H2O2-induced Oxidative Stress Damage Research Articles

Protective Effect of Arzanol against H2O2-Induced Oxidative Stress Damage in Differentiated and Undifferentiated SH-SY5Y Cells.

Oxidative stress can damage neuronal cells, greatly contributing to neurodegenerative diseases (NDs). In this study, the protective activity of arzanol, a natural prenylated α-pyrone-phloroglucinol heterodimer, was evaluated against the H2O2-induced oxidative damage in trans-retinoic acid-differentiated (neuron-like) human SH-SY5Y cells, widely used as a neuronal cell model of neurological disorders. The pre-incubation (for 2 and 24 h) with arzanol (5, 10, and 25 μM) significantly preserved differentiated SH-SY5Y cells from cytotoxicity (MTT assay) and morphological changes induced by 0.25 and 0.5 mM H2O2. Arzanol reduced the generation of reactive oxygen species (ROS) induced by 2 h oxidation with H2O2 0.5 mM, established by 2',7'-dichlorodihydrofluorescein diacetate assay. The 2 h incubation of differentiated SH-SY5Y cells with H2O2 determined a significant increase in the number of apoptotic cells versus control cells, evaluated by propidium iodide fluorescence assay (red fluorescence) and NucView® 488 assay (green fluorescence). Arzanol pre-treatment (2 h) exerted a noteworthy significant protective effect against apoptosis. In addition, arzanol was tested, for comparison, in undifferentiated SH-SY5Y cells for cytotoxicity and its ability to protect against H2O2-induced oxidative stress. Furthermore, the PubChem database and freely accessible web tools SwissADME and pkCSM-pharmacokinetics were used to assess the physicochemical and pharmacokinetic properties of arzanol. Our results qualify arzanol as an antioxidant agent with potential neuroprotective effects against neuronal oxidative stress implicated in NDs.

Quercetin Alleviates H2O2-Induced Oxidative Stress Damage to Human Endometrial Stromal Cells by Inhibiting the p38 MAPK/NOX4 Signaling Pathway

Quercetin Alleviates H2O2-Induced Oxidative Stress Damage to Human Endometrial Stromal Cells by Inhibiting the p38 MAPK/NOX4 Signaling Pathway

Cytisine eye drops for benzalkonium chloride-induced dry eye: safety and efficacy evaluation

This experiment aimed to investigate the feasibility of cytisine (CYT) in treating eye diseases with ocular topical application. An in vitro cytotoxicity test, a hen’s egg test-chorioallantoic membrane (HET-CAM), and a mouse eye tolerance test were used to fully reveal the ocular safety profiles of CYT. For the efficacy evaluations, CYT’s effects on cell wound healing, against H2O2-induced oxidative stress damages on cells, and on benzalkonium chloride (BAC)–induced dry eye disease (DED) in mice were evaluated. Results showed that CYT did not show any cytotoxicities at concentrations no higher than 250 μg/ml, while lipoic acid (α-LA) at 250 μg/ml and BAC at 1.25 μg/ml showed significant cytotoxicities within 48 h incubation. The HET-CAM and mouse eye tolerance test confirmed that 0.5% CYT eye drops demonstrated good safety characteristics. Efficacy evaluations showed that CTY significantly promoted cell migration and wound healing. CYT significantly improved cell survival against H2O2-induced oxidative stress damage by reversing the imbalance between the reactive oxygen species (ROS) and antioxidant defense mechanisms. The animal evaluation of the BAC-induced dry eye model revealed that CYT demonstrated a strong treatment effect, including reversing ocular surface damages, recovering corneal sensitivity, and inhibiting neovascularization; HMGB1/NF-κB signaling was involved in this DED treatment by CTY. In conclusion, CYT had strong experimental treatment efficacy against DED with good ocular safety profiles, and it might be a novel and promising drug for DED.

Study on the Protective Effect of Flavonoids in Vernonia anthelmintica (L.) Willd on Oxidative Stress Damage of HaCaT Cells

Background: Vitiligo, a prevalent hypopigmented skin disorder, profoundly impacts patients’ psychological well-being and quality of life. Therefore, the development of more efficacious antivitiligo drugs is imperative. Objectives: The aim of this study was to investigate the protective effects of four flavonoids found in Vernonia anthelmintica (L.) Willd such as Butin, Butein, Eriodictyol, and Liquiritigenin—against H2O2-induced oxidative stress damage in human immortalized epidermal cells (HaCaT cells). Materials & Methods: Cytotoxicity was assessed using the MTT kit. Superoxide dismutase (SOD) activity was measured via the WST-1 method, while the content of malondialdehyde (MDA) was determined using the thiobarbituric acid method. Transcription and protein expression of nuclear transcription-related factors (Nrf2), heme oxidase-1 (HO-1), and Quinone oxidoreductase (NQO1) were analyzed through quantitative real-time polymerase chain reaction and Western Blotting. Results: Upon treatment with the four flavonoids, SOD activity exhibited a significant increase compared to the H2O2 induction model ( P < .01), and MDA content was significantly lower ( P < .01). This effect showed a dose-dependent relationship, with an increment in SOD activity and a reduction in MDA content. All four flavonoids upregulated the expression levels of nuclear transcription-related factor, HO-1, and NQO1 genes, as well as their corresponding proteins. Conclusion: The identified flavonoids in V anthelmintica (L.) Willd demonstrate a protective effect against H2O2-induced oxidative stress damage in HaCaT cells.

Collagen Scaffolds Functionalized by Cu2+-Chelated EGCG Nanoparticles with Anti-Inflammatory, Anti-Oxidation, Vascularization, and Anti-Bacterial Activities for Accelerating Wound Healing.

Skin injury is a common health problem worldwide, and the highly complex healing process poses critical challenges for its management. Therefore, wound dressings with salutary effects are urgently needed for wound care. However, traditional wound dressing with a single function often fails to meet the needs of wound repair, and the integration of multiple functions has been required for wound repair. Herein, Cu2+-chelated epigallocatechin gallate nanoparticles (EAC NPs), with radical scavenging, inflammation relieving, bacteria restraining, and vascularization accelerating capacities, are adopted to functionalize collagen scaffold, aiming to promote wound healing. Radical scavenging experiments verify that EAC NPs could efficiently scavenge radicals. Additionally, EAC NPs could effectively remove Escherichia coli and Staphylococcus aureus. H2O2 stimuli-responsive EAC NPs show slow and sustained release properties of Cu2+. Furthermore, EAC NPs exhibit protective effects against H2O2-induced oxidative-stress damage and anti-inflammatory activity in vivo. Physicochemical characterizations show that the introduction of EAC NPs does not disrupt the gelation behavior of collagen, and the composite scaffolds (CS) remain porous structure similar to collagen scaffold. Animal experiments demonstrate that CS could promote wound healing through improving the thickness of renascent epidermis and number of new vessels. CS with multiple salutary functions is a promising dressing for wound care.

Preparation and identification of novel antioxidant peptides from camel bone protein.

Collagen hydrolysates are a vital source of bioactive peptides. The objective of this study was to prepare camel bone collagen hydrolysates with antioxidant activity, and to identify the peptides responsible for the antioxidant activity. To this end, single-factor and orthogonal tests were performed to explore the optimum preparation conditions. A hydrolysis time of 5h, enzyme:substrate ratio of 1200 U/g, pH of 7.0, and a material:water ratio of 1:3.0 were adopted. Subsequently, the hydrolysates were purified using a series of chromatography procedures, and three novel peptides, GPPGPPGPPGPPGPPSGGFDF (hydroxylation), PATGDLTDFLK, and GSPGPQGPPGSIGPQ, possessing antioxidant abilities, were identified from the fraction using liquid chromatography-tandem mass spectrometry. The peptide PATGDLTDFLK showed excellent DPPH scavenging activity (39%) and a good cytoprotective effect on H2O2-induced oxidative stress damage in HepG2 cells with a 21.1% increase observed.

Protective Effect of Apple Polyphenols on H2O2-Induced Oxidative Stress Damage in Human Colon Adenocarcinoma Caco-2 Cells.

Apple is an important dietary agent for human and apple polyphenols (AP) are the main secondary metabolites of apples. In this study, the protective effects of AP on hydrogen peroxide (H2O2)-induced oxidative stress damage in human colon adenocarcinoma Caco-2 cells were investigated by cell viability, oxidative stress change as well as cell apoptosis. Pre-adding AP could significantly increase the survival rate of H2O2-treated Caco-2 cells. Besides, the activities of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) were elevated. While the malondialdehyde (MDA) content which is the major oxidant products of polyunsaturated fatty acids (PUFA) reduced after AP treatment. In addition, AP also suppressed the emergence of DNA fragment and decreased the expression of apoptosis-related protein Caspase-3. These results demonstrated that AP could ameliorate H2O2-induced oxidative stress damage in Caco-2 cells, which could serve as a reference for further studies of apple natural active products and deep study of the anti-oxidative stress mechanism.

Chemical composition of the Kaempferia galanga L. essential oil and its in vitro and in vivo antioxidant activities.

Oxidative stress is closely related to the development of many diseases. Essential oils (EOs) show potent antioxidant activity from natural sources. Kaempferia galanga L. is an important medicine rich in high-value essential oil (KGEO). However, the antioxidant activity of KGEO remains to be fully studied. Chemical composition of KGEO was analyzed using gas chromatography-mass spectrometry (GC-MS). The antioxidant activity was determined using the DPPH, ABTS, hydroxyl radical scavenging assays and reducing power assay in vitro. A zebrafish model was used to evaluate the protective effect of KGEO against H2O2-induced oxidative stress damage in vivo. The major components of KGEO were found to be trans ethyl p-methoxycinnamate (32.01%), n-pentadecane (29.14%) and trans ethyl cinnamate (19.50%). In vitro pharmacological results showed that KGEO had good free radical scavenging capacity in DPPH, ABTS, and hydroxyl radical scavenging assays (IC50 values: 19.77 ± 1.28, 1.41 ± 0.01, and 3.09 ± 0.34 mg/mL, respectively) and weak reducing capacity in the reducing power assay (EC50 value: 389.38 ± 4.07 mg/mL). In vivo zebrafish experiments results indicated that the survival rate and heart rate increased, and ROS generation, cell death, and lipid peroxidation were attenuated after KGEO treatment. In addition, a decrease in malondialdehyde (MDA) levels and increases in superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were observed in the KGEO-treated groups. This study validated the in vitro and in vivo antioxidant activities of KGEO, which provides a theoretical basis for a profound study of KGEO and its application in the pharmaceutical, food and cosmetic industries.

NMR-Based Metabolomic Analysis for the Effects of Trimethylamine N-Oxide Treatment on C2C12 Myoblasts under Oxidative Stress.

The gut microbial metabolite trimethylamine N-oxide (TMAO) has received increased attention due to its close relationship with cardiovascular disease and type 2 diabetes. In previous studies, TMAO has shown both harmful and beneficial effects on various tissues, but the underlying molecular mechanisms remain to be clarified. Here, we explored the effects of TMAO treatment on H2O2-impaired C2C12 myoblasts, analyzed metabolic changes and identified significantly altered metabolic pathways through nuclear magnetic resonance-based (NMR-based) metabolomic profiling. The results exhibit that TMAO treatment partly alleviated the H2O2-induced oxidative stress damage of cells and protected C2C12 myoblasts by improving cell viability, increasing cellular total superoxide dismutase capacity, improving the protein expression of catalase, and reducing the level of malondialdehyde. We further showed that H2O2 treatment decreased levels of branched-chain amino acids (isoleucine, leucine and valine) and several amino acids including alanine, glycine, threonine, phenylalanine and histidine, and increased the level of phosphocholine related to cell membrane structure, while the TMAO treatment partially reversed the changing trends of these metabolite levels by improving the integrity of the cell membranes. This study indicates that the TMAO treatment may be a promising strategy to alleviate oxidative stress damage in skeletal muscle.

Hemicellulosic Polysaccharides From Bamboo Leaves Promoted by Phosphotungstic Acids and Its Attenuation of Oxidative Stress in HepG2 Cells.

In this work, we exploited an efficient method to release hemicellulosic polysaccharides (BLHP) from bamboo (Phyllostachys pubescens Mazel) leaves assisted by a small amount of phosphotungstic acid. Structural unit analysis proved that BLHP-A1 and BLHP-B1 samples possessed abundant low-branch chains in →4)-β-D-Xylp-(1→ skeleton mainly consisting of Xylp, Manp, Glcp, Galp, and Araf residues. According to the results of the antioxidant activity assays in vitro, both of the two fractions demonstrated the activity for scavenging DPPH⋅ and ABTS+ radicals and exhibited relatively a high reducing ability compared to the recently reported polysaccharides. Moreover, the antioxidant activities of purified polysaccharides were evaluated against H2O2-induced oxidative stress damage in HepG2 cells. BLHP-B1 showed more activity for preventing damages from H2O2 in HepG2 cells by improving the enzyme activities of SOD, CAT, and GSH-Px and decreasing the production of MDA as well as suppressing reactive oxygen species (ROS) formation. This study implied that BLHP could demonstrate its attenuation ability for oxidative stress in HepG2 cells.

Melatonin Alleviates Oxidative Stress Induced by H2O2 in Porcine Trophectoderm Cells.

Placental oxidative stress has been implicated as a main risk factor for placental dysfunction. Alleviation of oxidative stress and enhancement of antioxidant capacity of porcine trophectoderm (PTr2) cells are effective means to maintaining normal placental function. The present study was conducted to evaluate the protective effect of melatonin (MT) on H2O2-induced oxidative damage in PTr2 cells. Our data revealed that pretreatment with MT could significantly improve the decrease in cell viability induced by H2O2, and reduce intracellular reactive oxygen species (ROS) levels and the ratio of apoptotic cells. Here, we compared the transcriptomes of untreated versus melatonin-treated PTr2 cells by RNA-seq analysis and found that differentially expressed genes (DEGs) were highly enriched in the Wnt signaling, TGF-beta signaling and mTOR signaling pathways. Moreover, pretreatment with MT upregulated the antioxidant-related genes such as early growth response3 (EGR3), WAP four-disulfide core domain1 (WFDC1), heme oxygenase1 (HMOX1) and vimentin (VIM). These findings reveal that melatonin protects PTr2 cells from H2O2-induced oxidative stress damage.

Quercetin Improves Mitochondrial Function and Inflammation in H2O2-Induced Oxidative Stress Damage in the Gastric Mucosal Epithelial Cell by Regulating the PI3K/AKT Signaling Pathway.

Functional dyspepsia (FD) is one of the most common functional gastrointestinal disorders, the therapeutic strategy of which it is limited due to its complex pathogenesis. Oxidative stress-induced damage in gastric mucosal epithelial cells is related to the pathogenesis and development of FD. Quercetin (Que) is one of the active ingredients of Zhishi that showed antioxidant, antiapoptotic, and anti-inflammatory effects. The aim of this study is to investigate the effect of Que on oxidative stress-induced gastric mucosal epithelial cells damage and its underlying molecular mechanism. The gastric mucosal epithelial cell line GES-1 was treated with 200 μM of H2O2 to construct an oxidative stress-induced damage model. The H2O2 cells were then administrated with different concentrations of Que. The results indicated that high concentration of Que (100 μM) showed cytotoxicity in H2O2-induced GES-1 cells. However, appropriate concentration of Que (25 and 50 μM) alleviated the oxidative stress damage induced by H2O2, as demonstrated by the increase of proliferation, decrease of ROS generation, apoptosis, inflammation, and alleviation of mitochondrial function and cell barrier. In addition, Que increased the activation of phosphorylation of PI3K and AKT decreased by H2O2. To investigate whether Que alleviated the oxidative stress damage in GES-1 cells by the PI3K/AKT signaling pathway, the GES-1 cells were treated with Que (25 μM) combined with and without LY294002, the PI3K inhibitor. The results showed that LY294002 suppressed the alleviation effect on Que in H2O2-induced GES-1 cells. In conclusion, the current study demonstrates that Que alleviates oxidative stress damage in GES-1 cells by improving mitochondrial function and mucosal barrier and suppressing inflammation through regulating the PI3K/AKT signaling pathway, indicating the potential therapeutic effects of Que on FD.

Chemical compositions, anti-oxidant and anti-inflammatory potential of ethanol extract from Zhuke-Hulu tea

Zhuke-Hulu tea is an herbal tea widely enjoyed by people in Southern China. For the first time the phytochemical compositions of Zhuke-Hulu tea ethanol extract (ZHTE) were analyzed by GC-MS and UPLC-ESI-MS/MS, and the anti-oxidant and anti-inflammatory activities were evaluated. Several categories of substances in ZHTE were identified, among which organic acids, phenolics/phenolic acids and flavonoids were the major components. ZHTE showed significant anti-oxidant activity as measured by free radical scavenging assays and reducing power test. Moreover, ZHTE exhibited cytoprotective effect against H2O2-induced oxidative stress damage by enhancing endogenous anti-oxidant defense system and Nrf2 nuclear translocation. In addition, ZHTE also exerted anti-inflammatory efficiency by down-regulating the protein levels of iNOS and COX-2 as well as the production of pro-inflammatory cytokines. In parallel, ZHTE alleviated intestinal inflammation induced by dextran sulfate sodium (DSS) through reducing pro-inflammatory cytokines level, inhibiting inflammatory cell infiltration, and modulating gut microbiota. These findings provide a solid foundation for the nutraceutical benefits of Zhuke-Hulu tea and promote its consumption as a functional beverage.

Design, green synthesis, antioxidant activity screening, and evaluation of protective effect on cerebral ischemia reperfusion injury of novel monoenone monocarbonyl curcumin analogs

Antioxidants with high efficacy and low toxicity have the potential to treat cerebral ischemia reperfusion injury (CIRI). Dienone monocarbonyl curcumin analogs (DMCA) capable of overcoming the instability and pharmacokinetic defects of curcumin possess notable antioxidant activity but are found to be significantly toxic. In this study, a novel skeleton of the monoenone monocarbonyl curcumin analogue sAc possessing reduced toxicity and improved stability was designed on the basis of the DMCA skeleton. Moreover, 32 sAc analogs were obtained by applying a green, simple, and economical synthetic method. Multiple sAc analogs with an antioxidant protective effect in PC12 cells were screened using an H2O2-induced oxidative stress damage model, and quantitative evaluation of structure–activity relationship (QSAR) model with regression coefficient of R2 = 0.918921 was built through random forest algorithm (RF). Among these compounds, the optimally active compound sAc15 elicited a potent protective effect on cell growth of PC12 cells by effectively eliminating ROS generation in response to oxidative stress injury by activating the Nrf2/HO-1 antioxidant signaling pathway. In addition, sAc15 exhibited good protection against CIRI in the mice middle cerebral artery occlusion (MCAO) model. In this paper, we provide a novel class of antioxidants and a potential compound for stroke treatment.

Protective Effects and Mechanism of Hyperoside in PC12 Cells Against Oxidative Stress Injury Induced by Hydrogen Peroxide

As the aging phenomenon continues to increase, the incidence of neurodegenerative diseases continues to increase annually. As one of the significant contributive factors of neurodegenerative diseases, oxidative stress damage has received extensive attention in recent years. Oxidative stress plays an important role in neuronal damage through various apoptotic mechanisms related to neurodegenerative diseases. The use of natural antioxidants to combat oxidative stress may be a useful approach in delaying disease progression. In this study, we explored the neuroprotective effect of hyperoside on rat pheochromoma (PC12) cells. Specifically, the antioxidant effect and mechanism of hyperoside in hydrogen peroxide (H2O2)-induced cellular cytotoxicity were investigated. Our results showed that hyperoside could significantly increase the survival rate of rat PC12 cells when exposed to H2O2. In addition, hyperoside regulated the expression of genes and proteins in the corresponding pathways by up-regulating the phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), and light chain 3β (LC3B) pathways and down-regulating the nuclear factor-ᴋ-gene binding (NF-κB), Bcl2-associated X (Bax), cysteinyl aspartate specific proteinase 3 (Caspase 3), and P62 pathways, thereby inhibiting cell apoptosis. Therefore, hyperoside can effectively inhibit H2O2-induced oxidative stress damage by regulating inflammation, autophagy, and apoptosis-related pathways.

Methylated Metabolites of Chicoric Acid Ameliorate Hydrogen Peroxide (H2O2)-Induced Oxidative Stress in HepG2 Cells.

Chicoric acid (CA) can display health benefits as a dietary polyphenol. However, as CA is widely metabolized in vivo, the actual compounds responsible for its bioactivities are not entirely known. Herein, the major methylated metabolites of CA were isolated from an in vitro co-incubation system, and their structures were elucidated. The antioxidant activities of the monomethylated metabolites (M1) and dimethylated metabolites (M2) of CA were evaluated against H2O2-induced oxidative stress damage in HepG2 cells and compared to CA. The results indicated that both M1 and M2 had better antioxidant capacities than CA by increasing cell viability, improving mitochondrial function, and balancing cellular redox status. These compounds also prevented oxidative stress by mediating the Keap1/Nrf2 transcriptional pathway and downregulating enzyme activity. The current research indicates that the methylated metabolites of CA could potentially be the candidates that are responsible for the biological efficacies attributed to CA.

Protective effect of procyanidin A-type dimers against H2O2-induced oxidative stress in prostate DU145 cells through the MAPKs signaling pathway

It has been reported that B-type procyanidins can alleviate oxidative damage of prostatic cells, but there has been limited information on the similar role of A-type procyanidins. This study investigated the protective effect of procyanidin A-type dimers from peanut skin against H2O2-induced oxidative stress damage in prostate cancer DU145 cells. According to the UPLC-Q-TOF-MS/MS analysis and comparison with standards, the fourth fraction of peanut skin procyanidin (PSP-4) was identified as procyanidin A-type dimers, namely, procyanidin A1 and A2. Results revealed that PSP-4 treatment prior H2O2 exposure increased cell activity and attenuated the cell cycle arrest and apoptosis rate. The H2O2-induced increase in intracellular reactive oxygen species (ROS) was remarkably inhibited by PSP-4. PSP-4 treatment enhanced the activity of catalase (CAT) and total super oxide dismutase (T-SOD) and restored glutathione (GSH) content, compared with the H2O2 treatment. Furthermore, the results indicated that PSP-4 protected DU145 cells by attenuating phosphorylation of the mitogen-activated protein kinases (MAPKs), by increasing the Bcl-2/Bax ratio, and by reducing the activation of caspase-3 and caspase-9 by cascade reactions. This study reveals that procyanidin A-type dimers from peanut skin have the potential function in preventing oxidative stress damage of prostatic cells.

Nanoliposomes codelivering bioactive peptides produce enhanced anti-aging effect in human skin

The present study aimed to develop nanoliposomes codelivering multiple bioactive peptides to achieve improved and comprehensive anti-aging effects in human skin. The nanoliposomes were simultaneously loaded with carnosine, palmitoyl tripeptide-5 and acetyl hexapeptide-3 (CPA-NLPs) for topical delivery. The CPA-NLPs were nano-scaled with narrow size distribution, and showed high encapsulation efficiency, high loading capacity and sustained release of bioactive peptides. Flow cytometry and confocal laser scanning microscopy showed that the nanoliposomes significantly enhanced the cellular uptake of the loaded cargo by human skin fibroblasts (HSF) cells. The CPA-NLPs also protected HSF cells from H2O2-induced oxidative stress damage and increased their expression levels of type Ι collagen and hyaluronic acid. In addition, treatment with CPA-NLPs inhibited catecholamine release in rat adrenal pheochromocytoma (PC12) cells. In a clinical trial, application of a lotion containing 2% CPA-NLPs in face led to a remarkable reduction of the volume, area and area ratio of wrinkles, and an improvement of skin elasticity. These results suggest that loading a combination of bioactive peptides into nanoliposomes is a promising strategy to enhance the topical delivery efficiency and the anti-aging efficacy.

RETRACTED ARTICLE: Chinese medicine Di-Huang-Yi-Zhi protects PC12 cells from H2O2-induced apoptosis by regulating ROS-ASK1-JNK/p38 MAPK signaling

BackgroundOxidative stress mediates the nerve injury during the pathogenesis of Alzheimer’s disease (AD). Protecting against oxidative stress damage is an important strategy to prevent and treat AD. Di-Huang-Yi-Zhi (DHYZ) is a Chinese medicine used for the treatment of AD, but its mechanism remains unknown. This study is aimed to investigate the effect of DHYZ on H2O2 induced oxidative damage in PC12 cells.MethodsPC12 cells were treated with H2O2 and DHYZ. Cell proliferation was detected by Cell counting kit-8 (CCK-8) assay. Cytotoxicity of H2O2 was measured by lactate dehydrogenase (LDH) release assay. Apoptosis were identified by Annexin V-FITC/PI staining. Caspase 3 activity was detected by commercial kit. Mitochondrial membrane potential (MMP) was detected by JC-1 staining. Reactive oxygen species (ROS) was 2′, 7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) staining. Protein expression and phosphorylation was identified by western blot.ResultsThe results showed that DHYZ antagonized H2O2-mediated cytotoxicity and proliferation inhibition. DHYZ reduced ROS production, stabilize mitochondrial membrane potential, inhibit Caspase-3 activity and apoptosis induced by H2O2. In addition, DHYZ inhibited the phosphorylation of ASK1, JNK1/2/3 and p38 MAPK which were up-regulated by H2O2.ConclusionsThe present study suggested that DHYZ protected PC12 cells from H2O2-induced oxidative stress damage and was related to inhibition of ROS production and ASK1-JNK/p38 MAPK signaling. The present study provides experimental evidence for the application of DHYZ for the management of oxidative stress damage and AD.

Neuroprotective Effects of Cornus officinalis on Stress-Induced Hippocampal Deficits in Rats and H2O2-Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells.

Oxidative stress plays a vital role in neurodegenerative diseases. Cornus officinalis (CC) has a wide range of pharmacological activities (e.g., antioxidant, neuroprotective, and anti-inflammatory). The present study was undertaken to elucidate the neuroprotective mechanism of CC and fermented CC (FCC) on stress and H2O2-induced oxidative stress damage in rats and SH-SY5Y cells. A dose of 100 mg/kg CC or FCC was orally administered to rats 1 h prior to immobilization 2 h per day for 14 days. CC, especially FCC administration decreased immobility time in forced swim test (FST), effectively alleviated the oxidative stress, and remarkably decreased corticosterone, β-endorphin and increased serotonin levels, respectively. In cells, CC and FCC significantly inhibited reactive oxygen species (ROS) generation, lactate dehydrogenase (LDH) release and significantly increased the genes expression of antioxidant and neuronal markers, such as superoxide dismutase (SOD), catalase (CAT), and brain-derived neurotrophic factor (BDNF). Moreover, the pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2 (Bax/Bcl-2) ratio was regulated by CC and FCC pretreatment. Both in rats and cells, CC and FCC downregulated mitogen-activated protein kinase (MAPK) phosphorylation. Taken together, these results demonstrated that CC and particularly FCC ameliorated oxidative stress and may be used on the neuroprotection.