• Title/Summary/Keyword: MTT and SRB assays

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A Study on the Effects of Ramulus et Uncus Uncariae (REUU) on the Cultured Spinal Dorsal Root Ganglion Neurons Damaged by Oxygen Free Radicals (조구등(釣鉤藤)이 산소자유기(酸素自由基)에 의하여 손상(損傷)된 배영척수감각신경절세포(培養脊髓感覺神經節細胞)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Kang, Hyung-Won;Park, Jin-Sung
    • Journal of Oriental Neuropsychiatry
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    • v.11 no.1
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    • pp.1-18
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    • 2000
  • To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide $(H_{2}O_{2})$ on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice, Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of $H_{2}O_{2}$ for 5 hours, after which the neurotoxic effect of $H_{2}O_{2}$ was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows. 1. In NR assay and MTT assay, $H_{2}O_{2}$ significantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures. 2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of $H_{2}O_{2}$ showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity. 3. The effect of Ramulus et Uncus Uncariae (REUU) on $H_{2}O_{2}$ induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity. These results suggest that $H_{2}O_{2}$ has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting $H_{2}O_{2}$ induced neurotoxicity by decreasing lipid peroxidation and LDH activity.

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The Inhibitory Effects of Trichosanthes kirilowii Root against Cadmium Induced Cytotoxicity (III) (천화분의 카드뮴에 대한 독성억제효과(III))

  • Lee, Jeong-Ho;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Chung, Woo-Young;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.15-21
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    • 2001
  • This study was conducted to investigate the antitoxic components in the water extract of the roots of Trichosanthes kirilowii (Cucurbitaceae). The results were as follows: Generally, detoxication effects by the water extract of T. kirilowii increased in proportion to the concentrations. Experimental animals were treated with cadmium and T. kirilowii water extract by oral administration. When 40 mg/kg dosage of T. kirilowii extract was administrated it showed the highest antitoxic effects in metallothionein induction. After the water extract treatment, body weights did not increase in proportion to the extract concentrations. These results suggest that T. kirilowii extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of water extract of T. kirilowii on NIH 3T3 fibroblasts was evaluated by the 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. These results were obtained as follows; The concentration of $10^{-2}\;mg/ml$ of T. kirilowii extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that T. kirilowii extract retains a potential antitoxic activity.

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The Inhibitory Effects of the Methanol Extract of Houttuynia cordata $T_{HUNB}$ against Cadmium Induced Cytotoxicity (V) (어성초 메탄올 추출물의 카드뮴에 대한 독성억제효과(V))

  • Lee, Jeong-Ho;Jeong, Seung-Il;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.61-67
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    • 2001
  • This study was conducted to investigate the antitoxic agent in methanol extract of Houttuynia cordata $T_{HUNB}$. Detoxication effects By H. cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations. When 40 mg/kg dosage of H. cordata $T_{HUNB}$ extract was administered, it showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights generally increased in proportion to the extract concentrations. From the above results, H. cordata $T_{HUNB}$ extract increased metallothionein concentrations and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of methanol extract of H. cordata $T_{HUNB}$ on NIH 3T3 fibroblasts was evaluated by the MTT {3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide} and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells, $10^{-2}\;mg/ml$ Concentrations of H. cordata $T_{HUNB}$ extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that H. cordata $T_{HUNB}$ extract retains a potential antitoxic activity.

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Regenerative Effects of Houttuynia cordata Extracts against Cadmium-induced Cytotoxicity in NIH 3T3 fibroblasts Cultures (VII) (카드뮴 독성으로 손상된 NIH 3T3 섬유모세포에 대한 어성초 추출물의 재생효과(VII))

  • Lee Jeong-Ho;Oh Hyun-Ju;Jeong Seung-Il;Chun Hyun-Ja;Han Do-Seok;Lee Hyun-Ok;Ahn Jong-Woong;Lee Mee-Hee;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.5 no.2
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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.

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Antitumor evaluation and antimicrobial activity of geranyl phenyl ethers

  • Oh Hyun-Ju;Shin Ji-Hee;Chung Woo-Young;Han Du-Seok;Baek Seung-Hwa
    • Advances in Traditional Medicine
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    • v.2 no.1
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    • pp.52-57
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    • 2002
  • Geranyl phenyl ethers 1, 2, 3, 4 and 5, 5-fluorouracil 6 and adriamycin 7 as references were tested for their growth inhibitory effects against tumor cell lines and NIH 3T3 fibroblasts using two different assays, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulforhodamine B protein (SRB) assays. These results suggest that methyl-4-[{(2E)-3,7-dimethyl-2,6-octadienyl}oxy]-3-methoxy benzoate 1 showed growth inhibition activity against tumor cell lines. The maximum activity exhibited by methyl-4-[{(2E)-3,7-dimethyl-2,6-octadienyl}oxy]-3-hydroxybenzoate 3 against Staphylococcus epidermidis (MIC, $1,000{\mu}g/ml$).

Orthosiphon pallidus, a Potential Treatment for Patients with Breast Cancer

  • Singh, Mukesh K.;Dhongade, Hemant;Tripathi, Dulal Krishna
    • Journal of Pharmacopuncture
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    • v.20 no.4
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    • pp.265-273
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    • 2017
  • Objective: Orthosiphon pallidus (O. pallidus), which belongs to the Lamiaceae family, is a popular garden plant that is widely used for the treatment of various diseases, such as urinary lithiasis, fever, hepatitis, cancer and jaundice. The objective of the present work was to investigate the antioxidant free-radical scavenging and the anticancer activities of O. pallidus against human breast-cancer cell lines. Methods: The antioxidant activity of Orthosiphon pallidus aqueous extract (OPAE) was investigated using different models, such as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the 2, 2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) models, as were the $Fe^+$ chelation, the hydroxyl radical and superoxide radical scavenging, and total reducing power activities. The anticancer activities of the extract were determined by using the 3-(4, 5- dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) and the sulforhodamine (SRB) assays on the MCF-7 and the MDA-MB-231 cancer cell lines. Results: The aqueous Orthosiphon pallidus extract showed potent activity in in-vitro models. It significantly inhibited the scavenging of hydroxyl and superoxide radicals, but induced a remarkable $Fe^+$ chelation activity. For both cell lines, the percent cytotoxicity was found to increase steadily with increasing OPAE concentration up to $240{\mu}g/mL$. Conclusion: These results suggest that Orthosiphon pallidus has excellent antioxidant, antimicrobial, and anticancer activities against human breast-cancer cell lines.

In vitro cytotoxicity and in vivo acute toxicity of selected polysaccharide hydrogels as pharmaceutical excipients

  • Kulkarni GT;Gowthanarajan K;Raghu C;Ashok G;Vijayan P
    • Advances in Traditional Medicine
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    • v.5 no.1
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    • pp.29-36
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    • 2005
  • Polysaccharide hydrogels constitute a structurally diverse class of biological macromolecules with a wide range of physicochemical properties. They also constitute important members of the family of industrial water-soluble polymers. They find application in Pharmacy as binders, disintegrants, suspending, emulsifying and sustaining agents. According to the International Pharmaceutical Excipients Council (IPEC), an excipient must have an established safety profile. Hence, in the present study, in vitro cytotoxicity on Vero and HEp-2 cell lines, and in vivo acute toxicity in rats were carried out to establish the safety of polysaccharide hydrogels from the seeds of Plantago ovata and Ocimum basilicum. The in vitro cytotoxicity was determined by MTT and SRB assays. In the in vivo acute toxicity, the effects of three different doses of hydrogels (100, 200 and 400 mg/kg body weight) on food and water intake, body weight, biochemical and hematological parameters were studied. The results of in vitro did not show any cytotoxicity on both the cell lines used. In the in vivo acute toxicity, the hydrogels did not show any toxic symptoms in all three dose levels. This establishes the safety of the selected hydrogels. Hence, they can be used as excipients in pharmaceutical dosage forms.

In Vitro Anticancer Activities of Anogeissus latifolia, Terminalia bellerica, Acacia catechu and Moringa oleiferna Indian Plants

  • Diab, Kawthar AE;Guru, Santosh Kumar;Bhushan, Shashi;Saxena, Ajit K
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6423-6428
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    • 2015
  • The present study was designed to evaluate in vitro anti-proliferative potential of extracts from four Indian medicinal plants, namely Anogeissus latifolia, Terminalia bellerica, Acacia catechu and Moringa oleiferna. Their cytotoxicity was tested in nine human cancer cell lines, including cancers of lung (A549), prostate (PC-3), breast (T47D and MCF-7), colon (HCT-16 and Colo-205) and leukemia (THP-1, HL-60 and K562) by using SRB and MTT assays. The findings showed that the selected plant extracts inhibited the cell proliferation of nine human cancer cell lines in a concentration dependent manner. The extracts inhibited cell viability of leukemia HL-60 and K562 cells by blocking G0/G1 phase of the cell cycle. Interestingly, A. catechu extract at $100{\mu}g/mL$ induced G2/M arrest in K562 cells. DNA fragmentation analysis displayed the appearance of a smear pattern of cell necrosis upon agarose gel electrophoresis after incubation of HL-60 cells with these extracts for 24h.

Cytotoxicity and Antigenotoxic Effects of Cordyceps militaris Extracts (번데기동충하조(Cordyceps militaris) 추출물의 세포독성 및 유전독성 억제효과)

  • 김미남;최승필;이득식;함승시
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.921-927
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    • 2001
  • This study was performed to investigated the effects on the cytotoxicity and antigenotoxicity of Cordyceps militaris extracts on the human cancer cell lines. The ethanol extract and five fractions which were hexane, chloroform, ethylacetate, butanol and aqueous were screened for crytotoxicity on human lung carcinoma(A549). human breast adenocarcinoma (MCF-7) human epitheloid carcinoma(HeLa), human fibrosarcoma(HT1080) human hepatocellular carcinoma(Hep3B), human gastric carcinoma(KATOIII) and chronic myelogenous leukemia(K562) cell by SRB and MTT assays. The results showed that growth inhibition rates of the human cancer cell in the presence of Cordyceps militaris were inhibited with increasing concentration of the extract. The ethanol extract from Cordyceps militaris had strong inhibitory effects in1 mg/mL treatment by SRB assay , showing 89.4%, 85.7%, 72.9% and 65.5% inhibition in HT1080, HeLa, Hep3B and A549, respectively. The treatment of 1 mg/mL hexane fraction by SRB assay had the strongest cytotoxicity with 97.0% on HT1080 followed by MCF-7(92.9%) and HeLA(90.3%). The inhibition ration on KATOIII by MTT assay was much higher in the butanol (83.7%) and aqueous (80.4%) than in the ethanol extract (61.5%) And also, K562 showed similar tendency with KATOIII. The effects of Cordyceps militaris extracts on the frequencies of micronucleated polychromatic erythrocytes (MNPCEs) induced by N-methyl-N-nitro-N-nitrosoguanidime(MNNG) were investigated in the bone-marrow cells of ICR male mice. The amount of 10, 20, 40 and 80 mg/kg of each extract were administered to animals immediately after injection of MNNG, and the exposure time was 36 hours. Significant reductions(p<0.05) with 39.7%, 52.7%, 71.4% and 83.9% were observed in the frequencies of MNPCE when 10, 20, 40 and 80 mg/kg of the hexane fraction of Coryceps militarus extracts were given to the mice.

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Comparative Antitumor Activity of Different Solvent Fractions from an Auricularia auricula-judae Ethanol Extract in P388D1 and Sarcoma 180 Cells

  • Reza, Ahsanur;Choi, Myung-Jin;Damte, Dereje;Jo, Woo-Sik;Lee, Seung-Jin;Lee, Joong-Su;Park, Seung-Chun
    • Toxicological Research
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    • v.27 no.2
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    • pp.77-83
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    • 2011
  • The objective of this study was to evaluate and compare the antitumor activity of different solvent fractions (ethanol, dichloromethane, ethyl acetate, butanol and water) of the Auricularia auricula-judae 70% ethanol extract on the P388D1 macrophage and sarcoma 180 cells. A dose-dependent antitumor activity of each solvent fraction (from 0.01 mg/ml to 0.3 mg/ml) was shown against both cell types. These cytotoxic effects of all the tested fractions were confirmed on the MTT and SRB assays, without statistical differences each other. $IC_{50}$ value of dichloromethane fraction was 94.2 ${\mu}g/ml$ against sarcoma 180 cells lower than any other solvent fractions. The potent antitumor effect of the dichloromethane (DCM) fraction was also found against solid tumor in BALB/c mice. The splenomegaly and higher splenic index were found in tumor-bearing mice, with the DCM fraction returning to the negative control values. Thus, the results indicated the dichloromethane fraction may have potential ingredients as antitumor candidates.