• Biomolecules & Therapeutics
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• v.19 no.2
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• pp.261-266
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• 2011

As recent reports suggest that nanoparticles may penetrate into cell membrane and effect DNA condition, it is necessary to assay possible cytotoxic and genotoxic risk. Three different sizes of magnetic nanoparticle silica (MNP@$SiO_2$) (50, 100 and 200 nm diameter) were tested for cytotoxicity and DNA damage using L5178Y cell. MNP@$SiO_2$ had constant physicochemical characteristics confirmed by transmission electron microscope, electron spin resonance spectrometer and inductively coupled plasma-atomic emission spectrometer for 48 h. Treatment of MNP@$SiO_2$ induced dose and time dependent cytotoxicity. At 6 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$ compared to vehicle control (p<0.05 or p<0.01). Moreover, at 24 h, 50 or 100 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$(p<0.01). And treatment of 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Furthermore, at 48 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Cellular location detected by confocal microscope represented they were existed in cytoplasm, mainly around cell membrane at 2 h after treatment of MNP@$SiO_2$. Treatment of 50 nm MNP@$SiO_2$ significantly increased DNA damage at middle and high dose (p<0.01), and treatment of 100 nm or 200 nm significantly increased DNA damage in all dose compared to control (p<0.01). Taken together, treatment of MNP@$SiO_2$ induced cytotoxicity and enhanced DNA damage in L5178Y cell.

• YAKHAK HOEJI
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• v.49 no.1
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• pp.30-37
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• 2005

Xuesaitong Ruanjiaonang (XR), a soft capsule containing Panax notoginseng saponins as main ingredients, is believed to remove extravasated blood and increase cerebral blood flow by improving blood circulation, and therefore, has been used in China to treat ischemic stroke or hemiplegia caused by cerebral thrombosis. To characterize pharmacological actions of XR, the present study evaluated its effects on neuronal cell damage induced by various oxidative insults or excitotoxic amino acids in primary cultured rat cortical cells. The neuronal cell viability was not affected by XR with the exposure for 2 h at the concentrations tested in this study ($10{\sim}1000\;{\mu}g/ml$). However, significant reduction of the cell viability was observed when the cultured cells were exposed to XR at $1000\;{\mu}g/ml$ for 24 h. XR was found to concentration-dependently inhibit the oxidative neuronal damage induced by $H_{2}O_2$, xanthine/xanthine oxidase or $Fe^{2+}$/ascorbic acid. In addition, it dramatically inhibited the excitotoxic damage induced by glutamate or N-methyl-D-aspartate (NMDA). We found that the NMDA-induced neurotoxicity was inhibited more effectively and potently than the glutamate-induced toxicity. Moreover, XR was found to exert mild inhibition of lipid peroxidation induced by $Fe^{2+}$/ascorbic acid in rat brain homogenates and some 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together, these results demonstrate neuroprotective and antioxidant effects of XR, showing inhibition of oxidative and excitotoxic damage in the cultured cortical neurons, as well as inhibition of lipid peroxidation and its radical scavenging activity. Considering that excitotoxicity and oxidative stress pl ay crucial roles in neuronal cell damage during ischemia and reperfusion, these results may provide pharmacological basis for its clinical usage to treat ischemic stroke.

• Journal of Dental Anesthesia and Pain Medicine
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• v.17 no.1
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• pp.37-46
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• 2017

Background: In oxidative stress, reactive oxygen species (ROS) production contributes to cellular dysfunction and initiates the apoptotic cascade. Autophagy is considered the mechanism that decreases ROS concentration and oxidative damage. Propofol shows antioxidant properties, but the mechanisms underlying the effect of propofol preconditioning (PPC) on oxidative injury remain unclear. Therefore, we investigated whether PPC protects against cell damage from hydrogen peroxide ($H_2O_2$)-induced oxidative stress and influences cellular autophagy. Method: COS-7 cells were randomly divided into the following groups: control, cells were incubated in normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) for 24 h without propofol; $H_2O_2$, cells were exposed to $H_2O_2$ ($400{\mu}M$) for 2 h; $PPC+H_2O_2$, cells pretreated with propofol were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)+PPC+H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide thiazolyl blue (MTT) reduction. Apoptosis was determined using Hoechst 33342 staining and fluorescence microscopy. The relationship between PPC and autophagy was detected using western blot analysis. Results: Cell viability decreased more significantly in the $H_2O_2$ group than in the control group, but it was improved by PPC ($100{\mu}M$). Pretreatment with propofol effectively decreased $H_2O_2$-induced COS-7 cell apoptosis. However, pretreatment with 3-MA inhibited the protective effect of propofol during apoptosis. Western blot analysis showed that the level of autophagy-related proteins was higher in the $PPC+H_2O_2$ group than that in the $H_2O_2$ group. Conclusion: PPC has a protective effect on $H_2O_2$-induced COS-7 cell apoptosis, which is mediated by autophagy activation.

• Nutrition Research and Practice
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• v.12 no.2
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• pp.93-100
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• 2018

BACKGROUND/OBJECTIVE: Oxidative stress plays a key role in neuronal cell damage, which is associated with neurodegenerative disease. The aim of present study was to investigate the neuroprotective effects of perilla oil (PO) and its active component, alpha-linolenic acid (ALA), against hydrogen peroxide $(H_2O_2)$-induced oxidative stress in SH-SY5Y neuronal cells. MATERIALS/METHODS: The SH-SY5Y human neuroblastoma cells exposed to $250{\mu}M$ $H_2O_2$ for 24 h were treated with different concentrations of PO (25, 125, 250 and $500{\mu}g/mL$) and its major fatty acid, ALA (1, 2.5, 5 and $25{\mu}g/mL$). We examined the effects of PO and ALA on $H_2O_2$-induced cell viability, lactate dehydrogenase (LDH) release, and nuclear condensation. Moreover, we determined whether PO and ALA regulated the apoptosis-related protein expressions, such as cleaved-poly ADP ribose polymerase (PARP), cleaved caspase-9 and -3, BCL-2 and BAX. RESULTS: Treatment of $H_2O_2$ resulted in decreased cell viability, increased LDH release, and increase in the nuclei condensation as indicated by Hoechst 33342 staining. However, PO and ALA treatment significantly attenuated the neuronal cell death, indicating that PO and ALA potently blocked the $H_2O_2$-induced neuronal apoptosis. Furthermore, cleaved-PARP, cleaved caspase-9 and -3 activations were significantly decreased in the presence of PO and ALA, and the $H_2O_2$-induced up-regulated BAX/BCL-2 ratio was blocked after treatment with PO and ALA. CONCLUSIONS: PO and its main fatty acid, ALA, exerted the protective activity from neuronal oxidative stress induced by $H_2O_2$. They regulated apoptotic pathway in neuronal cell death by alleviation of BAX/BCL-2 ratio, and down-regulation of cleaved-PARP and cleaved caspase-9 and -3. Although further studies are required to verify the protective mechanisms of PO and ALA from neuronal damage, PO and ALA are the promising agent against oxidative stress-induced apoptotic neuronal cell death.

• Biomolecules & Therapeutics
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• v.24 no.3
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• pp.338-345
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• 2016

Neurodegenerative diseases are often associated with oxidative damage in neuronal cells. This study was conducted to investigate the neuro-protective effect of methanolic (MeOH) extract of Perilla frutescens var. japonica and its one of the major compounds, rosmarinic acid, under oxidative stress induced by hydrogen peroxide ($H_2O_2$) in C6 glial cells. Exposure of C6 glial cells to $H_2O_2$ enhanced oxidative damage as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and thiobarbituric acid-reactive substance assays. The MeOH extract and rosmarinic acid prevented oxidative stress by increasing cell viability and inhibiting cellular lipid peroxidation. In addition, the MeOH extract and rosmarinic acid reduced $H_2O_2-indcued$ expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the transcriptional level. Moreover, iNOS and COX-2 protein expression was down-regulated in $H_2O_2-indcued$ C6 glial cells treated with the MeOH extract and rosmarinic acid. These findings suggest that P. frutescens var. japonica and rosmarinic acid could prevent the progression of neurodegenerative diseases through attenuation of neuronal oxidative stress.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.6
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• pp.857-866
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• 2011

This study examined the metabolism of free radicals in hepatic mitochondria of goats induced by lipopolysaccharide (LPS), and investigated the effects of melatonin (MT). Forty-eight healthy goats ($10{\pm}1.2\;kg$) were randomly selected and divided into four groups: saline control, LPS, MT+LPS and MT. The goats within each group were3 sacrificed either 3 or 6 h after treatment and the livers removed to isolate mitochondria. The respiration control ratio (RCR), the ADP:O ratio, the oxidative phosphorylation ratio (OPR), the concentration of $H_2O_2$ and the activities of Complex I-IV were determined. The mitochondrial membrane potential ($\Delta\psi_m$) was analyzed by flow cytometry. The results showed that RCR, O/P and OPR of the LPS group decreased (p<0.05), as well as activities of respiratory complexes, whereas the generation of $H_2O_2$ in Complex III increased (p<0.05) after 3 h, while Complex II and III increased after 6 h. Also, it was found that the mitochondrial membrane potential of the LPS group declined (p<0.05). However, pre-treatment with MT attenuated the injury induced by LPS, which not only presented higher (p<0.05) RCR, O/P, OPR, and respiratory complex activities, but also maintained the $\Delta\psi_m$. Interestingly, it is revealed that, in the MT+LPS group, the generation of $H_2O_2$ increased firstly in 3 h, and then significantly (p<0.05).decreased after 6 h. In the MT group, the function of mitochondria, the transmenbrane potential and the generation of $H_2O_2$ were obviously improved compared to the control group. Conclusion: melatonin prevents damage caused by LPS on hepatic mitochondria of goats.

• Journal of Plant Biotechnology
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• v.41 no.2
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• pp.73-80
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• 2014

Pretreatment of chinese cabbage leaves with $100{\mu}M$ sodium nitroprusside (SNP), a nitric oxide (NO) donor, effectively improved their tolerance to subsequent $2{\mu}M$ paraquat (PQ)-induced oxidative damage. The fresh weight, and chlorophyll and protein contents in primary leaves treated with PQ alone were noticeably reduced over 24 h light incubation. However, these leaf injury symptoms were significantly alleviated with $100{\mu}M$ SNP pretreatment for 3 h prior to PQ exposure. In additions, the increase of the contents of malondialdehyde (MDA) and $H_2O_2$ due to PQ exposure were significantly inhibited by SNP pretreatment. Together with the protective effects of SNP against PQ toxicity in leaves, the changes of ascorbate-glutathione cycle enzymes activities were examined. In the PQ alone treatment, the activities of APX, DHAR, and GR after 6 h incubation were rapidly reduced and showed 19%, 50% and 39% respectively, compared with those of the control. However, the decreases in these enzyme activities were significantly inhibited by SNP pretreatment. As a result, their activities were higher than those of PQ alone treatment by 5 times, 2 times, and 1.5 times, respectively, at 6 h incubation. Thereafter, these enzymes decrease their activities gradually showing high levels than those of PQ alone. Based on the above results, it can be assumed that the activation of ascorbate-glutathione cycle by SNP pretreatment in chinese cabbage leaves exposed to PQ can prevent $H_2O_2$ accumulation, thereby leading to protection against PQ-induced oxidative stress. Also, these results indicate that NO acts as an protectant against PQ stress in the leaves of chinese cabbage.

• Korean Journal of Acupuncture
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• v.24 no.1
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• pp.171-187
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• 2007

Objectives : This study was performed to determine if Orostachys japonicus A. Berger herbal acupuncture (OjB) provides the protective effect against the loss of cell viability and DNA damage induced by oxidant in renal proximal tubular cells. Methods : The cell viability was evaluated by a MTT reduction assay and DNA damage was estimated by measuring double stranded DNA breaks in opossum kidney (OK) cells, an established proximal tubular cell line. Lipid peroxidation was determined by measuring malondialdehyde (MDA), a product of lipid peroxidation. Results : H2O2 increased the loss of cell viability in a time-dependent manner, which were prevented by 0.1% OjB. The protective effect of OjB was dose-dependent over concentration range of 0.05-0.5%. H2O2 caused ATP depletion and DNA damage, which were prevented by OjB and the hydrogen peroxide scavenger catalase. The loss of cell viability by H2O2 was not affected by the antioxidant DPPD, but lipid peroxidation by the oxidant was completely inhibited by DPPD. Generation of superoxide and H2O2 in neutrophils activated by phorbol-12,13-dibutyrate was inhibited by OjB in a dose-dependent manner. OjB inhibited generation of H2O2 in OK cells treated with antimycin A and exerted a direct H2O2 scavenging effect. Exposure of OK cells to 1 mM tBHP caused a significant depletion of glutathione which was prevented by OjB. OjB accelerated the recovery in cells cultured for 20 hr in normal medium without oxidant following oxidative stress. Conclusions : These results suggest that OjB exerts the protective effect against oxidant-induced cell injury and its protective effect was resulted from radical scavenging and antioxidant activities.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.473-479
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• 2020

Endothelial cell injury is a major contributor to cardiovascular diseases. The 2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-Glucoside (TSG) contributes to alleviate human umbilical vein endothelial cells (HUVECs) injury through mechanisms still know a little. This study aims to clarify the TSG effects on gene expression (mRNA and microRNA) related to oxidative stress and endoplasmic reticulum stress induced by H₂O₂ in HUVECs. We found that TSG significantly reduced the death rate of cells and increased intracellular superoxide dismutase activity. At qRT-PCR, experimental data showed that TSG significantly counteracted the expressions of miR-9-5p, miR-16, miR-21, miR-29b, miR-145-5p, and miR-204-5p. Besides, TSG prevented the expression of ATF6 and CHOP increasing. In contrast, TSG promoted the expression of E2F1. In conclusion, our results point to the obvious protective effect of TSG on HUVECs injury induced by H₂O₂, and the mechanism may through miR16/ATF6/ E2F1 signaling pathway.

• Korean Journal of Medicinal Crop Science
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• v.25 no.5
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• pp.315-321
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• 2017

Background: Glycyrrhiza uralensis Radix (GR) is a crude drugs used in Asian countries that has been reported to prevent the progression of neurodegenerative diseases such as Alzheimer's disease. The present study examined whether GR and its active compounds, glycyrrhizic acid (GA) and isoliquiritigenin (IL), exerted protective effects on $H_2O_2$-induced oxidative damage in C6 glial cells. Methods and Results: We exposed C6 glial cells to hydrogen peroxide ($H_2O_2$) for 24 h and investigated the cellular response to GR and its active compounds by evaluating cell viability, reactivie oxygen species (ROS) production, and apoptosis-related protein expression. GR successfully mitigated the reduced cell viability and ROS production induced by $H_2O_2$ in C6 glial cells, IL and GA significantly increased the cell viability and decreased ROS production. In addition, IL and GA down-regulated apoptotic Baxdependent caspase-3 activation, but each compound exerted different mechanisms, i.e., IL dose-dependently decreased ROS production and, GA up-regulated anti-apoptotic Bcl-2 expression. Conclusions: These results demonstrated that GR and its active components, IL and GA, exhibit potential for use as natural neurodegenerative agents for the modulation of apoptosis in C6 glial cells.