• Korean Journal of Clinical Laboratory Science
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• v.51 no.2
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• pp.205-213
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• 2019

The protective effect of Agrimonia pilosa var. (AP) extract on potassium dichromate ($K_2Cr_2O_7$)-induced cytotoxicity in cultured NIH3T3 fibroblasts, was examined by performing an XTT assay for the cell viability and antioxidative effects, such as lactate dehydrogenase (LDH) activity and superoxide anion-radical (SAR) scavenging activity. In this study, $K_2Cr_2O_7$ decreased the cell viability significantly in a dose-dependent manner, and the $XTT_{50}$ value was determined to be $37.5{\mu}M$, which was highly-toxic according to the Borenfreund and Puerner' toxic criteria. The antioxidant, butylated hydroxytoluene (BHT), increased remarkably the cell viability damaged by $K_2Cr_2O_7$-induced cytotoxicity in these cultures. With regard to the protective effect of the AP extract on $K_2Cr_2O_7$-induced cytotoxicity, AP extract produced a significant increase in cell viability and antioxidative effects as the inhibitory ability LDH and SAR scavenging ability. These findings suggest that oxidative stress is involved in the cytotoxicity of $K_2Cr_2O_7$, and the AP extract effectively protected the cells from $K_2Cr_2O_7$-induced cytotoxicity by antioxidative effects. These results suggest that natural resources, such as AP extract, may be a putative therapeutic agent for the diminution or treatment of cytotoxicity induced by heavy metallic bases, such as $K_2Cr_2O_7$ correlated with oxidative stress.

• Archives of Pharmacal Research
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• v.25 no.4
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• pp.469-474
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• 2002

The inner shell of the chestnut (Castanea crenata S. et Z., Fagaceae) has been used as an anti-wrinkle/skin firming agent in East Asia, and preliminary experiments have found that a 70% ethanol extract from this plant material can prevent cell detachment of skin fibroblasts from culture plates. In order to examine the molecular mechanisms underlying this phenomenon, its effects on the expression of adhesion molecules, such as fibronectin and vitronectin, were investigated using the mouse skin fibroblast cell line, NIH/3T3. Using fixed-cell ELISA, Western blotting and immunofluorescence cell staining, it was clearly demonstrated that the chestnut inner shell extract enhanced the expression of the cell-associated fibronectin and vitronectin. Scoparone (6,7-dimethoxycoumarin), isolated from the extract, also possessed similar properties. These findings suggest that the enhanced expression of the adhesion molecules may be one of the molecular mechanisms for how the chestnut inner shell extract preventing cell detachment and may be also responsible for its anti-wrinkle/skin firming effect.

• BMB Reports
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• v.49 no.2
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• pp.99-104
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• 2016

The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

• BMB Reports
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• v.35 no.5
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• pp.452-458
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• 2002

Exposure to ultraviolet light (UV) induces apoptosis in mammalian cells, The caspase group of proteases is required for the appotosis. This pathway is initiated by a release of cytochrome c from the mitochondria into the cytosol. Several Bcl-2 family proteins can regulate the release of cytochrome c by stabilizing the mitochondrial membrane. Here we show that expression of the endogenous bcl-xL was strongly downregulated in NIH3T3 cells within 2 h after UV-C irradiation, and that of bax was upregulated from 8 h after irradiation. Apoptosis was induced in more than 50% of the NIH3T3 cells 48 h after irradiation. Constitutive overexpression of bcl-xL in NIH3T3 cells protected the UV-induced apoptosis by preventing the loss of mitochondrial membrane potential and the activation of caspase 9. There results suggest that downregulation of Bcl-xL is relevant to UV-induced apoptosis of tibroblasts.

• 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.

• 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.

• Macromolecular Research
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• v.10 no.3
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• pp.150-157
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• 2002

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a microbial storage polymer with biodegradable properties. In order to improve the cell compatibility of PHBV surfaces, the physicochemical treatments have been demonstrated. In this study, physical method was corona discharge treatment and chemical method was chloric acid mixture solution treatment. The physicochemically treated PHBV film surfaces were characterized by the measurement of water contact angle, electron spectroscopy for chemical analysis, and scanning electron microscopy (SEM). The water contact angle of the physicochemically treated PHBV surfaces decreased from 75 to 30～40 degree, increased hydrophilicity. due to the introduction of oxygen-based functional group onto the PHBV backbone chain. The mouse NIH/3T3 fibroblasts cultured onto the physicochemically treated PHBV film surfaces with different wettability. The effect of the PHBV surface with different wettability was determined by SEM as counts of cell number and [$^3$H]thymidine incorporation as measures of cell proliferation. As the surface wettability increased, the number of the cell adhered and proliferated on the surface was increased. The result seems closely related with the serum protein adsorption on the physicochemically treated PHBV surface. In conclusion, this study demonstrated that the surface wettabilily of biodegradable polymer as the PHBV plays an important role for cell adhesion and proliferation behavior for biomedical application.

• Restorative Dentistry and Endodontics
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• v.29 no.4
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• pp.370-377
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• 2004

The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 ($1{\;}{\times}{\;}10^5{\;}cells/ml/well$) were cultured at 12-well plate at $37^{\circ}C$ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05), 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

• Korean Journal of Clinical Laboratory Science
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• v.42 no.3
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• pp.116-124
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• 2010

This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

• Biomedical Science Letters
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• v.24 no.4
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• pp.357-364
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• 2018

Manganese (Mn) is used as main materials in various chemical processes of industry, but it suggested that Mn brings about its toxicant by fume or dust through respiratory system and skin barrier. Mn toxicant induces the loss of mental health and life quality by cerebrovascular and skin diseases. Nevertheless, it lefts much unknown on the mechanism and the effectively therapeutic methods about Mn toxicant. Therefore, this study was evaluated the cytotoxicity induced by manganese dioxide ($MnO_2$) in cultured NIH3T3 fibroblasts, and also, the correlation between $MnO_2$-induced cytotoxicity and oxidative stress was examined. While, the effect of Eclipta prostrata L. (EP) extract belong to Compositae was assessed against $MnO_2$-induced cytotoxicity in the view of antioxidative effect for searching the natural resources mitigating or preventing the $MnO_2$-induced cytotoxicity. In this study, $MnO_2$-induced cytotoxicity was revealed as mid-toxic by Borenfreud and Puerner's toxic criteria, and catalase (CAT), an antioxidant prevented $MnO_2$-induced cytotoxicity by the remarkable increase of cell viability in these cultures. While, in the protective effect of EP extract on $MnO_2$-induced cytotoxicity, EP extract effectively prevented the cytotoxicity induced by $MnO_2$ via antioxidative effects such as xanthine oxidase (XO) inhibitory ability and DPPH-radical scavenging ability. From the above results, EP extract showed the effective prevention against $MnO_2$-induced cytotoxicity correlated with oxidative stress by antioxidative effects. Conclusively, this study may be useful to research or development the alternatively therapeutic agent from natural resources like EP extract for the treatment of diseases resulted in oxidative stress.