• 제목/요약/키워드: MTT assay

검색결과 2,655건 처리시간 0.028초

PAMAM Dendrimers Conjugated with L-Arginine and γ-Aminobutyric Acid as Novel Polymeric Gene Delivery Carriers

  • Son, Sang Jae;Yu, Gwang Sig;Choe, Yun Hui;Kim, Youn-Joong;Lee, Eunji;Park, Jong-Sang;Choi, Joon Sig
    • Bulletin of the Korean Chemical Society
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    • 제34권2호
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    • pp.579-584
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    • 2013
  • In this study, we synthesized functional dendrimer derivatives as nonviral gene delivery vectors. Poly(amidoamine) dendrimer (PAMAM, generation 4) was modified to possess functional amino acids to enhance gene transfection efficiency. PAMAM G4 derivatives conjugated with L-arginine (Arg) and ${\gamma}$-aminobutyric acid (GABA) showed higher transfection efficiency and lower cytotoxicity compared to the native PAMAM G4 dendrimer. The polyplex of the PAMAM G4 derivative/pDNA was evaluated using an agarose gel retardation assay and Picogreen reagent assay. Additionally, the MTT assay was performed to examine the cytotoxicity of synthesized polymers. All PAMAM G4 derivatives showed lower cytotoxicity than PEI25kD. Particularly, PAMAM G4-GABA-Arg displayed enhanced transfection efficiency compared to the native PAMAM G4 dendrimer.

유근피 추출물 및 분획의 생리활성 (Physiological Activity of Extracts of Bark from Ulmus davidiana var. japonica and its fractions)

  • 양선아;김아영;표병식;김선민
    • 생약학회지
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    • 제50권2호
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    • pp.112-117
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    • 2019
  • In this study, extracts of bark from Ulmus davidiana var. japonica and its fractions were investigated the antioxidative, antibacterial and tyrosinase inhibition activity for physiological activity towards functional applications. In the measurement of DPPH radical scavenging activity, the ethyl acetate fraction showed higher radical scavenging ability than others. Moreover, in the tyrosinase inhibition assay, showed that the ethyl acetate fraction has good inhibition effects. Results of the DPPH radical scavenging and tyrosinase inhibition activity are related with the total polyphenol concentrations of ethyl acetate fraction. In antibacterial activity used to find out by utilizing the disc diffusion assay, chloroform fraction showed strong effect against Staphylococcus aureus and S. epidermidis. These results are related with the flavonoid contents of chloroform fraction. On the other hand, in the L929 cell viability measurement by MTT assay, the hexane, butanol and aqueous fraction treated at high concentration were showed cytotoxicity. But the others samples were exhibited a moderate viabilities. As a result of investigated the antioxidant and tyrosinase inhibition activity, the ethyl acetate fraction could be applicable for cosmetics related fields. And the chloroform fraction showed significant antibacterial activity against S. aureus and S. epidermidis.

Inhibitory Effects of Ginsenosides on Glutamate-Induced Swelling of Cultured Astrocytes

  • Seong, Yeon-Hee;Koh, Sang-Bum;Kim, Hack-Seang
    • Journal of Ginseng Research
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    • 제24권3호
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    • pp.138-142
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    • 2000
  • 흰쥐 대뇌겉질로부터 별아교세포를 배양하여 흥분성아미노산인 L-glutamate에 의하여 유발되는 세포종창(astrocytic swelling)에 대한 ginsenosides의 억제효과를 검토하였다. Glutamate(0.5 mM)를 세포에 가하고 1시간동안 배양하면 swelling을 일으켜, 세포내의 물의 용적([$^3$H]OMG의 uptake량으로 측정)은 대조세포에 비하여 약 2배의 증가를 나타냈다. Glutamate와 함께 ginsenosides Rb$_2$와 Rc를 가하고 배양하면 glutamate에 의한 astrocytic swelling이 용량의존적으로 감소하였다. 세포는 Rb$_2$와 Rc(0.5 mg/ml)에 24시간까지 노출시켜도 MTT reduction이 감소하지 않는 것으로 보아 이 ginsenosides에 의한 swelling의 억제효과는 세포막의 손상에 의한 것이 아님을 알 수 있었다. Rb$_2$와 Rc는 glutamate에 의한 세포내 $Ca^{2+}$농도의 상승을 억제하였다. 따라서 Rb$_2$와 Rc는Ca$^{2+}$의 유입을 억제하므로서 glutamate에 의한 astrocytic swelling을 억제하는 것으로 생각된다.

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Paclitaxel에 의한 관절연골 세포의 capase-비의존적 mitotic catastrophe 유도 (Paclitaxel Induced Caspase-Independent Mitotic Catastrophe in Rabbit Articular Chondrocyte)

  • 임정희;김송자
    • 생명과학회지
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    • 제20권4호
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    • pp.519-527
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    • 2010
  • Paclitaxel은 미세소관의 탈중합을 억제하는 시약으로 알려져 있다. Paclitaxel은 다양한 세포에서 세포 내 방추체를 안정화시킴으로써 유사분열 억제 및 세포사멸을 유도한다. 본 실험에서는 토끼 관절 연골세포에서 paclitaxel이 연골세포의 증식과 사멸에 미치는 효과에 대한 연구를 수행하였다. MTT assay를 수행한 결과 paclitaxel은 연골세포에서 농도 의존적으로 세포 증식을 억제한다는 것을 확인 할 수 있었으며, FACS analysis와 Western blot analysis를 수행한 결과, paclitaxel이 G2/M 정지를 유도하는 것을 확인하였다. 또한, paclitaxel이 비정상적인 세포 분열유도와 핵 단편분절 유도없이 일어나는 mitotic catastrophe 즉, caspase-3 비의존적인 세포사멸을 유도하였다. Paclitaxel을 처리한 세포에서 일어나는 이러한 mitotic catastrophe에 의한 세포 죽음은 G1/S기의 진행을 억제하는 시약인 thymidine을 처리하는 것에 의해 억제되는 것을 확인할 수 있었다. 이러한 결과를 종합해 볼 때, paclitaxel에 의한 토끼 관절 연골 세포에서의 세포 죽음은 caspase-3 비의존적인 mitotic catastrophe에 의해 일어나는 것으로 사료되어진다.

가미보중익기탕이 GLUCOSE OXIDASE에 의해 손상된 배양 척수감각신경세포의 총단백질 합성량에 미치는 영향 (Effects of Gamibojungikki-tang on Total Protein Synthesis of Cultured Spinal Sensory Neurons Damaged by GLUCOSE OXIDASE)

  • 이창호;권강범;장승호;송용선;류도곤
    • 동의생리병리학회지
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    • 제16권1호
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    • pp.141-145
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    • 2002
  • In order to clarify the neuroprotective effect of Gamibojungikki-tang (GBJIKT) water extract on cultured mouse spinal sensory neuron damaged by glucose Oxidase (GO), MTT [3-(4,5-dimethylthiazole-2-yl) -2,5-diphenyltetrazolium bromide] assay and SRB (Sulforhodamine B) assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of GBJIKT water extract for 3 hours prior to exposure of GO. Cell viability of cultured mouse spinal sensory neurons exposed to various concentrations of GO for 8 hours was decreased in a dose-dependent manner. MTT50 values were 45 mU/ml GO. Cultured mouse spinal sensory neurons in the medium containing various concentration of GO for 8 hours showed decreasing of total protein synthesis. GO was toxic on cultured spinal sensory neurons. Pretreatment at GBJIKT water extract for 3 hours following GO prevented the GO-induced neurotoxicity such as decreasing of total protein synthesis. These results suggest that GO shows toxic effect on cultured spinal sensory neurons and GBJIKT water extract is highly effective in proecting the neurotoxicity induced by GO.

조구등이 Glucose Oxidase로 손상된 대뇌신경세포에 미치는 효과 (Effect of Ramulus et uncus uncariae on Glucose Oxidase-Induced Toxicity in Cultured Cerebral Neurons)

  • 김형수;이용석;오석규;이강창;이건목;이정원;이상복;김종호;유준기;강영성;김성수;송호준;박승택
    • 동의생리병리학회지
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    • 제16권5호
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    • pp.1016-1019
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    • 2002
  • To examine the cytotoxic effect of glucose oxidase(GO) in cultured mouse cerebral neurons, cytotoxicity was measured by MTT assay after cultured nerve cells were incubated for 3 hours in the media containing 1 ~ 60mU/ml concentrations of GO. In addition, the neuroprotective effect of Ramulus et uncus uncariae(REUU) was determined by MTT assay in these cultrures. Cell viability was remarkably decreased in a dose- and time-dependent manner after cultured mouse cerebral neurons were exposed to 30mU/ml GO for 3 hours. In the neuroprotective effect of REUU on GO-induced toxicity, REUU blocked the GO-mediated neurotoxicity in these cultures. From above the results, it suggests that GO is toxic in cultured mouse cerebral neurons and selective herb extract such as REUU is effective in prevetion of the neurotoxicity induced by GO.

비알코올성 지방간 세포 모델에 대한 택사, 산사, 구기자, 울금, 단삼, 인진의 효능 비교 (Comparison of the Therapeutic Efficacy of Rhizoma Alismatis, Fructus Crataegi, Fructus Lycii, Radix Curcumae, Radix Salviae Miltiorrhizae, Herba Artemisiae Scopariae on the Experimental Cellular Model of Nonalcoholic Fatty Liver Disease)

  • 한창우;주명수;이장훈
    • 대한한방내과학회지
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    • 제33권4호
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    • pp.533-542
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    • 2012
  • Objectives : We try to compared the efficacy of six herbal medicines, Rhizoma Alismatis (RA), Fructus Crataegi (FC), Fructus Lycii (FL), Radix Curcumae (RC), Radix Salviae Miltiorrhizae (RSM), and Herba Artemisiae Scopariae (HAS), constituting KHchunggan-tang which was previously proven to be hepatoprotective on non-alcoholic fatty liver disease with combined properties of cellular steatosis, ROS production, and cytoprotection. Methods : HepG2 cells were pretreated with aqueous extracts of the six herb medicines at concentrations of 1, 10, 50 and 100 ${\mu}g/ml$ each, and treated with 0.5 mM palmitate consecutively. After 21 hrs, cell viability was assessed using MTT assay, and the percentage of cells with sub-G1 DNA content was measured using fluorescence-activated cell sorting after propidium iodide staining. Results : The first three extracts, RA, FC, and FL restored cell viability reduced by palmitate in MTT assay, and RA, FC, FL and RC inhibited palmitate-induced apoptosis in sub-G1 analysis. FL showed relatively weak potential only at tested maximal dose, and RA showed the greatest higher efficacy on this experimental cellular model of nonalcoholic fatty liver disease. Conclusions : According to this comparative experiment, Rhizoma Alismatis seems to have the most powerful potential among the six herbs constituting KHchunggan-tang, and consecutive further study seems to be required for more standardized and effective clinical application of KHchunggan-tang for treatment of non-alcoholic fatty liver disease.

Cisplatin Combined with Metformin Inhibits Migration and Invasion of Human Nasopharyngeal Carcinoma Cells by Regulating E-cadherin and MMP-9

  • Sun, Xiao-Jin;Zhang, Pei;Li, Hai-Hui;Jiang, Zhi-Wen;Jiang, Chen-Chen;Liu, Hao
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권9호
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    • pp.4019-4023
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    • 2014
  • Metformin has been shown to be useful in reducing insulin resistance by restoring sensitivity. Recent evidence suggests that metformin might also possess anti-tumour activity. This study aimed to investigate the effects of cisplatin combined with metformin on the proliferation, invasion and migration of HNE1/DDP human nasopharyngeal carcinoma (NPC) cells, and to provide a new target for treating metastasis. The MTT assay was used to assess viability of HNE1/DDP cells after exposure to different concentrations of 2, 5-diaminopyrimidine-4, 6-diol (DDP; 2, 4, 8, 16, and $32{\mu}mol{\cdot}L^{-1}$), metformin (5, 10, 15, 20, and $25{\mu}mol{\cdot}L^{-1}$), and $4{\mu}mol{\cdot}L^{-1}$ of DDP combined with metformin. Wound healing and transwell migration assays were performed to assess cell migration and invasion, and expression of E-cadherin and MMP-9 was detected using Western blotting. MTT assay results showed that DDP could inhibit the proliferation of HNE1/DDP cells in a time- and concentration-dependent manner, with an IC50 of $32.0{\mu}mol{\cdot}L^{-1}$ at 24 h (P < 0.05), whereas low concentrations of DDP had almost no inhibitory effects on cell invasion and migration. DDP combined with metformin significantly inhibited cell invasion and migration. In addition, genes related to migration and invasion, such as those of E-cadherin and MMP-9, showed differential expression in the NPC cell line HNE1/DDP. In the present study, with an increasing concentration of metformin, the expression of MMP-9 was downregulated whereas that of E-cadherin was significantly upregulated. Taken together, our results show that cisplatin combined with metformin has effects on proliferation, invasion, and migration of human NPC cells.

의안의 제조 방법에 관한 연구 (A study of the manual procedure of ocular prosthesis)

  • 김재민;박동화;유근창;김순애;조승권
    • 한국안광학회지
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    • 제7권2호
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    • pp.95-99
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    • 2002
  • 단안을 잃은 사함의 외모를 인위적으로 보상하기 위한 방법으로 의안을 작용하게 되는데 본 연구에서는 의안의 제조 과정 중 가장 기본적인 방법을 소개하고 의안 용출액의 세포독성을 평가하여 의안 제조의 기술 발달을 촉진하고 안경사의 의안에 대한 관심과 연구가 활발해지기를 기대하며 본 연구를 수행하였다. 의안의 제작 과정은 왁스(wax)나 컨포머(conformer)를 이용한 기본형 제작, 석고 본뜨기, 공막 제작, 홍채 및 동공 제작, 각막 제작, 검열반 제작 그리고 연마과정을 거치는데 각 과정에 대하여 필요한 재료와 제작과정을 살펴보았다. 또한 의안의 세포 독성을 검정하기 위해 의안을 용출시켜 얻은 용액을 세포에 직접 처리하여 세포증식 저해정도를 MTT assay로 조사하였다. 본 연구에 이용된 의안의 용출액은 세포독성이 없는 것으로 나타났으며 제작과정은 가장 기본적인 제작과정을 보여줌으로써 앞으로의 의안 연구에 많은 도움을 줄 것으로 사료된다.

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점안액 보존제 성분 Benzalkonium Chloride에 의해 유도된 각막상피세포의 세포고사 유도 (Effects of Benzalkonium Chloride on the growth and survival of Human corneal epithelial cells)

  • 김재민;이석주;서은선
    • 한국안광학회지
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    • 제7권2호
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    • pp.189-195
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    • 2002
  • 점안액 보존제 성분으로 이용되는 Benzalkonium Chloride(BAC)가 배양 각막 상피세포에 미치는 영향을 조사하기 위해 본 연구를 시행하였다. BAC를 세포에 0.0001%~0.01%로 15분 동안 처리하여 24시간 후 회복 효과를 조사하였다. MTT assay를 이용하여 세포 생존율을 산정하고 Hoechst 33342로 염색질 응축을 조사하였다. Fas 발현 여부를 조사하기 위해 western blot과 immunocytochemistry를 이용하였으며 DNA fragmentation은 agarose gel을 이용한 전기영동을 시행하였다. BAC를 0.005% 이상 처리하면 세포괴사가 일어나는 반면에 저농도의 BAC는 세포고사를 유도하는 것으로 나타났으며 BAC로 Fas 발현이 유도될 수 있다는 것을 보여주고 있다. 본 연구를 통해 점안액의 대표적인 보존제인 BAC의 부작용에 대한 각막세포 차원의 연구가 필요하며 세포독성이 없는 새로운 보존제의 개발이 필요한 것으로 나타났다.

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