Neuroprotective effects of N-acetylglucosamine against hydrogen peroxide-induced apoptosis in human neuronal SK-N-SH cells by inhibiting the activation of caspase-3, PARP, and p38

Neuroprotective effects of N-acetylglucosamine (GlcNAc), a monosaccharide derivative of glucose, against H₂O₂-induced neurotoxicity and its underlying mechanism in human SK-N-SH neuroblastoma cells were investigated. Pretreatment of GlcNAc prior to exposure of cells to H₂O₂ stress significantly reduced the H₂O₂-mediated neuronal cell death and apoptosis. The GlcNAc dose-dependently decreased the level of intracellular reactive oxygen species (ROS) in H₂O₂-treated cells and also effectively inhibited H₂O₂-induced apoptotic features such as DNA fragmentation, caspase-3, and poly ADP-ribose polymerase (PARP) cleavages, and p38 phosphorylation. These results suggested that GlcNAc might potentially serve as agents for prevention of neurodegenerative diseases caused by oxidative stresses and this effect may be associated with the suppression of caspase-3, PARP, and p38 activation.     

Molecular mechanism underlying anti-apoptotic and anti-inflammatory effects of Mamao (Antidesma thwaitesianum Müll. Arg.) polyphenolics in human breast epithelial cells

There has been increasing interest in finding natural antioxidants to prevent free radical damage and retard the progress of chronic inflammatory diseases. Our previous data demonstrated the strong antioxidant properties of polyphenolics in Mamao seed (MS) and Mamao marc (MM) extracts. In this study we further investigated the effect of MS and MM polyphenolics on hydrogen peroxide (H₂O₂)-induced apoptosis and tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation, using human breast epithelial (MCF10A) cells. MS and MM extracts conferred dose-dependent protection against H₂O₂-induced apoptosis by inhibiting PARP/caspase-3 cleavage, inducing anti-apoptotic Bcl-2 expression, and down-regulating pro-apoptotic Bax. Moreover, MS and MM polyphenolics inhibited TPA-induced COX-2 and NF-κB activation by blocking the degradation of cytoplasmic IκBα, as well as subsequent nuclear translocation of p65 and attenuation of the activation of ERK, but not JNK and p38. These data establish the molecular mechanism for the anti-apoptotic and anti-inflammatory effects of MS and MM polyphenolics.     

The component of green tea,L-theanine protects human hepatic L02 cells from hydrogen peroxide-induced apoptosis

L-theanine is a natural amino acid in green tea and it has been well known for its activities of relieving depression and neuroprotection. However, cytoprotective effect and its mechanism of L-theanine on hepatocytes have not been reported. The objective of this work was to investigate the hepatoprotective effect of L-theanine as well as its mechanism by using the human hepatic L02 cells injured by hydrogen peroxide (H₂O₂). Results showed that L-theanine dose dependently decreased H₂O₂-induced cell viability loss and lactate dehydrogenase (LDH) release. L-theanine pretreatment improved nuclear morphology of the cells injured by H₂O₂. By using flow cytometric analysis, we found that L-theanine significantly inhibited H₂O₂-induced cell apoptosis. Further, L-theanine attenuated H₂O₂-induced reduction in pro-caspase3 and cleavage of poly (adenosine diphosphate-ribose) polymerase (PARP). H₂O₂-activated p38 mitogen-activated protein kinase (MAPK) was also inhibited by L-theanine. These data suggest that L-theanine could protect L02 cells against H₂O₂-induced apoptosis via suppression of p38 MAPK. L-theanine might potentially be useful in the prevention and treatment of liver diseases.     

Antiproliferative effects of cherry juice and wine in Chinese hamster lung fibroblast cells and their phenolic constituents and antioxidant activities

Cherries are good sources of bioactive phenolic compounds that are widely considered to be potentially healthy. Here we investigated the protective activities of juice and wine products of tart and sweet cherries and their constituent anthocyanins (e.g., cyanidin 3-glucoside and cyanidin 3-rutinoside) against oxidative stress induced by hydrogen peroxide (H₂O₂) in Chinese hamster lung fibroblasts (V79-4). Total phenolics in the cherry juices and wines were 56.7–86.8 mg of gallic acid equivalents (GAE)/l and 79.4–149 mg GAE/l, respectively. Total anthocyanins in the cherry juices and wines were 7.9–50.1 mg of cyanidin 3-glucoside equivalents (CGE)/l and 29.6–63.4 mg CGE/l, respectively. Both cherry juices and wines exerted protective effects against oxidative stress induced by H₂O₂ on V79-4 cells and also enhanced the activities of antioxidative enzymes, such as superoxide dismutase and catalase, in a dose-dependent manner. The protection of V79-4 cells from oxidative stress by phenolics was mainly attributable to anthocyanins. The positive correlation between the protective effects against oxidative stress in V79-4 cells and the antioxidant enzyme activities was stronger for cyanidin 3-glucoside than for cyanidin 3-rutinoside.     

Oxidative stress prevention and anti-apoptosis activity of grape (Vitis vinifera L.) stems in human keratinocytes

To date, grape stems have been partially assessed on their content in phenolics and their radical scavenging activity, whilst the potential to modulate oxidative stress in biological models remains underexplored. In the present work, the effect of grape stems' phenolics on redox unbalance was evaluated in human keratinocytes (HaCaT cells). Grape stems' extracts were assessed on their phenolic composition by high performance liquid chromatography coupled with photodiode array detection and electrospray ionization-mass spectrometry (HPLC–PAD-ESi-MSn), besides on radical scavenging capacity (ABTS and DPPH). In addition, their protective effect against oxidative stress induced by H₂O₂ by the determination of the level of glutathione, reactive oxygen species, lipid peroxidation, and overall oxidative stress in HaCaT cells by flow cytometry was evaluated. This characterization allowed to identify five flavonols, one cinnamic acid, and one stilbene. A close correlation between the concentration of these phenolics and the capacity to scavenge free radicals and with the potential to modulate the redox balance in vitro was observed. From the analysis of correlation, the activity of malvidin-3-O-glucoside, malvidin-3-O-(6-O-caffeoyl)-glucoside, and malvidin-3-O-rutinoside with respect to the prevention of basal oxidative stress and the capacity of isorhamnetin-3-O-(6-O-feruloyl)-glucoside and kaempferol-3-O-rutinoside to prevent H₂O₂-induced redox unbalance were stated. Furthermore, grape stems' phenolics also showed an efficient capacity to modulate apoptosis in HaCaT cells, reducing the frequency of annexin V/PI double positive apoptotic cells by up to 99.5% relative to controls, which was further confirmed by the determination of the appearance of the occurrence of apoptotic bodies and the expression of activated (cleaved) caspase-3 by flow cytometry and western-blot, respectively. These results supported the potential of individual phenolics from grape stems to modulate oxidative stress, allowing to envisage dedicated combinations of single compounds for the development of efficient formulations efficient against oxidative stress.     

Protective effect of whey protein hydrolysates on H2O2-induced PC12 cells oxidative stress via a mitochondria-mediated pathway

Whey protein hydrolysates (WPHs) were prepared with pepsin and trypsin. A PC12 cell model was built to observe the protective effect of WPHs against H₂O₂-induced oxidative stress. The results indicated that WPHs reduced apoptosis by 14% and increased antioxidant enzyme activities. Flow cytometry was used to assess the accumulation of reactive oxygen species (ROS), Ca²⁺ levels and the mitochondrial membrane potential (MMP). The results showed that WPHs suppressed ROS elevation and Ca²⁺ levels and stabilised MMP by 16%. The anti-apoptosis/pro-apoptosis proteins Bcl-2/Bax and poly (ADP-ribose) polymerase (PARP) were investigated by Western-blot analysis, which indicated that WPHs increased the expression of Bcl-2 while inhibiting the expression of Bax and the degradation of PARP. WPHs also blocked Caspase-3 activation by 62%. The results demonstrate that WPHs can significantly protect PC12 cells against oxidative stress via a mitochondria-mediated pathway. These findings indicate the potential benefits of WPHs as valuable food antioxidative additives.     

Curcumin protects mouse neuroblastoma Neuro-2A cells against hydrogen-peroxide-induced oxidative stress

Curcumin has been traditionally used in China and India for food and medicinal purposes. It has been shown to possess potent antioxidative activity both in vitro and in vivo. In the present study, the neuroprotective effects and the potential mechanisms of curcumin against H₂O₂-induced oxidative stress in mouse neuroblastoma Neuro-2A cells were investigated. Treatment with curcumin at 20 and 25 μg/mL for 1 h prior to H₂O₂ exposure significantly attenuated cell viability loss, reduced apoptosis, suppressed the elevation of intracellular reactive oxygen species (ROS) and calcium levels, and stabilised mitochondrial membrane potential. Furthermore, curcumin could block H₂O₂-mediated degradation of the protein IκBα and subsequent activation of nuclear factor κB, thus inhibiting the expression of its target gene cyclooxygenase 2. These results indicate that curcumin has potential protective effects against H₂O₂-induced oxidative stress in neuron cells, which might make curcumin a suitable therapeutic agent for prevention and treatment of neurodegenerative diseases associated with oxidative stress.     

Induction of apoptosis by Myagropsis myagroides extracts is associated with a caspase dependent pathway and reactive oxygen species generation in human cancer cells

The purpose of this study was to elucidate the mechanisms underlying apoptosis induced by an ethanol extracts from Myagropsis myagroides (ME) in HeLa, U937, and PC-3 cells. ME treatment for 24 h significantly inhibited cell viability in a dose-dependent manner, and induced apoptosis. Moreover, ME treatment triggered the cleavage of caspase-8, ?9, ?3, and poly(ADP-ribose) polymerase (PARP). A general caspase inhibitor (z-VAD-fmk) inhibited ME-induced activation of caspase-3, PARP cleavage, and cell death. ME treatment also triggered the release of cytochrome c from the mitochondria to the cytosol and stimulated the cleavage of Bid, up-regulation of Bax, and down-regulation of Bcl-2. Furthermore, ME treatment caused reactive oxygen species (ROS) generation. An antioxidant N-acetylcysteine (NAC) blocked MEinduced activation of caspase-3, PARP cleavage, and cell death. Overall, these results suggest that ME-induced apoptosis is mediated by a caspase dependent pathway and ROS generation in HeLa, U937, and PC-3 cells.     

Selenomethionine increases proliferation and reduces apoptosis in bovine mammary epithelial cells under oxidative stress

The decline in mammary epithelial cell number as lactation progresses may be due, in part, to oxidative stress. Selenium is an integral component of several antioxidant enzymes. The present study was conducted to examine the effect of oxidative stress and selenomethionine (SeMet) on morphology, viability, apoptosis, and proliferation of bovine mammary epithelial cells (BMEC) in primary culture. Cells were isolated from mammary glands of lactating dairy cows and grown for 3 d in a low-serum gel system containing lactogenic hormones and 0 or 100 μM H₂O₂ with 0, 10, 20, or 50 nM SeMet. Hydrogen peroxide stress increased intracellular H₂O₂ to 3 times control concentrations and induced a loss of cuboidal morphology, cell-cell contact, and viability of BMEC by 25%. Apoptotic cell number more than doubled during oxidative stress, but proliferating cell number was not affected. Supplementation with SeMet increased glutathione peroxidase activity 2-fold and restored intracellular H₂O₂ to control levels with a concomitant return of morphology and viability to normal. Apoptotic BMEC number was decreased 76% below control levels by SeMet and proliferating cell number was increased 4.2-fold. These findings suggest that SeMet modulated apoptosis and proliferation independently of a selenoprotein-mediated reduction of H₂O₂. In conclusion, SeMet supplementation protects BMEC from H₂O₂-induced apoptosis and increased proliferation and cell viability under conditions of oxidative stress.     

Analysis of phenolic compounds and antioxidant activity with H4IIE cells of three different rice grain varieties

The phenolic content and the antioxidant capacity of ethanolic extract of rice grains were examined using three different rice (Oryza sativa L. var. japonica) varieties, Jakwangchalbyeo (red pericarp glutinous rice), Hwasunchalbyeo (white pericarp glutinous rice), and Ilpumbyeo (white pericarp non-glutinous rice). Quantitative high performance liquid chromatography (HPLC) analyses showed that Jakwangchalbyeo contains the highest amount of total phenolic substances. Radical scavenging activities measured by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) assay were 87.5% (Jakwangchalbyeo), 45.0% (Hwasunchalbyeo), and 50.0% (Ilpumbyeo). The results suggest a positive correlation between the phenolic content and the antioxidation capacity. Antioxidant activity in the ethanolic extract of rice grain was further assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assay and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H₂O₂). Hwasunchalbyeo and Ilpumbyeo extracts showed no significant effects on H₂O₂-induced oxidative stress. Jakwangchalbyeo extract increased cell viability by up to 82% and 74% after 5- and 24-h treatments at 100 μg/mL, respectively. Consistent with the results of DPPH and MTT assays, dichlorofluorescein (DCF) fluorescence was further reduced when H₂O₂ was applied with Jakwangchalbyeo extract. In conclusion, red pericarp Jakwangchalbyeo grain extracts exerted significantly higher inhibitory effects on H₂O₂-induced oxidative stress in H4IIE cells than did other white pericarp rice grain extracts.     

Cytoprotective effect of white tea against H2O2-induced oxidative stress in vitro

The protective effect of water extracts of white tea (WEWT) on oxidative stress in vitro is investigated. WEWT, like water extracts of green tea (WEGT) and water extracts of Pu-erh tea (WEPT), demonstrates a marked inhibition of the oxidation of liposome, albumin and LDLmodel systems. WEWT protects against H₂O₂-induced cytotoxicity, in a dose-dependent manner. The inhibition of ROS generation and MDA formation by WEWT in H₂O₂-induced Clone 9 cells parallels the effects on cell viability. Moreover, GSH and antioxidant enzymes may play an important role in the protective effect that is associated with H₂O₂-induced oxidative stress. The HPLC-DAD and HPLC–MS/MS analysis, shows that sixteen bioactive compounds are present in WEWT, which may partially account for its protective effect against oxidative insult. These results suggest that the mechanism of the protective actions of WEWT is related to its antioxidant potential and the maintenance of the normal redox status of the cell.     

Effects of hydrogen peroxide and l-tryptophan on antioxidative potential,apoptosis, and mammalian target of rapamycin signaling in bovine intestinal epithelial cells

Amino acids are primarily absorbed in the ruminant small intestine, and the small intestine is a target organ prone to oxidative stress, causing intestinal disfunction. Previous study suggested that l-Trp could benefit intestinal function and production performance. This study aimed to explore the effects of l-Trp on hydrogen peroxide (H₂O₂)-induced oxidative injury in bovine intestinal epithelial cells (BIEC) and the potential mechanism. The effects of l-Trp on cell apoptosis, antioxidative capacity, AA transporters, and the mammalian target of rapamycin (mTOR) signaling pathway were evaluated in BIEC treated with 0.8 mM l-Trp for 2 hours combined with or without H₂O₂ induction. In addition, to explore whether the effects of 0.8 mM l-Trp on oxidative stress were related to mTOR, an mTOR-specific inhibitor was used. The percentage of apoptosis was measured using flow cytometry. The relative gene abundance and protein expression in BIEC were determined using real-time PCR and Western blot assay, respectively. Results showed l-Trp at 0.4 and 0.8 mM enhanced the cell viability, and it was inhibited by l-Trp at 6.4 mM. l-Tryptophan at 0.4, 0.8, and 1.6 mM remarkably decreased the percentage of apoptosis and enhanced antioxidative capacity in H₂O₂-mediated BIEC. Moreover, l-Trp at 0.8 mM increased the relative gene abundance and protein expression of antioxidative enzymes and AA transporters, and the mTOR signaling pathway. The mTOR inhibitor lowered the protein expression of large neutral amino acid transporter 1, but the inhibition of mTOR did not alter the activities of catalase and superoxide dismutase or protein expression of alanine-serine-cysteine transporter 2 with or without H₂O₂ induction. l-Tryptophan increased catalase and superoxide dismutase activities in H₂O₂-mediated BIEC, although not with a present mTOR inhibitor. l-Tryptophan increased the protein expression of large neutral amino acid transporter 1 and alanine-serine-cysteine transporter 2 in H₂O₂-mediated BIEC with or without the presence of an mTOR inhibitor. The present work suggested that l-Trp supplementation could alleviate oxidative injury in BIEC by promoting antioxidative capacity and inhibiting apoptosis, and the mTOR signal played vital roles in the alleviation.     

Regulation of apoptosis in the atresia of dominant bovine follicles of the first follicular wave following ovulation

During atresia of bovine follicles, granulosa cells are lost through the controlled form of cell death, apoptosis. The purpose of this study was to characterize the regulation of apoptotic death of granulosa cells in dominant bovine follicles during the first wave of follicular development. Dominant follicles were collected from Holstein heifers on days 4, 6 or 8 of the first follicular wave (n = 5/day). Regulation of apoptosis in granulosa cells was examined by annexin V and propidium iodide staining; measurement of relative levels of mRNA encoding Bcl-2, Bcl-xL and Bax; and activity of caspase-3, -8 and -9. Steady-state levels of mRNA encoding four oxidative stress-response proteins were determined. Compared with day 4, the incidence of apoptotic and nonviable granulosa cells tended to increase on day 6, and numbers of nonviable cells were higher on day 8. The ratios of relative levels of mRNA encoding Bcl-2 to Bax and Bcl-xL to Bax were higher on day 6 than days 4 and 8. Activity of caspases-3 and -9 in granulosa cells did not change among the 3 days, while caspase-8 activity decreased on day 8 compared with days 4 and 6. Amounts of GSHPx, MnSOD and Cu/ZnSOD mRNA in granulosa cells were higher on day 8 than day 6. In theca interna, amounts of Cu/ZnSOD mRNA decreased between days 4 and 6. From the decreased production of estradiol and increased numbers of apoptotic and nonviable granulosa cells, we conclude that atresia of the dominant follicle is initiated between days 4 and 6 of the first follicular wave. However, apoptosis of granulosa cells does not appear to be initiated by changes in expression of oxidative stress-response proteins.     

Catechin-7-O-β-d-glucopyranoside scavenges free radicals and protects human B lymphoma BJAB cells on H2O2-mediated oxidative stress

Catechin-7-O-β-d-glucopyranoside (CA-G) was previously isolated from red bean (the seed of Phaseolus calcaratus cv. Roxburgh). This study examined the ability of CA-G to scavenge reactive oxygen species generated by cell-free systems and to protect cells from oxidative stress caused by hydrogen peroxide (H₂O₂). The mechanism by which CA-G exerts its antioxidant and anti-apoptotic action on H₂O₂-exposed cells was also investigated. CA-G treatment prevented H₂O₂-mediated apoptosis and inhibited the formation of single stand breaks in DNA in H₂O₂-exposed BJAB cells. CA-G suppressed mitochondrial stress and caspase activation caused by H₂O₂. Mechanistic experiments revealed that the antioxidant mechanism of CA-G on H₂O₂-mediated oxidative damage was due to the direct scavenging of hydroxyl radicals and/or to the chelation of metal ions that were used to produce hydroxyl radicals from H₂O₂ via the Fenton reaction. Collectively, these findings suggest beneficial roles of CA-G or CA-G-rich red bean on the protection from oxidative damage.     

Oryzadine,a new alkaloid of Oryza sativa cv. Heugjinjubyeo, attenuates oxidative stress-induced cell damage via a radical scavenging effect

The antioxidant properties of oryzadine, a new alkaloid, obtained from Oryza sativa cv. Heugjinjubyeo was investigated by applying various methods based on cell-free and cell experiments. Oryzadine showed scavenging effects on the hydroxyl radical, superoxide radical, and intracellular reactive oxygen species (ROS). Oryzadine inhibited H₂O₂-induced DNA damage, which was demonstrated by DNA tail formation, lipid peroxidation which was demonstrated by the formation of thiobarbituric acid reactive substance (TBARS), and protein oxidation which was demonstrated by protein carbonyl formation. Therefore, oryzadine protected H₂O₂-induced cell damage. Our results show that the cytoprotective effects of oryzadine stem from its ability to inhibit H₂O₂-induced apoptosis, as demonstrated by a decrease in apoptotic body formation and the inhibition of mitochondrial membrane potential (ΔΨ) loss. Taken together, these findings suggest that oryzadine protected cells against H₂O₂-induced cell damage via ROS scavenging effect. Therefore, oryzadine could be considered a significant natural source of antioxidant.     

Comparison between conjugated linoleic acid and essential fatty acids in preventing oxidative stress in bovine mammary epithelial cells

Some in vitro and in vivo studies have demonstrated protective effects of conjugated linoleic acid (CLA) isomers against oxidative stress and lipid peroxidation. However, only a few and conflicting studies have been conducted showing the antioxidant potential of essential fatty acids. The objectives of the study were to compare the effects of CLA to other essential fatty acids on the thiol redox status of bovine mammary epithelia cells (BME-UV1) and their protective role against oxidative damage on the mammary gland by an in vitro study. The BME-UV1 cells were treated with complete medium containing 50 μM of cis-9,trans-11 CLA, trans-10,cis-12 CLA, α-linolenic acid, γ-linolenic acid, and linoleic acid. To assess the cellular antioxidant response, glutathione, NADPH, and γ-glutamyl-cysteine ligase activity were measured 48 h after addition of fatty acids (FA). Intracellular reactive oxygen species and malondialdehyde production were also assessed in cells supplemented with FA. Reactive oxygen species production after 3 h of H₂O₂ exposure was assessed to evaluate and to compare the potential protection of different FA against H₂O₂-induced oxidative stress. All FA treatments induced an intracellular GSH increase, matched by high concentrations of NADPH and an increase of γ-glutamyl-cysteine ligase activity. Cells supplemented with FA showed a reduction in intracellular malondialdehyde levels. In particular, CLA isomers and linoleic acid supplementation showed a better antioxidant cellular response against oxidative damage induced by H₂O₂ compared with other FA.     

Evidence for protective effects of coffees on oxidative stressinduced apoptosis through antioxidant capacity of phenolics

This study evaluated total phenolics, total flavonoids, and antioxidant capacity of randomly selected regular and decaffeinated coffees commercially available in Korea and their protective effects in human hepatic epithelial HepG2 cell line against oxidative stress. All coffees tested exhibited potent antioxidant capacity in chemical systems and, consequently, significant protection of cells from oxidative stress in vitro in a dose-dependent manner. In particular, H₂O₂-induced apoptosis as evaluated by annexin V staining and flow cytometry was prevented by coffee extracts, resulting in the enhanced cell viability. Of interest, the content of total phenolics and flavonoids in coffees demonstrated a positive correlation with antioxidant capacity, indicating that the antioxidant capacity of coffees may be attributed to those phytochemicals. In accordance with previous studies, caffeoylquinic acid (CQA) and its derivatives including 3-CQA, 4-CQA, 5-CQA, 3,4-diCQA, 3,5-diCQA, and 4,5-diCQA were identified as phenolic phytochemicals by a reversed-phase HPLC, with 5-CQA being a major component. Taken together, the present study demonstrated protective effects of regular and decaffeinated coffees on cells in vitro against overwhelming oxidative stress due to richness in phenolics, especially CQA and its derivatives. Coffees, regular or decaffeinated, may serve as a good source of health-beneficial phytochemicals in diet.     

Cytoprotective activity of extract of roasted coffee residue on mouse embryonic fibroblasts cells against apoptosis induced by oxidative stress

This study was conducted to evaluate the cytoprotective activity of roasted coffee residues (RCRs) extract on mouse embryonic fibroblast (MEF) cells. RCRs originated from Colombia and Honduras are relatively nontoxic to cell growth and even stimulate cell proliferation. Colombian RCRs showed most efficient protective effects on MEF cells against oxidative damage induced by H₂O₂ compared among the extracts prepared under the same roasting time. The most significant radical scavenging activity was measured in RCR with roasting time of 8.5 min. Phenolic and nonphenolic compounds in RCRs were chlorogenic acid, caffeine, caffeic acid, nicotinic acid, trigonelline, and 5-(hydroxymethyl)furfuralolehyde. Effect of Colombian RCRs on apoptosis occurred by oxidative damage was evaluated by morphological and flow cytometric analysis. Apoptotic cell accumulation was decreased by cotreatment of MEF cells with Colombian RCRs. These results suggested that antioxidant potency of RCRs suppresses the cytotoxicity which is induced by H₂O₂ and has a protective effect on MEF cell against oxidative stress.     

Free radical scavenging activity and comparative proteomic analysis of antioxidative protein against H2O2-induced oxidative stress in neuronal cells

Artemisia annua was enzymatically hydrolyzed by five proteases and seven carbohydrases. All enzymatic extracts scavenged DPPH, hydroxyl and alkyl radicals. Especially, the Protamex among the various proteases and Maltogenase among the various carbohydrases extracts exhibited the highest scavenging activity on hydroxyl radical. The extracts of A. annua clearly reduced neuronal cell death from H₂O₂-induced damage. In addition, a proteomic analysis, two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionisation-time of flight/time of flight (MALDI-TOF/TOF) was used to identify the proteins of the neuronal cells whose expressions were or were not altered by the treatment of the Maltogenase extracts which showed the highest hydroxyl radical scavenging activity among all enzymatic extracts for 24 h. The protein characterisation revealed that translation elongation factor Tu (EF-Tu), Immunoglobulin E (IgE) and voltage-dependent anion channel 1 (VDAC-1) were involved in the cell survival effects against H₂O₂-induced apoptosis. These results suggest that EF-Tu, IgE and VDAC-1 have an important role in the reduction of neuronal apoptosis by oxidative stress, and the enzymatic extracts of A. annua shows potent antioxidative activities by regulating EF-Tu, IgE and VDAC-1.