• Title/Summary/Keyword: MTS Assay

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A5E promotes Cell growth Arrest and Apoptosis in Non Small Cell Lung Cancer

  • Bak, Ye Sol;Ham, Sun Young;O, Baatartsogt;Jung, Seung Hyun;Choi, Kang Duk;Han, Tae Young;Han, Il Young;Yoon, Do-Young
    • Journal of Applied Biological Chemistry
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    • v.57 no.2
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    • pp.113-122
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    • 2014
  • A5E is complex of several medicinal herb ethanol extracts. The aim of this study is investigating the anticancer effect for non-small cell lung cancer. The antitumor effects of A5E on NCI-H460 were examined by regulation of cell proliferation, apoptosis, cell cycle arrest, mitochondrial membrane potential (${\Delta}{\Psi}_m$), and apoptosis-related protein. Cell proliferation was measured by MTS assay. Apoptosis induced by A5E was confirmed by Annexin V-fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) staining, and cell cycle arrest was measured by PI staining. NF-${\kappa}B$ translocation was detected by immunofluorescence and MMP (${\Delta}{\Psi}_m$) was measured by JC-1 staining. The expression of extrinsic pathway molecules such as FasL and FADD were elevated, and procaspase-8 was processed by A5E. In addition, intrinsic pathway related molecules were altered. The Bcl-2 and Bcl-xl levels decreased, Bax increased, and cytochrome C was released. In addition, the mitochondrial membrane potential collapsed, and caspase-3 and poly-(ADP-ribose) polymerase were processed by A5E. Moreover, A5E affected the cellular survival pathway involving phosphatidylinositol 3-kinase (PI3K)/Akt and NF-${\kappa}B$. PI3K and Akt were downregulated, also NF-${\kappa}B$ expression was decreased, and nuclear translocalization was inhibited by A5E. These results suggested that A5E delays proliferation, inhibit cell cycle progression and induce apoptosis in human lung cancer cell. We conclude that A5E is a potential anticancer agent for human lung carcinoma.

Effects of Liriope muscari Water Extracts on the Cell Death and Inflammatory Cytokine Expression of Poly I:C-treated Lung Carcinoma Cells (맥문동 열수 추출물이 Poly I:C를 처리한 폐암세포주의 사멸 및 염증성 사이토카인 발현에 미치는 영향)

  • Kang, Dayeon;Cho, Namjoon;Renchinkhand, Gereltuya;Lee, Bo-Hee;Kim, Eun-Mi;Nam, Myoung Soo;Kim, Keekwang
    • The Korea Journal of Herbology
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    • v.36 no.1
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    • pp.97-102
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    • 2021
  • Objectives : Virus infection through the respiratory tract causes various inflammatory diseases such as pneumonia, cystic fibrosis, and obstructive pulmonary disease, causing enormous social damage. Therefore, it is very important to develop a treatment and prevention of infectious diseases. In this study, we investigated the effect of water extracts of Liriope muscari (WELM), known to improve lung function, on the inflammatory response of lung carcinoma cell line A549 cells induced by the viral double stranded RNA mimetic Polyinosinic:polycytidylic acid (Poly I:C). Methods : The cell viability by WELM treatment was analyzed using MTS assay in A549 cells. After inducing an inflammatory response to WELM-treated A549 cells with Poly I:C, the degree of apoptosis was confirmed through bright field microscopy. Interferon beta (IFN-β) mRNA expression level in A549 cells was analyzed by quantitative reverse transcription PCR (qRT-PCR). Results : WELM treatment has no significant effect on cell viability of A549 cells. We confirmed that pre-treatment of WELM effectively reduces the Poly I:C-induced apoptotic cell death in A549 cells. In addition, it was confirmed that the mRNA expression level of IFN-β, a pro-inflammatory cytokine increased by Poly I:C treatment, was significantly suppressed by WELM treatment in A549 cells. Conclusions : These results provide the evidence that WELM is effective at inhibiting inflammation on respiratory viral infections and suggest that Liriope muscari might be a valuable natural substance in the prevention and treatment of infectious diseases.

Anti-inflammatory Effect of Bodusan (보두산(寶豆散) 메탄올 추출물의 항염증 효과)

  • Kim, Pan-Joon;Yun, Hyun-Jeong;Heo, Sook-Kyoung;Kim, Kyoung-Ae;Kim, Dong-Wan;Kim, Jae-Eun;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.24 no.2
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    • pp.49-56
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    • 2009
  • Objectives : Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Bodusan (BDS) was a traditional Korean herbal medicine and widely used in treatment of gastrointestinal complaint and stomach ulcer. The aim of this study was to determine whether BDS and its components inhibit production of nitrite, PGE2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods : Cytotoxic activity of BDS and its components on RAW 264.7 cells was using 5-(3caroboxymcrophages. eth-oxyphenyj)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Our results indicated that BDS and its components significantly inhibited the LPS-induced NO and PGE2 production. Moreover. BDS and its components inhibited iNOS and COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-11${\beta}$, IL-6 and MCP-1 formation in macrophages. Conclusions: These results indicate that BDS and its components have potential as an anti-inflammatory agent.

In Vitro Evaluation of Anti-cancer Properties of Hongyoung on SNU-80 Anaplastic Thyroid Carcinoma Cell Line

  • Gaeun Kim;Eun-Jung Kim
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.321-329
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    • 2023
  • Anaplastic thyroid cancer has the highest mortality rate of all thyroid cancers and shows low responsiveness to most treatments. Hongyoung, a reddish-colored potato, is an excellent source of dietary polyphenol containing a large amount of anthocyanins, which has anti-cancer and anti-inflammatory effects. This study investigated the effects of Hongyoung extract on apoptosis and invasiveness in SNU-80 anaplastic thyroid cancer cells. The quantification of the total polyphenol content was done by spectrophotometric measurement. Cell growth was measured by using 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl) 2H tetrazolium, monosodium salt (MTS) assay. Cell cycle was analyzed through FACS analysis. Induction of apoptosis in cells was investigated by annexin V staining using flow cytometer and the expression of caspase-3 and Poly (ADP-ribose) polymerase (PARP) through western blot. mRNA expression and protein activation of matrix metalloproteinases (MMP)-2/-9 were examined by RT-PCR and zymography. As a result, the TPC of Hongyoung was 292.43±8.42 mg gallic acid equivalent (GAE)/100 g dry extract. Hongyoung showed a dose-dependent cell growth inhibition, and the IC50 values was 1,000 ㎍/mL. sub-G1 phase was more than doubled compared to the control group, and S and G2/M phase arrest were also induced. Hongyoung induced apoptosis by increasing FITC-Annexin V-positive cells and increased the activation of caspase-3 (cleaved caspase-3) and PARP (fragmented PARP). Hongyoung significantly inhibited mRNA expression and protein activation of MMP-2/-9 in phorbol 12-myristate 13-acetate (PMA)-treated SNU-80 cells. Therefore, this study suggests the possibility of development of Hongyoung extract as an anti-cancer agent.

Osteoblastogenic Activity of Locusta migratoria Ethanol Extracts on Pre-Osteoblastic MG-63 Cells (풀무치 에탄올 추출물이 MG-63 조골세포 분화에 미치는 영향)

  • Baek, Minhee;Seo, Minchul;Lee, Joon Ha;Kim, In-Woo;Kim, Mi-Ae;Hwang, Jae-Sam
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1448-1454
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    • 2018
  • Insects have been investigated as a novel source of food and biomaterial in several recent studies. However, their osteoblastogenic cell activity has not been sufficiently researched and so, to investigate the potential of this natural material for promoting osteoblastogenesis, we studied the activity of Locusta migratoria ethanol extract (LME) on MG-63 pre-osteoblast cells. The cytotoxicity and proliferation effects of LME on MG-63 cells were measured by MTS assay, and there was no cytotoxicity up to $1,000{\mu}g/ml$. With LME treatment of 500 and $1,000{\mu}g/ml$ for 48 hr, cell proliferation increased to 105% and 116% versus control, respectively. The osteoblastogenic activity of the LME was measured through alkaline phosphatase (ALP) staining at three and five days. As a result, both 500 and $1,000{\mu}g/ml$ LME concentrations were seen to increase ALP activity by more than three times compared with control at three and five days. In addition, the expression level of the osteogenic markers ALP and RUNX2 was markedly increased after LME treatment. These results demonstrate that Locusta migratoria ethanol extract promotes osteoblastogenesis as evidenced by the increased osteogenic markers and suggest that LME may be a potential agent for bone formation and osteoporosis prevention.

Osteoblastogenic Activity of Tenebrio molitor Larvae Oil on the MG-63 Osteoblastic Cell (갈색거저리 유충 오일이 MG-63 조골세포 분화에 미치는 영향)

  • Seo, Minchul;Baek, Minhee;Lee, Joon Ha;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1027-1033
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    • 2019
  • Recently, Korea has seen a rapid increase in the elderly population. As a result, osteoporosis, a geriatric disease, has become a social problem. To investigate the novel and natural materials for promoting osteoblastogenesis, we investigated the osteoblastogenic activity of Tenebrio molitor larvae oil (TMO) on the MG-63 preosteoblast cells. The cytotoxicity and proliferation effects of TMO on MG-63 cells were measured by MTS assay. There was no cytotoxicity up to $80{\mu}g/ml$. At 40 and $80{\mu}g/ml$ of TMO (treated for 48 hr), cell proliferation was elevated about 120% compared to the control. The osteoblastogenic activity of TMO was measured with alkaline phosphatase (ALP) activity at 5 days. Doses of 5 to $80{\mu}g/ml$ of TMO increased ALP activities significantly compared with the control. In addition, expression of ALP and Runx2 (osteoblastogenic markers) were markedly increased after treatment of TMO for 5 days. These results provide evidence that TMO promotes osteoblastogenesis by increasing the gene and protein expression of ALP and Runx2, and they suggest that TMO may be a potential agent for bone formation and preventing osteoporosis.

Whitening and Antioxidant Effects of Extracts from Angelica gigas Nakai Thin Root (참당귀 세미 추출물의 미백 및 항산화 효과)

  • Lee, Sang-Hoon;Park, Jeong-Yong;Seo, Kyung Hye;Choi, Jang Nam;Lee, Yoon-Jeong;Moon, Youn-Ho;Hur, Mok;Park, Woo Tae;Huh, Yun Chan;Chang, Jae-Ki;Koo, Sung Cheol
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.37-43
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    • 2021
  • Angelica gigas Nakai (AGN) is a perennial plant belonging to the family Apiaceae. Its root has been utilized as a traditional medicine especially in Korea. This study was carried out to evaluate the potential use of extracts from AGN root parts as a cosmetic material. The dried AGN roots are divided into body (B), thick root (TkR), medium root(MR) and thin root (TnR) according to their diameter before cutting into medicine. B, TkR and MR of AGN are combined and used as medicinal herbs (MH). The extracts from AGN each root part (B, TkR, MR, TnR, MH) were used to test the effect on cell viability using MTS assay and to examine inhibitory effect on melanin accumulation in B16F10 melanoma cells. All extracts (50 - 200 ㎍/mL) from the each root part did not affect the cell viability. And inhibitory effect of all root extracts (200 ㎍/mL) on melanin accumulation was 12-19%. Especially, TnR showed similar inhibitory effect on melanin accumulation to MH. In addition, DPPH and ABTS free radical scavenging activity were higher in the TnR extract compared to MH. This study showed that the TnR extract exhibit high inhibitory effect on melanin accumulation and antioxidant activity compared to MH, suggesting that TnR extract has potential as a cosmetic ingredient.

Anti-oxidative and Anti-inflammatory Effects of Draconis Resina in Mouse Macrophage Cells (마우스 대식세포에서 혈갈(血竭)(Draconis Resina)의 항산화 및 항염증 효과)

  • Yi, Hyo-Seung;Heo, Sook-Kyoung;Yun, Hyun-Jeong;Choi, Jae-Woo;Jung, Jae-Ha;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.23 no.2
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    • pp.179-192
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    • 2008
  • Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

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Growth Inhibition of Human Uterine Leiomyoma Cells Using Haeohyul-tang (하어혈탕(下瘀血湯)이 1차배양된 인체자궁근종세포(人體子宮筋腫細胞)에 미치는 영향)

  • Kim, Han-Kyun;Zhao, Rong-Jie;Jo, Mi-Jeong;Choi, Sun-Mi;Park, Sook-Jahr;Kim, Mi-Ryeo;Kwon, Young-Kyu;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.158-164
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    • 2007
  • Uterine leiomyoma is the most common tumor in the female genital tract. Although the tumor is benign, it is a matter of paramount importance since it often causes profuse menstrual bleeding, pressure symptoms and infertility. Nevertheless, the etiology and pathophysiology of this abnormality remain poorly understood. The traditional definitive treatment for uterine leiomyomas is hysterectomy and, even today, symptomatic leiomyomas are the leading cause of hysterectomy in Korea. Clearly, the development of a safe, effective, and nonsurgical method of treatment for leiomyoma would be of great benefit to many women. This study demonstrated growth inhibition of uterine leiomyoma cells using Haeohyultang (HT). When human leiomyoma cells were treated with Haeohyultang, cells showed dose-dependent growth inhibitory effect. Cell growth was inhibited by over 40% as determined by both cell counts and MTS assay. Reduction of cellular viability as a consequence of exposure to Haeohyultang resulted from induction of apoptosis, as assessed by DNA fragmentation, PARP cleavage, caspase 9 and caspase 3 assay. Flow cytometry analysis with uterine leiomyoma cells demonstrated sub G1 cell cycle arrest after treatment with drug Haeohyultang. But, the expression levels of p27 and p21 were not changed in Haeohyultang treated cells compared with control. However, the expression levels of clAP1 were reduced by Haeohyultang compared with control. This reduction of clAP1 data means activation of the caspase family, and then induction of PARP cleavage and apoptosis. These results suggest that Haeohyultang may be potential therapeutic approach in the clinical management of uterine leiomyoma.

The Study on Anti-obesity Effects of Gamiygin-tang Extract and Ferment (가미이진탕(加味二陳湯) 전탕액과 발효액이 항비만(抗肥滿)효과에 미치는 연구)

  • Chang, Sung Jin;Min, Deul Le;Park, Eun Jung
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.4
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    • pp.108-121
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    • 2013
  • Objective This study was designed to investigate the effects of Gamiygin-tang (GY) extract (GYE) and fermented solution (GYF) on body weight, serum lipid level and adipocyte differentiation in high fat diet-fed obese mice. Materials and Methods High fat diet-fed obese mice and 3T3-L1 adipocytes mice were treated with GYE and GYF and obesity related markers were assessed. A cytotoxicity assay was carried out by MTS assay. Inhibitory effects of GYE and GYF on adipocyte differentiation were carried out by Oil Red O staining. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors, peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer binding protein alpha (CEBP-${\alpha}$) were measured by real-time reverse transcriptase-polymerase chain reaction. The effects of GYE and GYF on the expression of adipocyte differentiation regulatory factors were also determined in relation to protein production/protein levels by western blotting. The anti obesity effects of GYE and GYF were measured in high fat-diet induced obese mice. Various factors were measured from the serum of the high fat-diet mice. Results Though GYE did not show toxicity at the concentration of 1mg/ml, GYF showed toxicity at the concentration of 1mg/ml. The GYE at 0.1 and 1mg/ml inhibited the differentiation of 3T3-L1 cells, and the GYF also inhibited the differentiation of 3T3-L1 cells. The effect of GYE on adipocyte differentiation factors including PPAR-${\gamma}$ and CEBP-${\alpha}$ was investigated and compared to the corresponding concentration levels of GYF. GYE and GYF both suppressed the RNA and protein levels of adipocyte differentiation factors. In the animal test both GYE and GYF reduced weight gain. GYE and GYF reduced blood cholesterol, TG and LDL levels. GYF better reduced blood cholesterol levels. Conclusions These results demonstrate that GYE and GYF exerts anti-obesity effect in 3T3-L1 cells and obese mice induced by high-fat diet.