Polygonum viviparum L. inhibits the lipopolysaccharide‐induced inflammatory response in RAW264.7 macrophages through haem oxygenase‐1 induction and activation of the Nrf2 pathway

BACKGROUND: Polygonum viviparum L. (PV) is a member of the family Polygonaceae and is widely distributed in high‐elevation areas. It is used as a folk remedy to treat inflammation‐related diseases. This study was focused on the anti‐inflammatory response of PV against lipopolysaccharide (LPS)‐induced inflammation in RAW264.7 macrophages. RESULTS: Treatment with PV did not cause cytotoxicity at 0–50 µg mL^?1 in RAW264.7 macrophages, and the IC₅₀ value was 270 µg mL^?1. PV inhibited LPS‐stimulated nitric oxide (NO), prostaglandin (PG)E₂, interleukin (IL)‐1β and tumour necrosis factor (TNF)‐α release and inducible NO synthase (iNOS) and cyclooxygenase (COX)‐2 protein expression. In addition, PV suppressed the LPS‐induced p65 expression of nuclear factor (NF)‐κB, which is associated with the inhibition of IκB‐α degradation. These results suggest that, among mechanisms of the anti‐inflammatory response, PV inhibits the production of NO and these cytokines by down‐regulating iNOS and COX‐2 gene expression. Furthermore, PV can induce haem oxygenase (HO)‐1 protein expression through nuclear factor E2‐related factor 2 (Nrf2) activation. A specific inhibitor of HO‐1, zinc(II) protoporphyrin IX, inhibited the suppression of iNOS and COX‐2 expression by PV. CONCLUSION: These results suggest that PV possesses anti‐inflammatory actions in macrophages and works through a novel mechanism involving Nrf2 actions and HO‐1. Thus PV could be considered for application as a potential therapeutic approach for inflammation‐associated disorders. © 2012 Society of Chemical Industry     

Anti‐inflammatory Potential of Quercetin‐3‐O‐β‐D‐(“2”‐galloyl)‐glucopyranoside and Quercetin Isolated from Diospyros kaki calyx via Suppression of MAP Signaling Molecules in LPS‐induced RAW 264.7 Macrophages

Diospyros kaki (DK) contains an abundance of flavonoids and has been used in folk medicine in Korea for centuries. Here, we report for the first time the anti‐inflammatory activities of Quercetin (QCT) and Quercetin 3‐O‐β‐(“2”‐galloyl)‐glucopyranoside (Q32G) isolated from DK. We have determine the no cytotoxicity of Q32G and QCT against RAW 264.7 cells up to concentration of 50 μM. QCT and Q32G demonstrated potent anti‐inflammatory activities by reducing expression of nitric oxide (NO), tumor necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐6 inducible NO synthase (iNOS), cyclooxygenase (COX)‐2, and mitogen‐activated protein kinase (MAPKs) in mouse RAW 264.7 macrophages activated with lipopolysaccharide (LPS). Both QCT or Q32G could decrease cellular protein levels of COX‐2 and iNOS as well as secreted protein levels of NO, PGE₂, and cytokines (TNF‐α, IL‐1β, and IL‐6) in culture medium of LPS‐stimulated RAW 264.7 macrophages. Immunoblot analysis showed that QCT and Q32G suppressed LPS‐induced MAP kinase pathway proteins p‐p38, ERK, and JNK. This study revealed that QCT and Q32G have anti‐inflammatory potential, however Q32G possess comparable activity as that of QCT and could be use as adjuvant to treat inflammatory diseases.     

Protective effect of dried safflower petal aqueous extract and its main constituent,carthamus yellow,against lipopolysaccharide‐induced inflammation in RAW264.7 macrophages

BACKGROUND: Safflower, whose botanic name is Carthamus tinctorius L., is a member of the family Compositae or Asteraceae. Carthamus yellow (CY) is the main constituent of safflower and is composed of safflomin A and safflomin B. Dried safflower petals are used in folk medicine and have been shown to invigorate blood circulation, break up blood stasis, and promote menstruation. In addition, dried safflower petals contain yellow dyes that are used to color food and cosmetics. In this study, we investigated the effects of dried safflower petals aqueous extracts (SFA) and CY on lipopolysaccharide (LPS)‐induced inflammation using RAW264.7 macrophages. RESULTS: Our data showed that treatment with SFA (1–1000 µg mL^?1) and CY (1–2000 µg mL^?1) does not cause cytotoxicity in cells. SFA and CY inhibited LPS‐stimulated nitric oxide (NO), prostaglandin E₂ (PGE₂), and interleukin 1β (IL‐1β) release, through attenuation of inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) protein expression. Further, SFA and CY suppressed the LPS‐induced phosphorylation of nuclear factor‐κB, which was associated with the inhibition of IκB‐α degradation. CONCLUSION: These results suggest that SFA and CY provide an anti‐inflammatory response through inhibiting the production of NO and PGE₂ by the downregulation of iNOS and COX‐2 gene expression. Thus safflower petals have the potential to provide a therapeutic approach to inflammation‐associated disorders. Copyright © 2010 Society of Chemical Industry     

Immunomodulatory properties of the milk whey products obtained by enzymatic and microbial hydrolysis

The objective of this study was to investigate the in vitro immunomodulatory effect of milk whey through enzymatic hydrolysis, microbial fermentation and two‐stage hydrolytic reactions (enzymatic and microbial reactions) by analysing cytokine profiles. Results indicated that the milk whey sample fermented by Lactobacillus kefiranofaciens M1 and two‐stage hydrolytic reactions (hydrolysed by Alcalase and then fermented by L. kefiranofaciens M1) could significantly induce the production of tumour necrosis factor (TNF)‐α and IL‐12 compared with the milk whey control and enzymatic treatments. Further characterisation of the immunomodulatory factor by membrane filtration and mutanolysin hydrolysation, the stimulatory activity for IL‐12 and TNF‐α production was found to be reduced and to be correlated positively with the cell wall components in L. kefiranofaciens M1. In addition, Th2‐polarised splenocytes revealed that L. kefiranofaciens M1 had both IL‐12 inducing and IL‐4 repressing activities. These results suggested that L. kefiranofaciens M1 could direct the Th1/Th2 balance toward Th1.     

Effect of bovine colostrum on the growth inhibition and differentiation of human leukemic U937 cells

BACKGROUND: Colostrum is the breast milk of female mammals produced within in a short time after giving birth. It is thought to protect neonates from infection as well as to facilitate immune system maturation. In this study the effect of bovine colostrum on the proliferation and differentiation of human leukemic U937 cells was investigated to understand more about its immunomodulatory activity. RESULTS: Human mononuclear cells (MNCs) were stimulated with bovine colostrum (CS) and then filtered to obtain a conditioned medium (CM) (CS‐MNC‐CM). CS‐MNC‐CMs prepared with day 1 to day 4 colostrums inhibited U937 cells by 39.4–64.4%. The expressions of surface markers CD11b and CD14 on U937 cells in the treated groups were 30.6–33.5% and 40.0–42.6% respectively. High levels of cytokines IL‐1β, IFN‐γ and TNF‐α were detected in CS‐MNC‐CMs. CONCLUSION: Evidence indicates that colostrums stimulate human MNCs to secrete cytokines IL‐1β, IFN‐γ and TNF‐α which subsequently inhibit the growth of U937 cells and induce their differentiation into mature monocytes and macrophages. There is a possibility for bovine colostrum to be processed into an anti‐leukemia ingredient for use in health foods. Copyright © 2009 Society of Chemical Industry     

Phenethyl isothiocyanate suppresses nitric oxide production via inhibition of phosphoinositide 3‐kinase/Akt‐induced IFN‐γ secretion in LPS‐activated peritoneal macrophages

Phenethyl isothiocyanate (PEITC), a constituent of many cruciferous vegetables, is well known to have versatile physiological activities, including chemopreventive effects. On the other hand, its anti‐inflammatory effects are poorly reported. Nitric oxide (NO) is associated with a wide variety of inflammatory diseases. In this study, we investigated the effects of PEITC on NO production in LPS‐activated peritoneal macrophages from ICR mice. The signaling pathway of LPS‐induced NO production was examined using neutralizing antibodies [anti‐interferon (IFN)‐γ and anti‐interleukin (IL‐12)] and specific protein kinase inhibitors, as well as others. The activity of PEITC toward NOx production was assessed in mice that received LPS via intraperitoneal administration. The neutralizing antibody of anti‐IFN‐γ, but not anti‐IL‐12, suppressed LPS‐induced NO production by 90%. LY294002, a specific inhibitor of phosphoinositide‐3‐kinase, suppressed Akt and IFN‐γ mRNA expression up‐regulated by LPS, whereas PEITC exhibited a similar inhibition profile. Furthermore, oral administration of PEITC significantly suppressed the serum concentration of NOx in ICR mice. Our results suggest that PEITC suppresses LPS‐induced NO production via inhibition of Akt activation and the resultant decrease in expression of IFN‐γ. This is one of the first reports to demonstrate a marked anti‐inflammatory effect of PEITC following its oral administration.     

New protein PCiP from edible golden oyster mushroom (Pleurotus citrinopileatus) activating murine macrophages and splenocytes

A new immunomodulatory protein (PCiP) was purified from an edible golden oyster mushroom (Pleurotus citrinopileatus) by extraction with 5% (v/v) cold acetic acid in the presence of 0.1% (v/v) 2‐mercaptoethanol, followed by ammonium sulfate fractionation, DE‐52 and MonoQ anion‐exchange chromatography. Electrophoresis assays demonstrated that the molecular mass of PCiP was approximately 15.0 kDa and its pI was around 5.2. PCiP is a simple protein without carbohydrate, and cannot agglutinate mouse red blood cells, suggesting PCiP is not a lectin. In addition, PCiP (5–20 µg mL^?1) alone activated murine splenocytes, and markedly increased their proliferation and gamma‐interferon (IFN‐γ secretion, but suppressed MTT metabolization, while murine splenocytes were simultaneously stimulated by the mitogen concanavalin A (ConA). Furthermore, PCiP (5–20 µg mL^?1) directly activated murine macrophages and increased the production of both the nitric oxide (NO) and tumor necrosis factor‐alpha (TNF‐α by RAW 264.7 macrophages. These findings suggest that PCiP could strengthen both the innate and adaptive responses of its host. Copyright © 2007 Society of Chemical Industry     

The inhibition of the viability of myeloma cells and the production of cytokine by two strains of Lactobacillus sakei from meat

Ten strains of lactic acid bacteria were isolated from a dry fermented sausage and tested for stimulation or inhibition of the viability of Vero and myeloma cells. They did not significantly affect the viability of Vero cells but two isolates (CBL/H and CBL/K) showed a strong and moderate inhibition of the myeloma cell viability (at 10⁸ CFU mL^?1, 17.6 and 33.2%, respectively, survival of myeloma cells). The isolates were identified as Lactobacillus sakei by DNA sequencing of the 16S rRNA products of a polymerase chain reaction. No protective effect was found, at the concentrations used, against cytotoxicity of N‐nitrosamines. To test the effect of L. sakei CBL/H and CBL/K on cytokine production [tumour necrosis factor alpha (TNF‐α), interleukin‐1β (IL‐1β) and interleukin‐8 (IL‐8)], the human macrophage cell line (THP‐1) was cultured in the presence and absence of lipopolysaccharide (LPS). Lactobacillus sakei CBL/H and CBL/K induced IL‐1β and IL‐8 release when cells were stimulated with and without LPS. However there was TNF‐α release only in the presence of the LPS.     

Anti-inflammatory activities of aqueous extract of Mesona procumbens in experimental mice

BACKGROUND: Mesona procumbens is consumed as a herbal drink and jelly‐type dessert in Taiwan. The aim of this study was to determine the mechanism of anti‐inflammatory activities of the aqueous extract of M. procumbens (AMP) using the λ‐carrageenin (Carr)‐induced mouse paw oedema model. The fingerprint chromatogram of AMP was obtained by high‐performance liquid chromatography (HPLC) analysis. To investigate the anti‐inflammatory mechanism of AMP, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and the level of malondialdehyde (MDA) in paw oedema were monitored. Serum nitric oxide (NO), tumour necrosis factor‐α (TNF‐α) and interleukin‐1β (IL‐1β) were also evaluated. RESULTS: The fingerprint chromatogram from HPLC indicated that AMP contained protocatechuic acid, chlorogenic acid, vanillic acid and caffeic acid. In the anti‐inflammatory test, AMP decreased paw oedema after Carr administration and increased the CAT, SOD and GPx activities and decreased the MDA level in paw oedema at 5 h after Carr injection. AMP also affected the serum NO, TNF‐α and IL‐1β levels at 5 h after Carr injection. Western blotting revealed that AMP decreased the expression of Carr‐induced inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2). CONCLUSION: Mesona procumbens has the potential to provide a therapeutic approach to inflammation‐associated disorders. Copyright © 2011 Society of Chemical Industry     

Mung Bean Protein Hydrolysate Modulates the Immune Response Through NF‐κB Pathway in Lipopolysaccharide‐Stimulated RAW 264.7 Macrophages

The objective of this study was to evaluate the immunomodulatory activity of mung bean protein hydrolysate (MBPH) in lipopolysaccharide (LPS)‐induced RAW 264.7 cells and discuss the possible immune regulatory mechanism. MBPH was prepared by alcalase, trypsin, neutrase, and flavourzyme. The 3‐h alcalase‐hydrolyzed hydrolysate with a molecular weight less than 1,450 Da was selected for the immunological tests. Results showed that MBPH possessed strong suppressing activity to proinflammatory mediators in a dose‐dependent manner. Compared to the LPS alone group, MBPH (200 µg/mL) significantly reduced nitric oxide (NO), inducible nitric oxide synthase, interleukin (IL)‐6, and IL‐1β secretion levels by 52.6%, 53.2%, 48.4%, and 49.7%, respectively, in LPS‐induced macrophages. It also enhanced IL‐10 secretion from 789 to 3,678 pg/mL. MBPH blocked nuclear factor‐kappa B (NF‐κB) translocation in LPS‐induced macrophages through the prevention of IκBα phosphorylation, and this process further prevented p65 translocation into the nucleus. A possible mechanism of MBPH is that it regulated the expression of inflammatory factors via the NF‐κB pathway, thus inhibiting inflammatory reactions. The results suggested that MBPH is of application potential in the development of immunomodulatory functional food to ameliorate immunosuppression.     

Regulation of inflammation,antioxidant production,and methyl-carbon metabolism during methionine supplementation in lipopolysaccharide-challenged neonatal bovine hepatocytes

Supplementation of methionine (Met) may improve immunometabolic status, specifically during a period of inflammatory stress. The aim of the present study was to establish an inflammation model using primary neonatal bovine hepatocytes and to examine the effects of increasing concentrations of dl-Met and a maintained Met to lysine (Lys) ratio on hepatocyte inflammatory responses, antioxidant production, and Met metabolism during lipopolysaccharide (LPS) challenge. Hepatocytes isolated from 4 calves were maintained as monolayer cultures and exposed to 0, 10, or 40 µMdl-Met and 100 µM Lys (0Met100Lys, 10Met100Lys, or 40Met100Lys) or 10 µMdl-Met and 25 µM Lys (10Met25Lys). Cells were exposed to each treatment for 16 h and then challenged with either 0 or 100 ng/mL of LPS for 8 h. In the absence of LPS, glutathione (GSH) was not altered by 10Met100Lys or 10Met25Lys but was increased by 40Met100Lys. With LPS challenge, GSH concentration was decreased with 40Met100Lys and tended to be decreased with 10Met100Lys. Hepatocytes receiving 10Met100Lys treated with 100 ng/mL of LPS showed an inflammatory response with increased mRNA expression of tumor necrosis factor (TNFα), IL-6, IL-1β, and interferon gamma, which was accompanied by increased nuclear factor κB inhibitor and serum amyloid A3 mRNA. The treatment 40Met100Lys was effective for preventing the LPS-induced increase in expression of the above genes except TNFα. Similar preventative effects were observed for 10Met25Lys; however, it did not prevent the LPS-induced increase in TNFα or IL-6 mRNA. Lipopolysaccharide challenge decreased mRNA expression of key genes controlling the transmethylation and Met regeneration pathways, which was not prevented by Met supplementation. The data suggest that bovine hepatocyte cultures can be used as a biological model to study the inflammatory cascade via an LPS challenge. Supplementation of Met prevents the LPS-induced hepatocyte cytokine expression and is associated with elevated intracellular GSH concentration.     

Phosphorylation of peptidoglycan from Lactobacillus acidophilus and its immunoregulatory function

Peptidoglycan (PG) is available from a wide variety of lactic acid bacteria (LAB) and is the main structure of cell wall components. Phosphorylated modification would bring new properties such as the potential antioxidant activities and antiviral capability to an organic molecule. In the present work, small molecular fragments of PG (derived from Lactobacillus acidophilus) hydrolysed by mutanolysin were phosphorylated under optimal conditions. P‐PG had a monomer molecular structure of GlcNAC[PO₃]–MurNAC–Ala–Glu–Lys–Ala, with a molecular mass of 884 Da and a phosphorus content of 8.9%. P‐PG displayed some immunoregulatory activity in lipopolysaccharides (LPS) stimulated RAW 264.7 macrophages. Compared with the LPS‐stimulated group, the addition of P‐PG inhibited the secretion of GM‐CSF, TNF‐α and IL‐1 in a dose‐dependent manner. The effect of 50 μg mL^?1 of P‐PG was more significant than 50 μg mL^?1 of PG. Lower fluorescence of lysosomes was observed in P‐PG‐treated RAW 264.7 cells may also reveal some immune defence function in the LPS‐induced macrophages.     

Enhanced Anti‐Inflammatory Activities by the Combination of Luteolin and Tangeretin

Dietary components in combination may act synergistically and produce enhanced biological activities. Herein, we investigated the anti‐inflammatory effects of 2 flavonoids, that is luteolin (LUT) and tangeretin (TAN) in combination. Lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophages were treated with noncytotoxic concentrations of LUT, TAN, and their combinations. The results showed that LUT/TAN in combination produced synergistic inhibitory effects on LPS‐stimulated production of nitric oxide (NO). ELISA results demonstrated that LUT/TAN in combination caused stronger suppression on the LPS‐induced overexpression of proinflammatory mediators, such as prostaglandin E₂ (PGE₂), interleukin (IL)‐1β, and IL‐6 than LUT or TAN alone. Immunoblotting and Real‐Time PCR analyses showed that LUT/TAN combination significantly decreased LPS‐induced protein and mRNA expression of inducible nitric oxide synthase and cyclooxygenase‐2. These inhibitory effects of the combination treatment were stronger than those produced by LUT or TAN alone. Overall, our results demonstrated for the first time that combination of LUT and TAN produced synergistic anti‐inflammatory effects in LPS‐stimulated RAW 264.7 macrophages.     

Comparison of dipeptidyl peptidase IV-inhibitory activity of peptides from bovine and caprine milk casein by in silico and in vitro analyses

The different potential for release of dipeptidyl peptidase IV (DPP-IV)-inhibitory peptides from bovine and caprine milk casein was assessed by in silico analysis and in vitro proteolytic assays. The former predicted a weakly higher potency for caprine casein than bovine as a precursor of DPP-IV-inhibitory peptides, although with markedly diverse potential for individual caseins. This was verified by protease hydrolysis; trypsin-treated casein hydrolysates (TCAHs) displayed the strongest bioactivity. Fractionation of caprine TCAHs revealed slightly, but significantly, higher inhibitory activity in peptides <5 kDa, and notably greater efficiency in the >5 kDa fraction, than their bovine counterparts. Through in silico trypsin hydrolysis and peptides synthesis, four novel caprine casein-derived DPP-IV-inhibitory peptides, including GPFPILV and HPINHR (half maximal inhibitory concentration = 163.7 ± 1.33 and 452.2 ± 7.15 μm, respectively), were found. This study corroborates the weak superiority of caprine casein over bovine in release of DPP-IV-inhibitory peptides, and identifies several novel caprine casein-derived DPP-IV-inhibitory peptides.     

Characterization,Anti‐Inflammatory and Antiproliferative Activities of Natural and Sulfonated Exo‐Polysaccharides from Streptococcus thermophilus ASCC 1275

Exo‐polysaccharides (EPS) isolated from Streptococcus thermophilus ASCC 1275 were sulfated (31%). High‐performance liquid chromatography identified that EPS was composed of mannose (30.19%), galactose (20.10%), glucose (18.05%), glucosamine (16.04%), galactosamine (9.06%), glucuronic acid (3.55%), and ribose (3.01%). Pro‐/anti‐inflammatory cytokine secretion ratios (IL‐1β/IL‐10, IL‐6/IL‐10, and TNF‐α/IL‐10) of lipopolysaccharide stimulated RAW 264.7 macrophages were significantly decreased by EPS and S.EPS treatments in a dose dependent manner. Furthermore, anti‐inflammatory activities of S.EPS improved 49.3% and 24.0% than those of EPS before or after LPS treatment. The reactive oxygen species were inhibited by EPS and S.EPS by 49.6% and 55.1% at 50 μg/mL, respectively. Inhibition activities of S.EPS on nitric oxide production were 12.9% and 55.4% higher than those of EPS at 10 and 50 μg/mL. Additionally, S.EPS exhibited stronger antiproliferative activity on Caco‐2 and HepG2 cells. Our results indicated that anti‐inflammatory and antiproliferative activities of EPS were significantly (P < 0.01) improved by sulfonation.     

Strawberry,loquat, mulberry,and bitter melon juices exhibit prophylactic effects on LPS-induced inflammation using murine peritoneal macrophages

We hypothesized the juices from strawberry, loquat, mulberry and bitter melon exhibit anti-inflammatory activities using lipopolysaccharide (LPS)-stimulated murine peritoneal macrophage cultures. Selected juices were administered as a prophylactic, postmortem or concurrent event relative to LPS stimulation to clarify the effective mechanisms. Selected fruits and vegetable juices were administered to macrophage cultures for 24 h prior to LPS stimulation (model A). Selected samples were administered to cell cultures at 24 h following LPS treatment (model B). Selected fruits and vegetable juices and LPS were simultaneously co-cultured with macrophages for 24 h (model C). The LPS-induced secretions of pro-inflammatory cytokines interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α and anti-inflammatory cytokine IL-10 were determined. The results showed that strawberry, loquat, mulberry and bitter melon administration increased IL-10 production by LPS-stimulated peritoneal macrophages in dose-dependent manners in experimental model A. Simultaneously, loquat, mulberry, and bitter melon administrations significantly (P < 0.05) decreased the levels of IL-1β, IL-6 and/or TNF-α. Administration with loquat and bitter melon to experimental model C significantly increased IL-10 production. This study suggests that strawberry, loquat, mulberry, and bitter melon juices exhibit a prophylactic effect on LPS-induced inflammation of peritoneal macrophages via increasing anti-inflammatory cytokine and/or decreasing pro-inflammatory cytokines secretions.     

EFFECTS OF CONJUGATED LINOLEIC ACID ISOMERS ON SERUM TUMOR NECROSIS FACTOR-A CONCENTRATION IN MICE

In previously studies, we showed that tumor necrosis factor‐α (TNF‐α), a cytokine involved in a variety of biologically important events, is partially involved in the effects of conjugated linoleic acid (CLA). In this report, we further tested the effects of individual CLA isomers on serum TNF‐α concentration along with other biological markers in mice. The animals were fed experimental diets (control, 0.5% CLA‐mixed isomer, 0.25% cis‐9,trans‐11 CLA or 0.25% trans‐10,cis‐12 CLA) for 5 days and were then challenged with endotoxin. Both cis‐9,trans‐11 and trans‐10,cis‐12 CLA isomers reduced serum TNF‐α levels compared to control. CLA had no effect on the other biological markers examined. These results suggest possible early involvement of CLA in immune and/or inflammatory responses, followed by reduction of body fat. Its effect on TNF‐α helps explain in part how CLA modulates other biological functions such as immune response, insulin responses, atherosclerosis and cancer.     

Antioxidative and Anti-Inflammatory Neuroprotective Effects of Astaxanthin and Canthaxanthin in Nerve Growth Factor Differentiated PC12 Cells

ABSTRACT: Nerve growth factor differentiated PC12 cells were used to examine the antioxidative and anti‐inflammatory effects of astaxanthin (AX) and canthaxanthin (CX). PC12 cells were pretreated with AX or CX at 10 or 20 μM, and followed by exposure of hydrogen peroxide (H₂O₂) or 1‐methyl‐4‐phenylpyridinium ion (MPP⁺) to induce cell injury. H₂O₂ or MPP⁺ treatment significantly decreased cell viability, increased lactate dehydrogenase (LDH) release, enhanced DNA fragmentation, and lowered mitochondrial membrane potential (MMP) (P < 0.05). The pretreatments from AX or CX concentration‐dependently alleviated H₂O₂ or MPP⁺‐induced cell death, LDH release, DNA fragmentation, and MMP reduction (P < 0.05). Either H₂O₂ or MPP⁺ treatment significantly increased malonyldialdehyde (MDA) and reactive oxygen species (ROS) formations, decreased glutathione content, and lowered glutathione peroxidase (GPX) and catalase activities (P < 0.05). The pretreatments from AX or CX significantly retained GPX and catalase activities, and decreased MDA and ROS formations (P < 0.05). H₂O₂ or MPP⁺ treatment significantly decreased Na⁺‐K⁺‐ATPase activity, elevated caspase‐3 activity and levels of interleukin (IL)‐1, IL‐6, and tumor necrosis factor (TNF)‐α (P < 0.05); and the pretreatments from these agents significantly restored Na⁺‐K⁺‐ATPase activity, suppressed caspase‐3 activity and release of IL‐1, IL‐6, and TNF‐α (P < 0.05). Based on the observed antioxidative and anti‐inflammatory protection from AX and CX, these 2 compounds were potent agents against neurodegenerative disorder.