Rutin attenuates vancomycin‐induced renal tubular cell apoptosis via suppression of apoptosis,mitochondrial dysfunction,and oxidative stress

Vancomycin is a glycopeptide antibiotic widely used to treat infections caused by methicillin‐resistant Staphylococcus aureus. However, nephrotoxicity is a major adverse side effect, and the development of effective nephroprotective agents remains a priority in antimicrobial chemotherapy. In this study, we investigated the cell protective effects of the flavonol glycoside rutin against vancomycin‐induced toxicity. Vancomycin added to porcine renal tubular LLC‐PK1 cells caused an increase of production of intracellular reactive oxygen species and subsequent apoptotic cell death. Pretreatment of LLC‐PK1 cells with rutin at 5, 10, and 20 μM for 2 hr prior to 2‐mM vancomycin exposure for 24 hr significantly decreased intracellular reactive oxygen species and increased superoxide dismutase and catalase activities. Rutin pretreatment also protected cells from vancomycin‐induced caspase activation, mitochondrial membrane depolarization, and subsequent apoptosis. This study demonstrates a protective effect of rutin and suggests that rutin coadministration is an alternative therapy for treatment of vancomycin‐induced nephrotoxicity.     

Extracts from Calendula officinalis Offer in Vitro Protection Against H2O2 Induced Oxidative Stress Cell Killing of Human Skin Cells

The in vitro safety and antioxidant potential of Calendula officinalis flower head extracts was investigated. The effect of different concentrations (0.125, 0.5, 1.0, 2.0 and 5.0% (v/v)) of Calendula extracts on human skin cells HaCaT in vitro was explored. Doses of 1.0% (v/v) (0.88 mg dry weight/mL) or less showed no toxicity. Cells were also exposed to the Calendula extracts for either 4, 24 or 48 h before being exposed to an oxidative insult (hydrogen peroxide H₂O₂) for 1 h. Using the MTT cytotoxicity assay, it was observed that two independent extracts of C. officinalis gave time‐dependent and concentration‐dependent H₂O₂ protection against induced oxidative stress in vitro using human skin cells. Pre‐incubation with the Calendula extracts for 24 and 48 h increased survival relative to the population without extract by 20% and 40% respectively following oxidative challenge. The antioxidant potential of the Calendula extracts was confirmed using a complimentary chemical technique, the DPPH^● assay. Calendula extracts exhibited free radical scavenging abilities. This study demonstrates that Calendula flower extracts contain bioactive and free radical scavenging compounds that significantly protect against oxidative stress in a human skin cell culture model. Copyright © 2014 John Wiley & Sons, Ltd.     

Carthamus tinctorius flower extract prevents H2O2‐induced dysfunction and oxidative damage in osteoblastic MC3T3‐E1 cells

The flowers of Carthamus tinctorius L. (Compositae) have been widely used for enhancing blood circulation and postmenopausal disorder in women. In the present study, the potential protective effects of C. tinctorius flower extract (CFE) against reactive oxygen species (ROS) induced osteoblast dysfunction were investigated using osteoblastic MC3T3‐E1 cells. The osteoblast function was assessed by measuring alkaline phosphatase activity, collagen content, calcium deposition, and RANKL production, and the oxidative status was assessed by measuring intracellular lipid peroxidation, and protein oxidation in osteoblastic MC3T3‐E1 cells. A significant reduction in the alkaline phosphatase activity, collagen, and calcium deposition and an increase in the production of receptor activator of nuclear factor‐kB ligand (RANKL) were observed after 0.3 mM H₂O₂ addition. The H₂O₂‐induced alterations were prevented by pre‐incubating the osteoblasts with 2–10 μg/ml CFE for 48 h. When the oxidative stress was induced by H₂O₂, the increased production of protein carbonyl and malondialdehyde was also reduced at the same CFE concentration. These results demonstrate that C. tinctorius flower can act as a biological antioxidant in a cell culture experimental model and protect osteoblasts from oxidative stress‐induced toxicity. Copyright © 2009 John Wiley & Sons, Ltd.     

Effect of Cinnamoyl and Flavonol Glucosides Derived from Cherry Blossom Flowers on the Production of Advanced Glycation End Products (AGEs) and AGE‐induced Fibroblast Apoptosis

Cherry blossom flowers are familiar to the Japanese, and some species of the flowers soaked in salty vinegar are used as processed foods. The constituents of aqueous ethanol extract from cherry blossom (Prunus lannesiana) flowers (CBE) were examined and cinnamoyl and flavonol glucosides were isolated. To elucidate the pharmacological functions of CBE and its constituents, their effects on the production of advanced glycation end products (AGEs) and on AGE‐induced fibroblast damage were examined. CBE and 1‐O‐(E)‐caffeoyl‐β‐d ‐glucopyranoside (CaG), a principal compound in CBE, significantly suppressed the production of AGEs derived from glucose and albumin at 100 μg/mL. Among the flavonol glucosides, quercetin 3‐O‐β‐d ‐glucopyranoside (QG) exhibited potent suppressive activity (IC₅₀: 30 μg/mL). CBE and CaG suppressed glyoxal‐induced AGE production in fibroblasts at 10 μg/mL, but QG did not. In addition, CBE and CaG recovered collagen lattice formation consisting of collagen and glycated fibroblasts at 10 μg/mL. Moreover, CBE and its constituents, except kaempferol 3‐O‐(6″‐malony)‐β‐d ‐glucopyranoside, significantly suppressed fibroblast apoptosis induced by carboxymethyl lysine‐collagen at 10 μg/mL. These results show that cinnamoyl and flavonol glucosides of cherry blossom flowers suppress AGE production and AGE‐induced fibroblast apoptosis. Cherry blossom flowers may be effective against skin AGE production and fibroblast damage by AGEs. Copyright © 2011 John Wiley & Sons, Ltd.     

In vitro protective effects of Withania somnifera (L.) dunal root extract against hydrogen peroxide and β‐amyloid(1–42)‐induced cytotoxicity in differentiated PC12 cells

Withania somnifera L. Dunal (Solanaceae), also known as ‘ashwagandha’ in Sanskrit and as ‘Indian ginseng’, is used widely in Ayurvedic medicine as a nerve tonic and memory enhancer, with antiaging, antistress, immunomodulatory and antioxidant properties. There is a paucity of data on the potential neuroprotective effects of W. somnifera root, as traditionally used, against H₂O₂‐ and Aβ_(1–42)‐induced cytotoxicity which are current targets for novel approaches to treat dementia, especially dementia of the Alzheimer's type (AD). In this study, an aqueous extract prepared from the dried roots of W. somnifera was assessed for potential protective effects against H₂O₂‐ and Aβ_(1–42)‐aggregated fibril cytotoxicity by an MTT assay using a differentiated rat pheochromocytoma PC12 cell line. The results suggest that pretreatments of differentiated PC12 cells with aqueous extracts of W. somnifera root significantly protect differentiated PC12 cells against both H₂O₂‐ and Aβ_(1–42)‐induced cytotoxicity, in a concentration dependent manner. To investigate the compounds that could explain the observed effects, the W. somnifera extract was analysed by liquid chromatography–serial mass spectrometry and numerous withanolide derivatives, including withaferin A, were detected. These results demonstrate the neuroprotective properties of an aqueous extract of W. somnifera root and may provide some explanation for the putative ethnopharmacological uses of W. somnifera for cognitive and other neurodegenerative disorders that are associated with oxidative stress. Copyright © 2010 John Wiley & Sons, Ltd.     

Bacopa monnieri‐Induced Protective Autophagy Inhibits Benzo[a]pyrene‐Mediated Apoptosis

Benzo[a]pyrene (B[a]P) is capable of inducing oxidative stress and cellular injuries leading to cell death and associates with a significant risk of cancer development. Prevention of B[a]P‐induced cellular toxicity with herbal compound through regulation of mitochondrial oxidative stress might protect cell death and have therapeutic benefit to human health. In this study, we demonstrated the cytoprotective role of Bacopa monnieri (BM) against B[a]P‐induced apoptosis through autophagy induction. Pretreatment with BM rescued the reduction in cell viability in B[a]P‐treated human keratinocytes (HaCaT) cells indicating the cytoprotective potential of BM against B[a]P. Moreover, BM was found to inhibit B[a]P‐mediated reactive oxygen species (ROS)‐induced apoptosis activation in HaCaT cells. Furthermore, BM was found to preserve mitochondrial membrane potential and inhibited release of cytochrome c in B[a]P‐treated HaCaT cells. Bacopa monnieri induced protective autophagy; we knocked down Beclin‐1, and data showed that BM was unable to protect from B[a]P‐induced mitochondrial ROS‐mediated apoptosis in Beclin‐1‐deficient HaCaT cells. Moreover, we established that B[a]P‐induced damaged mitochondria were found to colocalize and degraded within autolysosomes in order to protect HaCaT cells from mitochondrial injury. In conclusion, B[a]P‐induced apoptosis was rescued by BM treatment and provided cytoprotection through Beclin‐1‐dependent autophagy activation. Copyright © 2016 John Wiley & Sons, Ltd.     

Inhibition of UVB‐induced pro‐inflammatory cytokines and MMP expression by Zanthoxylum rhetsa bark extract and its active constituent hesperidin

The antiphoto aging property of Zanthoxylum rhetsa obtained from Pangkor Island, Malaysia, was evaluated. Solvent fractions of different polarity obtained from the methanolic extract of the bark material were initially tested for anticollagenase and antielastase activities. The ethyl acetate fraction showed bioactivity against the protease enzymes. Hence, it was subjected to further purification via column chromatography, to yield a major constituent, hesperidin. Subsequently, the ethyl acetate fraction and hesperidin were tested for their effects against UVB‐induced cytotoxicity and expressions of inflammatory cytokines (IL‐6, IL‐1β, and TNF‐α), NF‐κB, and MMPs (MMP1, 3, and 9) in human dermal fibroblasts (HDF). Both fraction and pure compound prevented UVB‐induced cytotoxicity in HDF cells, in a dose dependent manner. Moreover, the ethyl acetate fraction inhibited the increase of pro‐inflammatory cytokines induced by UVB to a level similar to the control (without UV treatment). Additionally, the fraction significantly inhibited the expressions of NF‐κB, MMP 1, MMP 3, and MMP 9 in HDF cells treated with UVB. Similar effects were observed with hesperidin. The results obtained suggested that the ethyl acetate fraction of Z. rhetsa and its bioactive constituent, hesperidin, have the potential to be used as active ingredients in sunscreen and antiphoto aging formulations.     

Photoprotective Activity of Vulpinic and Gyrophoric Acids Toward Ultraviolet B‐Induced Damage in Human Keratinocytes

Vulpinic and gyrophoric acids are known as ultraviolet filters for natural lichen populations because of their chemical structures. However, to the best of our knowledge, there has been no reference to their cosmetic potential for skin protection against ultraviolet B (UVB)‐induced damage and, consequently, we propose to highlight their photoprotective profiles in human keratinocytes (HaCaT). Therefore, vulpinic acid and gyrophoric acid were isolated from acetone extracts of Letharia vulpina and Xanthoparmelia pokornyi, respectively. Their photoprotective activities on irradiated HaCaT cells and destructive effects on non‐irradiated HaCaT cells were compared through in vitro experimentation: 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide and lactate dehydrogenase assays, 4′,6‐diamino‐2‐phenylindole and tetramethylrhodamine B isothiocyanate‐phalloidin staining protocols. Both of the lichen substances effectively prevented cytotoxic, apoptotic and cytoskeleton alterative activities of 2.5 J/cm² UVB in a dose‐dependent manner. Moreover, vulpinic and gyrophoric acids showed no toxic, apoptotic or cytoskeleton alterative effects on non‐irradiated HaCaT cells, except at high doses (≥400 μM) of gyrophoric acid. The findings suggest that vulpinic and gyrophoric acids can be promising cosmetic ingredients to photo‐protect human skin cells and should therefore be further investigated by in vitro and in vivo multiple bioassays. Copyright © 2015 John Wiley & Sons, Ltd.     

Antiproliferation of keratinocytes and alleviation of psoriasis by the ethanol extract of Artemisia capillaris

The therapeutic potentials of the ethanol extract of Artemisia capillaris (ACE) for psoriasis were verified in HaCaT cells (as an immortalized human keratinocyte cell line) and imiquimod (IMQ)‐induced psoriasis‐like mouse models. In HaCaT cells, IC₅₀ value of ACE was 37.5 μg/ml after incubating for 72 hr. The antiproliferation activity of ACE in HaCaT cells was further verified by apoptosis assays. The percentage of apoptotic population in ACE‐treated group was significantly higher than that of control group (p < .05). The result of cell cycle arrest assay also supported the observed antiproliferation efficacy of ACE in HaCaT cells. In IMQ‐induced psoriasis‐like mouse models, the Psoriasis Area and Severity Index score of ACE (50 mg/ml; ACE50)‐treated group was significantly lower than that of IMQ group on Day 4 (p < .05). After topical application of ACE on psoriasis‐like lesion for 4 days, the epidermal thickness of (IMQ + ACE50) group was significantly lower than that of IMQ group (p < .05). The expression levels of Ki‐67 and intracellular adhesion molecule‐1 in excised skin tissues of (IMQ + ACE50) group were also lower than those of IMQ group. All these findings suggest that ACE can be used as a promising antipsoriatic agent.     

Pueraria thunbergiana inhibits cisplatin‐induced damage of HEI‐OC1 auditory cells through scavenging free radicals

The radix of Pueraria thunbergiana (P. thunbergiana) is traditionally prescribed to attenuate the clinical manifestation of inner ear dysfunction and various clinical situations including fevers, gastrointestinal disorders, skin problems, migraine headaches, lowering cholesterol, and treating chronic alcoholism in oriental medicine. In the present study, we examined the protective effect of ethanol extract of the radix of P. thunbergiana (RPT) on cisplatin‐induced damage of HEI‐OC1 auditory hair cells. When the cells were cultured in the medium containing 5–100 μg/mL of RPT, RPT showed protective effect against the cisplatin‐induced HEI‐OC1 cell damage. We also measured the effects of RPT on lipid peroxidation of cisplatin‐treated cells as well as scavenging activities against superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. RPT reduced cisplatin‐induced lipid peroxidation in a dose‐dependent manner. Furthermore, RPT showed strong scavenging activity against superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. These results indicate that RPT protects cisplatin‐induced HEI‐OC1 cell damage through inhibition of lipid peroxidation and scavenging activities of free radials. Copyright © 2009 John Wiley & Sons, Ltd.     

Hepatoprotective activity of praecoxin A isolated from Melaleuca ericifolia against carbon tetrachloride‐induced hepatotoxicity in mice. Impact on oxidative stress,inflammation, and apoptosis

The hepatoprotective activity of praecoxin A, an ellagitannin from Melaleuca ericifolia, was determined against CCl₄‐induced toxicity in mice. Praecoxin A was administered (25, 50, and 100 mg/kg) for 5 days followed by CCl₄. Praecoxin A markedly ameliorated the CCl₄‐induced increase in AST (by 19, 52, and 56%), ALP (22, 45, and 48%), ALT (11, 47, and 54%), total bilirubin (14, 27, and 28%), and MDA (26, 44, and 51%) at the tested doses, respectively, as compared with CCl₄ group. It was evident that praecoxin A significantly (p < 0.001) increased the antioxidant parameters GSH (45, 99, and 137%) and SOD (61, 129, and 159%). Histological findings revealed a marked amelioration of hepatocyte degeneration, necrosis, inflammatory cell infiltration, and hemorrhage in the groups treated with praecoxin A. COX‐2 and caspase‐3 hepatic expressions were significantly downregulated (p < 0.001) in praecoxin A‐treated groups (up to 57, 83, and 93% for COX‐2 and by 30, 82, and 99% for caspase‐3). These findings suggest that praecoxin A exerts a beneficial effect against oxidative stress by reducing lipid peroxidation, enhancing the antioxidant defense status, and protecting against the histopathological changes induced by CCl₄. This study highlights a promising natural hepatoprotective candidate derived from M. ericifolia that might be an alternative to silymarin.     

Pinus densiflora bark extract ameliorates 2,4‐dinitrochlorobenzene‐induced atopic dermatitis in NC/Nga mice by regulating Th1/Th2 balance and skin barrier function

Korean red pine (Pinus densiflora) bark has been traditionally used in Korea and other parts of East Asia to relieve inflammatory diseases. Although many studies using P. densiflora bark have been reported, its effect on atopic dermatitis (AD) has not been elucidated. Thus, we investigated whether the P. densiflora bark extract (PBE) has potential to attenuate AD symptoms and elucidated the molecular mechanism. Oral administration of PBE to mice with 2,4‐dinitrochlorobenzene (DNCB)‐induced AD lessened dermatitis scores and scratching behavior and significantly reduced measures of epidermal thickness, infiltration of mast cells and eosinophils, levels of immunoglobulin E (IgE), and IgG₁/IgG_2a ratio in serum. PBE not only inhibited IL‐4, IL‐5, and IL‐13 but also increased IFN‐γ in splenic production. Furthermore, PBE significantly suppressed mRNA expression of thymic stromal lymphopoietin and further downregulated the mRNA expression of Th2 and Th17 cytokines such as IL‐4, IL‐13, IL‐17, IL‐31, and TNF‐α. In addition, the protein expressions of filaggrin, involucrin, and loricrin in lesional skin were recovered by PBE. These results suggest that PBE attenuates DNCB‐induced AD via regulating Th1/Th2 balance and skin barrier function.     

The protective effects of hydroxytyrosol against UVB‐induced DNA damage in HaCaT cells

The chemoprotective effect of hydroxytyrosol (HT) against UVB‐induced DNA damage was investigated in a human skin keratinocyte cell line, HaCaT. The comet assay was used to monitor DNA strand breaks. Intracellular reactive oxygen species (ROS) formation was measured by flow cytometry using 2,7‐dichlorofluorescein diacetate (DCFH‐DA). The levels of oxidatively generated damage to DNA were estimated by immunocytochemistry analysis of 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG). The protein expression of p53 and NF‐κB was estimated by western blotting. The results showed that HT significantly reduced the DNA strand breaks caused by UVB. It was also found that HT reduced intracellular ROS formation and 8‐OHdG level caused by UVB. Furthermore, HT attenuated the expression of p53 and NF‐κB in a concentration‐dependent manner. These results strongly suggest that HT has a significant protective ability against UVB‐induced DNA damage and that oxidative stress plays an important part in it. Copyright © 2009 John Wiley & Sons, Ltd.     

Scutellaria radix Extract as a Natural UV Protectant for Human Skin

Ultraviolet (UV) radiation induces oxidative injury and inflammation in human skin. Scutellaria radix (SR, the root of Scutellaria baicalensis Georgi) contains flavonoids with high UV absorptivity and antioxidant properties. The purpose of this study was to examine the potential use of SR extract as an additive in cosmetic products for UV protection. SR extract and its butanol (BuOH) fraction strongly absorbed UV radiation and displayed free radical scavenging activity against 2,2‐diphenyl‐1‐picrylhydrazyl radials and 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) radicals. They also attenuated the UV‐induced death of HaCaT cells. Sunscreen creams, with or without supplementation of SR extract BuOH fraction, were tested in vivo in human trials to evaluate potential skin irritation and determine the sun protection factor (SPF). Both sunscreen creams induced no skin irritation. A sunscreen cream containing 24% ZnO showed an SPF value of 17.8, and it increased to 22.7 when supplemented with 5% SR extract BuOH fraction. This study suggests that SR‐derived materials are useful as safe cosmetic additives that provide UV protection. Copyright © 2015 John Wiley & Sons, Ltd.     

The ability of hydroxysafflor yellow a to attenuate lipopolysaccharide‐induced pulmonary inflammatory injury in mice

Hydroxysafflor yellow A (HSYA) is a component of the flower of Carthamus tinctorius L. The present study investigated whether HSYA could attenuate acute lung injury (ALI) induced by lipopolysaccharide (LPS) administration. Male Kunming mice were pretreated with HSYA 0.5 h prior to intraperitoneal application of LPS. Arterial blood gas, lung water content index, lung tissue myeloperoxidase (MPO) activity, mRNA expression of inflammatory cytokines, NF‐κBp65, p38 mitogen‐activated protein kinase (MAPK) and pathological changes in lung morphology were assessed. After LPS administration, all animals displayed increased arterial carbon dioxide partial pressure (PaCO₂), and decreased arterial oxygen partial pressure (PaO₂), arterial oxygen saturation (SO₂), HCO₃^? concentration and pH, which were ameliorated by pretreating the animals with HSYA. HSYA administration significantly attenuated inflammatory cell infiltration and alleviated pulmonary edema induced by LPS. Moreover, HSYA decreased NF‐κB p65 nuclear translocation, inhibited proinflammatory cytokine TNF‐α, IL‐1β and IL‐6 mRNA expression and promoted antiinflammatory cytokine IL‐10 gene expression following LPS injection. Pulmonary p38 MAPK phosphorylation was upregulated 4 h after LPS treatment, which could be suppressed by pretreatment with HSYA. These findings demonstrated the protective effect of HSYA against LPS‐induced acute lung injury, which is suggested to be associated with the inhibition of p38 MAPK, NF‐κB p65 activation and alteration of inflammatory cytokine expression. Copyright © 2010 John Wiley & Sons, Ltd.     

Genistein inhibits Aβ25–35‐induced SH‐SY5Y cell damage by modulating the expression of apoptosis‐related proteins and Ca2+ influx through ionotropic glutamate receptors

In this study, we investigated the protective effects of genistein against SH‐SY5Y cell damage induced by β‐amyloid 25–35 peptide (Aβ_25–35) and the underlying mechanisms. Aβ‐induced neuronal death, apoptosis, glutamate receptor subunit expression, Ca²⁺ ion concentration, amino acid transmitter concentration, and apoptosis‐related factor expression were evaluated to determine the effects of genistein on Aβ‐induced neuronal death and apoptosis. The results showed that genistein increased the survival of SH‐SY5Y cells and decreased the level of apoptosis induced by Aβ_25–35. In addition, genistein reversed the Aβ_25–35‐induced changes in amino acid transmitters, α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionate (AMPA) receptors, and N‐methyl‐d ‐aspartate (NMDA) receptor subunits in SH‐SY5Y cells. Aβ_25–35‐induced changes in Ca²⁺ and B‐cell lymphoma‐2 (Bcl‐2) and Bcl‐2‐associated X (Bax) protein and gene levels in cells were also reversed by genistein. Our data suggest that genistein protects against Aβ_25–35‐induced damage in SH‐SY5Y cells, possibly by regulating the expression of apoptosis‐related proteins and Ca²⁺ influx through ionotropic glutamate receptors.     

The effect of xanthorrhizol on the expression of matrix metalloproteinase‐1 and type‐I procollagen in ultraviolet‐irradiated human skin fibroblasts

Matrix metalloproteinases (MMPs) are key regulators of the skin photoaging process that is set in motion by exposure to ultraviolet (UV) irradiation. This skin damage results from UV‐induced generation of reactive oxygen species, which are associated with upregulation of MMPs and decreased collagen synthesis. We investigated the effects of xanthorrhizol, isolated from Curcuma xanthorrhiza, on the expression of MMP‐1 and type‐I procollagen in UV‐irradiated human skin fibroblasts. Fibroblasts cultured in the presence or absence of purified xanthorrhizol or C. xanthorrhiza extract were irradiated with UV (20 mJ/cm²), and MMP‐1 and type‐I procollagen levels were measured using Western blot analysis. Xanthorrhizol (0.001–0.1 µM ) and C. xanthorrhiza extract (0.01–0.5 µg/mL) induced a significant, dose‐dependent decrease in the expression of MMP‐1 protein, and increased the expression of type‐1 procollagen. At a concentration of 0.1 µM , xanthorrhizol nearly completely abrogated MMP‐1 expression. The MMP‐1‐suppressing and type‐1 procollagen‐inducing effects of xanthorrhizol treatment were greater than those of epigallocatechin 3‐O‐gallate (EGCG), which is known to be a natural anti‐aging agent. These results suggest that xanthorrhizol is a potential candidate for the prevention and treatment of skin aging. Copyright © 2009 John Wiley & Sons, Ltd.     

Alantolactone and Isoalantolactone Prevent Amyloid β25–35‐induced Toxicity in Mouse Cortical Neurons and Scopolamine‐induced Cognitive Impairment in Mice

Given the evidence for detoxifying/antioxidant enzyme‐inducing activities by alantolactone (AL) and isoalantolactone (IAL), the purpose of this study was to investigate the effects of AL and IAL on Aβ_25–35‐induced cell death in mouse cortical neuron cells and to determine their effects on scopolamine‐induced amnesia in mice. Our data demonstrated that both compounds effectively attenuated the cytotoxicity of Aβ_25–35 (10 μM) in neuronal cells derived from the mouse cerebral cortex. It was also found that the production of intracellular reactive oxygen species, including superoxide anion induced by Aβ_25–35, was inhibited. Moreover, the administration of the sesquiterpenes reversed scopolamine‐induced cognitive impairments as assessed by Morris water, Y‐maze, and the passive avoidance tests, and the compounds decreased acetylcholinesterase (AChE) activities in a dose‐dependent manner. Interestingly, AL and IAL did not improve scopolamine‐induced cognitive deficit in Nrf2 ^?/? mice, suggesting that memory improvement by sesquiterpenes was mediated not only by the activation of the Nrf2 signaling pathway but also by their inhibitory activity against AChE. In conclusion, our results showed that AL and IAL had neuroprotective effects and reversed cognitive impairments induced by scopolamine in a mouse model. Therefore, AL and IAL deserve further study as potential therapeutic agents for reactive oxygen species‐related neurodegenerative diseases. Copyright © 2017 John Wiley & Sons, Ltd.     

Growth inhibitory activity of biflavonoids and diterpenoids from the leaves of the Libyan Juniperus phoenicea against human cancer cells

Three biflavonoids [cupressuflavone ( 1 ), amentoflavone ( 2 ), and sumaflavone ( 3 )], four diterpenoids [13‐epi‐cupressic acid ( 4 ), imbricatholic acid ( 5 ), 3‐hydroxy‐sandaracopimaric acid ( 6 ), and dehydroabietic acid ( 7 )], and one lignan [β‐peltatin methyl ether ( 8 )] were isolated from the cytotoxic fractions of the extracts of the leaves of the Libyan Juniperus phoenicea L. The structures of these compounds were elucidated by spectroscopic means. Cytotoxicity of compounds 1 – 6 were assessed against the human lung cancer cell line A549 using the MTT assay. Compounds 1 and 3 showed cytotoxicity against the A549 cells (IC₅₀ = 65 and 77 μM, respectively), whereas compound 2 did not show any activity. Diterpenes 4 – 6 exhibited weak cytotoxicity against the A549 cells with the IC₅₀ values of 159, 263, and 223 μM, respectively. The cytotoxicity of each compound was compared with the anticancer drug, etoposide (IC₅₀ = 61 μM). Cupressuflavone ( 1 ) was evaluated also for cytotoxicity against both the human PC3 cancer cell line and the normal prostate cell line (PNT2), and this compound revealed a high degree of cytotoxic selectivity towards the prostate cancer cells (PC3), with IC₅₀ value of 19.9 μM, without any evidence of cytotoxicity towards the normal prostate cell line (PNT2).     

Inhibitory Effect of Thymoquinone on Testosterone‐Induced Benign Prostatic Hyperplasia in Wistar Rats

This study addresses the possible protective effects of thymoquinone (TQ) against the development of experimentally‐induced benign prostatic hyperplasia (BPH) in Wistar rats. Eighteen adult male rats were divided into three groups; the negative control group (n = 6) received vehicle, and two groups received subcutaneous testosterone injection (3 mg/kg). Animals receiving testosterone were randomized to untreated BPH group (n = 6) and BPH + TQ treated group (n = 6, 50 mg/kg orally for 14 days). Histological changes and the mRNA levels of transforming growth factor‐β₁ (TGF‐β₁) and vascular endothelial growth factor‐A (VEGF‐A) were analyzed. Additionally, dihydrotestosterone and interleukin‐6 (IL‐6) serum levels were determined. The presented research shows significant increases in prostate weight/body weight ratio, prostate epithelial thickness, serum IL‐6 and dihydrotestosterone levels, and the prostatic expressions of TGF‐β₁ and VEGF‐A in the untreated BPH rats. Histological examination of the prostate tissues in the BPH rats showed an elevated level of proliferation in the stromal area and glandular epithelia with abundant intraluminal papillary folds. However, a reduction in prostate weight/body weight ratio, epithelial hyperplasia, serum IL‐6 levels, and the expressions of TGF‐β₁ and VEGF‐A were observed in the BPH + TQ treated rats compared with the untreated BPH rats. The findings support TQ as a useful natural treatment for animal BPH model. Copyright © 2017 John Wiley & Sons, Ltd.