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    • pp.716-725
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    • 2021
  • Objective : The anti-tumor effect of the beta-adrenergic receptor antagonist propranolol in breast cancer is well known; however, its activity in glioblastoma is not well-evaluated. The Notch-Hes pathway is known to regulate cell differentiation, proliferation, and apoptosis. We investigated the effect of propranolol to human glioblastoma cell lines, and the role of Notch and Hes signaling in this process. Methods : We performed immunohistochemical staining on 31 surgically resected primary human glioblastoma tissues. We also used glioblastoma cell lines of U87-MG, LN229, and neuroblastoma cell line of SH-SY5Y in this study. The effect of propranolol and isoproterenol on cell proliferation was evaluated using the MTT assay (absorbance 570 nm). The impact of propranolol on gene expression (Notch and Hes) was evaluated using real-time polymerase chain reaction (RT-PCR, whereas protein levels of Notch1 and Hes1 were measured using Western blotting (WB), simultaneously. Small interfering RNA (siRNA) was used to suppress the Notch gene to investigate its role in the proliferation of glioblastoma. Results : Propranolol and isoproterenol caused a dose-dependent decrease in cell proliferation (MTT assay). RT-PCR showed an increase in Notch1 and Hes1 expression by propranolol, whereas WB demonstrated increase in Notch1 protein, but a decrease in Hes1 by propranolol. The proliferation of U87-MG and LN229 was not significantly suppressed after transfection with Notch siRNA. Conclusion : These results demonstrated that propranolol suppressed the proliferation of glioblastoma cell lines and neuroblastoma cell line, and Hes1 was more closely involved than Notch1 was in glioblastoma proliferation.

Antinociceptive, anti-inflammatory, and cytotoxic properties of Origanum vulgare essential oil, rich with β-caryophyllene and β-caryophyllene oxide

  • Moghrovyan, Armenuhi;Parseghyan, Lilya;Sevoyan, Gohar;Darbinyan, Anna;Sahakyan, Naira;Gaboyan, Monica;Karabekian, Zaruhi;Voskanyan, Armen
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.140-151
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    • 2022
  • Background: Essential oils are of great interest for their analgesic and anti-inflammatory properties. We aimed to study the content of the essential oil of the Origanum vulgare of the Armenian highlands (OVA) in different periods of vegetation and to investigate its antinociceptive and anti-inflammatory effects in mice (in vivo) and cytotoxic action in cultured cells (in vitro). OVA essential oil was extracted from fresh plant material by hydro-distillation. Methods: For OVA essential oil contents determination the gas chromatography-mass spectrometry method was used. Formalin and hot plate tests and analysis of cell viability using the methyl-thiazolyl-tetrazolium (MTT) assay were used. Results: The maximal content of β-caryophyllene and β-caryophyllene oxide in OVA essential oil was revealed in the period of blossoming (8.18% and 13.36%, correspondently). In the formalin test, 4% OVA essential oil solution (3.5 mg/mouse) exerts significant antinociceptive and anti-inflammatory effects (P = 0.003). MTT assay shows approximately 60% cytotoxicity in HeLa and Vero cells for 2.0 µL/mL OVA essential oil in media. Conclusions: The wild oregano herb of Armenian highlands, harvested in the blossoming period, may be considered as a valuable source for developing pain-relieving preparations.

Evaluation of Antiaging Activity on Lycium chinense Miller. Extracts (구기자 추출물의 항노화 활성 평가)

  • Lee, Yong-Hwa;Kim, ․Na-Yeon;Jo, Il-young;Yi, Yong-Sub
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.1
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    • pp.27-33
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    • 2022
  • The purpose of this study was to investigate the functionality of Lycium chinense Miller. as a functional cosmetic material. The total polyphenol content in the extract was 17.3 mg/mL, and as a result of confirming cytotoxicity through MTT assay before the enzyme experiment, it was confirmed that it did not affect cell growth at all concentrations below 1000 ㎍/mL. As a result of conducting DPPH radical scavenging tests as an antioxidant efficacy test, it showed 84.97% scavenging ability at a concentration of 1000 ㎍/mL, and showed very excellent antioxidant efficacy in SOD-like activity test with a result of 80.54% at 1000 ㎍/mL. The measured elastase inhibitory activity, the inhibition rate of 50.93% was observed at 1000 ㎍/mL, indicating that the extract of Lycium chinense Miller. was effective in inhibiting elastase. All these findings suggested that 70% ethanol extract of Lycium chinense Miller. was confirmed as a functional cosmetic material by confirming its antioxidant and anti-wrinkle efficacy.

Physical Properties and Cytotoxicity of Dental Pit and Fissure Sealants Containing Cerium Oxide Nano Particles(CNPs) (세륨옥사이드나노입자(Cerium Oxide Nano Particles; CNPs)를 첨가한 치면열구전색재의 물리적 특성 및 세포독성)

  • Jeong, Mi-Ae;Kim, Dong-Ae
    • The Journal of the Korea Contents Association
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    • v.22 no.3
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    • pp.586-592
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    • 2022
  • In this study cerium nano particles(CNPs) with 0-4.0 wt% was incorporated to the conventional dental pit and fissure sealant(ConciseTM) to produce new pit and fissure sealant the physical properties and cytotoxicity. The physical properties were measured for polymerizing depth the degree of water absorption and solubility. The cytotoxicity of cell viability was analyzed by MTT assay using immortalized human oral keratinocyte(IHOK). As a result of this preceding study the polymerizing depth was decreased by the increasing of the amount of CNPs. The solubility degree of the sealant added CNPs with 2.0 wt% showed was the lower and the water absorption showed no significantly difference with the control groups(p>0.05). The cytotoxicity test results showed high survival rates in all experimental groups. Therefore, pit and fissure sealant by the addition of CNPs excellent cell viability be produced without weaken the physical property of the cell viability fissure sealant containing CNPs does not weaken physical properties and has no cytotoxic effects biocompatibility. Considering its properties effect of CNPs, further studies are required for distribution technology application.

Effects of Liriopis Tuber on 4-HNE-induced Apoptosis in PC-12 Cells (맥문동(麥門冬)이 4-HNE에 의해 유도된 Apoptosis에 미치는 영향)

  • Park, Soo-Hwan;Kim, Youn-Sub
    • The Korea Journal of Herbology
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    • v.28 no.2
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    • pp.33-38
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    • 2013
  • Objects : Apoptosis leads to the death of a cell. The mitochondrial pathway of apoptosis, a process regulated by the Bcl-2 family of proteins, plays a key role in various biological processes. The tuber of Liriope platyphylla Wang et TANG (Liliaceae), also known as Liriopis tuber, is famous in Oriental medicine owing to its tonic, antitussive, expectorant and anti-asthmatic properties. The purpose of this study was to observe the effect of the Liriopis tuber on mitochondrial-mediated apoptosis in PC-12 cells. Method : A cytotoxic test on Liriopis Tuber water extract was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE 25 ${\mu}M$ that causes oxidative stress in PC-12 cells for 24 hours. In addition, in order to observe the expression of Bcl-2 and Bax protein involved with apoptosis, western blot was conducted. Results : The LT water extract had no toxicity for PC-12 cell. In the cytoprotective effect against 4-HNE, both of the group treated with 50 ${\mu}g/m{\ell}$ and 100 ${\mu}g/m{\ell}$ of LT water extract showed a significant increase in comparison with the control group. In Bax protein expression, all the experimental groups treated with LT water extract showed a decrease in comparison with the control group but had no significance. In Bcl-2 protein expression, all the experimental groups treated with LT water extract showed a significant increase in comparison with the control group. Conclusion : These results suggest that LT is effective in reducing apoptosis.

Houttuynia cordata Thunberg exhibits anti-tumorigenic activity in human gastric cancer cells

  • Hong, Se Chul;Eo, Hyun-Ji;Song, Hoon-Min;Woo, So-Hee;Kim, Mi-Kyeong;Lee, Jin-Wook;Seo, Jeong-Min;Park, Su-Bin;Eom, Jung-Hye;Koo, JinSuk;Jeong, JinBoo
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.155-160
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    • 2013
  • Objectives : Gastric cancer is a leading cause of cancer-related deaths, worldwide. Houttuynia cordata Thunberg (H. cordata) has been used as a medicinal plants and it has an anti-cancer activity in human colorectal cancer and leukemic cancer. However, the potential anti-cancer activity and mechanisms of H. cordata for human gastric cancer cells have not been tested so far. Thus, this study examined the biological effects of H. cordata on the human gastric cancer cell line SNU-1 and AGS. Methods : Inhibition of cell proliferation and cell cycle by H. cordata was carried out by MTT assay and Muse cell cycle analysis and the expressions of protein associated with apoptosis and cell cycle regulation were investigated with Western blot analysis. Results : In MTT assay, the proliferation of SNU-1 and AGS cells was significantly inhibited by H. cordata in a time and dose dependent manner, Inhibition of cell proliferation by H. cordata was in part associated with apoptotic cell death, as shown by changes in the expression ratio of Bax to Bcl-2 by H. cordata. Also, H. cordata regulated the expression of cell cycle regulatory proteins such as pRb, cyclin D1, cyclin E, CDK4, CDK2, p21 and p15. Conclusion : The antiproliferative effect of H. cordata on SNU-1 and AGS gastric cancer cells revealed in this study suggests that H. cordata has intriguing potential as a chemopreventive or chemotherapeutic agent.

Anti-inflammatory Effect of the Hedgehog Skin Extracts on LPS-Stimulated RAW 264.7 Cells

  • Kim, Tae Yeon;Jo, Na Young;Leem, Kang Hyun
    • The Korea Journal of Herbology
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    • v.33 no.3
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    • pp.19-24
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    • 2018
  • Objectives : Hedgehog skin is one of the animal medicines in Traditional Korean Medicine for hematochezia and hemorrhoids. In this study, we examined cytotoxicity and anti-inflammatory effects. Methods : Cytotoxicity of hedgehog skin extracts was measured by MTT assay in vitro. We investigated the inhibition of lipopolysaccharide (LPS) stimulated nitric oxide (NO) production in RAW 264.7 cells. The phosphorylation of mitogen-activated protein kinases (MAPKs) was measured by western blot. And we observed the effect of hedgehog skin extracts on the expression of IL-6 genes using real time PCR. Results : As a result of MTT assay for cytotoxicity, there were no significant differences between non-treatment group and hedgehog skin extracts treatment groups. $500{\mu}g/m{\ell}$ of hedgehog skin extracts treatment significantly decreased nitric oxide production in comparison with non-treatment in LPS-induced RAW 264.7 cells. In measurement of the phosphorylation of MAPKs using western blot analysis, LPS stimulation increased the phosphorylation of MAPKs and $500{\mu}g/m{\ell}$ of hedgehog skin extracts treatment decreased the phosphorylation of ERK1, ERK2 and p38 significantly. But there were no significant differences the phosphorylation of JNK1 and JNK2. As a result of confirmation of the IL-6 mRNA gene expression using real time PCR, IL-6 mRNA gene expressions were significantly decreased in $50{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$ hedgehog skin extracts treated groups by comparison with non-treatment group. Conclusion : These results could provide a mechanistic explanation for the anti-inflammatory effects of the hedgehog skin.

Dual Cytotoxic Responses Induced by Treatment of A549 Human Lung Cancer Cells with Sweet Bee Venom in a Dose-Dependent Manner

  • Yu-Na Hwang;In-Seo Kwon;Han-Heom Na;Jin-Sung Park;Keun-Cheol Kim
    • Journal of Pharmacopuncture
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    • v.25 no.4
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    • pp.390-395
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    • 2022
  • Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells. Methods: We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis. Results: Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax. Conclusion: sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

Basic research for exploring anti-obesity activity of several medicinal plants from Mongolia (몽골산 약용식물 수종의 항비만 활성 탐색을 위한 기초 연구)

  • Moon-Yeol Choi;So-Young Kim;Mi Ryeo Kim
    • The Korea Journal of Herbology
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    • v.38 no.3
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    • pp.1-10
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    • 2023
  • Objective : Obesity in modern society has a significant impact on pathological, psychological and social problems. Therefore, many studies on obesity treatment using herbal extracts with fewer side effects have been conducted. This study was designed to investigate the effect of inhibiting fat accumulation in vitro in order to Mongolian medicinal plants find anti-obesity candidate among. Methods : We measured pancreatic lipase inhibitory activity in Mongolian medicinal plants extract. Cytotoxicity of these extracts was monitored in 3T3-L1 cells by MTT assay. In addition, the anti-obesity effects was confirmed at concentrations of 0.2, 0.1 and 0.05 mg/㎖ through Oil red O staining. Results : Among Mongolian medicinal plants, Rheum undulatum roots in September (RURS), Paeonia anomala L. (PAL), and Fragaria orientalis (FO) showed the highest pancreatic lipase inhibitory activity. As a result of the MTT assay, more than 80% was judged to be non-toxic, and the concentration was determined, and as a result of evaluating the lipid accumulation inhibitory effect, 6 types were selected as candidates. Conclusion : Based on these results, the top 7 species expected to be used as anti-obesity functional materials were selected. However, additional efficacy verification and mechanism of action need to be established in the future. So, it is expected that the medicinal plants verified through this will be used as functional materials for the prevention and treatment of obesity.

Antioxidant activity and MMP-1 inhibitory activity of Panax Ginseng Sprout Extracts (새싹인삼 추출물의 항산화 활성 및 MMP-1 저해 활성 )

  • Min-Jung Kim;Ye-Jin Yang;Ju-Hye Yang;Won-Yung Lee;Woo-Hyun Kim;Jae-Nam Lee;Kwang-Il Park
    • Herbal Formula Science
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    • v.32 no.1
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    • pp.83-90
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    • 2024
  • Objectives : As a substitute for high-price ginseng, this study attempted to examine a possibility of the ferment extract of Panax ginseng sprout whether leaves and roots can be used together as a cosmetic ingredient with anti-oxidative and wrinkle-care effects. Methods : In terms of a test method, antioxidant activities were confirmed through total polyphenol contents, total flavonoid contents, DPPH radical scavenging activity and ABTS radical scavenging activity using the Panax ginseng sprout. In addition, to assess wrinkle-care effectiveness, the cytotoxicity of the extract was analyzed through MTT assay, and inhibition of collagenase activity in the cells was tested using the Panax ginseng sprout fermented by Saccharomyces cerevisiae. Resuits : The content of polyphenols and flavonoids in natural plants was highest in Panax Ginseng Sprout Extract at 100℃, which also demonstrated high DPPH, ABTS radical scavenging activity. MTT assay demonstrated that the Panax Ginseng Sprout Ferment Extract did not have a cytotoxic effect in CCD-986SK cell. Also, Panax Ginseng Sprout Ferment Extract was found to inhibit MMP-1 expression by 51.85±6.09% at a concentration of 10%. Conclusions : Therefore, this study has confirmed a possibility of Panax ginseng sprout ferment extract as a cosmetic ingredient with MMP-1-inhibitory effects.