• Title/Summary/Keyword: MTT assay

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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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    • v.34 no.2
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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.

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

  • Yang, Sun a;Kim, A young;Pyo, Byoung sik;Kim, Sun min
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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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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    • v.24 no.3
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    • pp.138-142
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    • 2000
  • Effects of ginsenosides (Rb$_1$, Rb$_2$, Rc, Re, Rg$_1$, Rf) on L-glutamate (glutamate)-induced swelling of cultured astrocytes from rat brain cerebral cortex were studied. Following the exposure to 0.5mM glutamate for 1 hr, the intracellular water space (as measured by [$^3$H]O-methyl-D-glucose uptake) of astrocytes increased by about two-fold. Simultaneous addition of ginsenosides Rb$_2$ and Rc with glutamate reduced the astrocytic swelling in a dose-dependent manner. These ginsenosides at 0.5 mg/ml did not affect the viability of astrocytes for up to 24 hr which was determined by a colorimetric assay (MTT assay) for cellular growth and survival. These ginsenosides at 0.3 mg/ml inhibited the increase of intracellular Ca$\^$2+/ concentration ([Ca$\^$2+/]$\_$i/) induced by glutamate. These data suggest ginsenosides Rb$_2$ and Rc prevent the cell swelling of astrocytes induced by glutamate, maybe via inhibition of Ca$\^$2+/ influx.

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

  • Im, Jeong-Hee;Kim, Song-Ja
    • Journal of Life Science
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    • v.20 no.4
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    • pp.519-527
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    • 2010
  • Paclitaxel is known as a potent inhibitor of microtubule depolymerization. It leads to mitotic arrest and cell death by stabilizing the spindle in various cell types. Here, we investigated the effects of paclitaxel on the proliferation and cell death of rabbit articular chondrocytes. Paclitaxel inhibited proliferation in a dose- and time- dependent manner, determined by MTT assay in rabbit articular chondrocytes. We also established paclitaxel-induced G2/M arrest by fluorescent activated cell sorter (FACS) analysis. Paclitaxel increased expression of cyclin B, p53 and p21, while reducing expression of cdc2 and cdc25C in chondrocytes, as detected by Western blot analysis. Interestingly, paclitaxel showed the mitotic catastrophe that leads to abnormal nucleus division and cell death without DNA fragmentation through activation of caspase. Cell death by mitotic catastrophe in cells treated with paclitaxel was suppressed by inhibiting G1/S arrest with 2 mM thymidine. These results demonstrate that paclitaxel induces cell death via mitotic catastrophe without activation of casepase in rabbit articular chondrocytes.

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

  • Ho Lee Chang;Beam Kwon Kang;Ho Jang Seung;Sun Song Yong;Gon Ryu Do
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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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.

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

  • Kim Hyeong Soo;Lee Yong Suk;Oh Suk Kyu;Lee Kang Chang;Lee Geon Mok;Lee Jeong;Lee Sang Bork;Kim Jong Ho;Yu Jun Ki;Kang Young Seong;Kim Sung Soo;Song Ho Jun;Park Seung Taeck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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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.

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 (비알코올성 지방간 세포 모델에 대한 택사, 산사, 구기자, 울금, 단삼, 인진의 효능 비교)

  • Han, Chang-Woo;Joo, Myung-Soo;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.33 no.4
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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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    • v.15 no.9
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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.

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

  • Kim, Jai Min;Park, Dong Hwa;Yoo, Geun Chang;Kim, Soon Ae;Cho, Seung Kwon
    • Journal of Korean Ophthalmic Optics Society
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    • v.7 no.2
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    • pp.95-99
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    • 2002
  • To investigate the manual procedure for custom fitting of ocular prosthesis and the cytotoxicity induced by extract solution of prosthetic eye, this study was performed. These procedures included the trial fitting of the wax model, impression molding, the an of iris color duplication, sclera mold made from the prosthetic wax model, scleral tints and vascular pattern, finishing and polishing. Inhibition of cell growth for extract solution was measured by MTT assay. Extract solution did not show the cytotoxicity. This study suggests that the manual procedure for custom fitting of ocular prosthesis is good for education of students.

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

  • Kim, Jai-Min;Lee, Seok-Ju;Seo, Eun-Sun
    • Journal of Korean Ophthalmic Optics Society
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    • v.7 no.2
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    • pp.189-195
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    • 2002
  • The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.

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