• Journal of Microbiology and Biotechnology
• /
• v.14 no.2
• /
• pp.350-355
• /
• 2004

Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

• Journal of Oriental Neuropsychiatry
• /
• v.11 no.1
• /
• pp.1-18
• /
• 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.

• Herbal Formula Science
• /
• v.8 no.1
• /
• pp.319-328
• /
• 2000

To evaluate the effect of Baepungtang(BPT) water extract on cultured mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MTT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of BPT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cell on NR assay and MTT assay. 2. $MTT_{50}$ value and $NR_{50}$ value of XO/HX were 30 mU/ml XO/O.2 mM HX. 3. BPT water extract have efficacy of increasing neurofilament. 4. BPT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that BPT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.16 no.3
• /
• pp.563-566
• /
• 2002

To evaluate the effect of Jingansikpung-tang(JST) water extract on cultured mouse spinal sensory neuron which was inhibited by glucose oxidase(GO)-induced cytotoxicity, MTT and LDH activity assay were carried out after the cultured mouse spinal sensory neuron were pre-incubated with various concentrations of JST extract for 3 hours prior to exposure of GO. The results obtained were as follows: GO, a oxygen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cells on MTT assay. JST water extract have efficacy of decreasing LDH activity increasing by GO in cultured mouse spinal sensory neuron. From above the results, it is concluded that JST water extract has marked efficacy as a treatment for the damages caused in the GO-mediated oxidative process.

• Environmental Mutagens and Carcinogens
• /
• v.18 no.1
• /
• pp.37-42
• /
• 1998

In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

• The Journal of Korea CHUNA Manual Medicine
• /
• v.2 no.1
• /
• pp.143-157
• /
• 2001

Objectives and Methods : To evaluate the mechanism of oxidative damage by xanthine oxydase(XO) and hypoxanthine(HX)-induced oxygen radicals, MTT assay and NR assay were carried out after the cultured mouse spinal sensory neurons were preincubated for 4 hours with various concentrations of XO/HX. And the amount of total protein. neurofilament EIA. lipid peroxidation and LDH activity were measured, to evaluate the protective effect of Herbar Chelidonii(HC) water extract on cultured spinal sensory neurons damaged by XO/HX. after the cultured mouse spinal sensory neurons were preincubated with various concentrations of HC water extract for 3 hours prior to exposure of XO/HX. Results : XO/HX decreased significantly the survival rate of the cultured mouse sensory neurons by NR assay and MTT assay In proportion to concentration and exposed time. In proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA. the decrease of total protein amount by SRB assay and the Increase of lipid peroxidation as well as LDH. HC showed the quantitative increase of neurofilament and total protein, but showed the decrease of lipid peroxidation and LDH activity against the neurotoxicity of XO/HX. Conclusions : From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and that the herbs extract, such as HC, prevent the toxicity of XO/HX effectively in that they decrease lipid peroxidation and LDH activity.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.1
• /
• pp.115-119
• /
• 2008

This study was done to evaluate the antioxidant effect of Crataegi Fructus (CF) extract on the oxidative stress induced by reactive oxygen species (ROS), The human skin fibroblasts (Detroit 551) were cultured with various concentrations of hydrogen peroxide $(H_2O_2)$. The cytotoxicity of $H_2O_2-induced$ oxidative stress was performed by XTT assay for the cell viability according to the dose- and time-dependent treatment. For the protective effect of CF extract on $H_2O_2-mediated$ oxidative stress, cell viability, lactate dehydroganase (LDH) activity, and ferric thiocyanate (FTC) assay for the inhibitive activity of lipid peroxidation on CF extract were carried out. In this study, $H_2O_2-mediated$ oxidative stress was decreased cell viability dose-, and time-dependent manner and increased LDH activity compared with the control in these cultures. In the protective effect, CF extract increased cell viability and decreased LDH activity on $H_2O_2-mediated$ oxidative stress, especially, CF extract has antioxidant effect by the showing the inhibitive activity of lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2-mediated$ oxidative stress was highly toxic, and also, CF extract showed the protective effect on $H_2O_2-mediated$ oxidative stress by showing the increased cell viability, decreased LDH activity and lipid peroxidation inhibition in these cultures.

• Ceramist
• /
• v.22 no.3
• /
• pp.301-315
• /
• 2019

We prepared hybrids between layered double hydroxide (LDH) and natural plant extract such as Peaonia suffruticosa Andrews (PS) and Peaonia Japonica (PJ) which was confirmed anti-bacterial activity through paper disc diffusion assay. According to X-ray diffractometer, scanning electron microscope, zeta-potential measurement and quantification of extract loading amount in hybrids, we confirmed that similar amount of PS and PJ loaded on inter-particle pore of LDH with partial adsorption on surface of LDH through reconstruction process. We also evaluated the bacterial colony forming inhibition of PS extract, PJ extract, PS-LDH and PJ-LDH hybrids against Escherichia coli as gram negative bacterium and Bacillus subtilis as gram positive bacterium, suggesting that both hybrids have enhanced anti-bacterial activity compared with extract itself.

• Journal of Sasang Constitutional Medicine
• /
• v.13 no.1
• /
• pp.88-96
• /
• 2001

To evaluate the effect of Yuldahansotang(YHT) water extract on cultuted mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MIT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried our after the cultured mouse spinal sensory neuron were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cells on NR assay and MTT assay. 2. MTT50 value and NR50 value pf XO/HX were 20 mU/ml XO/0.2 mM HX and 40 mU/ml XO/0.2 mM HX. 3. YHT water extract have efficacy of increasing neurofilament. 4. YHT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that YHT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

• Journal of Sasang Constitutional Medicine
• /
• v.13 no.1
• /
• pp.70-78
• /
• 2001

ADR유발성 심근독성에 대한 심근세포의 손상기전을 규명하기 위해 ADR의 독성을 MTT정량, NR정량, LDH활성도 및 심박동을 측정하였다. 배양된 심근세포에서 청심연자탕 전탕액의 심근세포 보호효과는 LDH활성도 측정과 심박동 측정을 통해 관찰할 수 있었다. 이 실험을 통해 다음과 같은 결과를 얻을 수 있었다. 1. ADR은 배양심근세포에서 세포의 생존능력을 떨어뜨렸고, LDH의 활성도를 높였으며, 심박동수를 감소시켰다. 2. 청심연자탕 전탕액은 배양심근세포에서 ADR에 의해 증가된 LDH 활성도를 유의하게 감소시켰다. 3. 청심연자탕 전탕액은 배양심근세포에서 ADR에 의해 감소된 심박동을 유의하게 증가시켰다. 이상의 결과를 통해 ADR은 신생 마우스에서 적출해낸 배양 심근세포에서 독성효과를 나타냈음을 알 수 있었으며, 청심연자탕 전탕액은 ADR에 의해 유발된 심근세포독성에 매우 효과적으로 방어효과를 나타냄을 알 수 있었다.