Decursin in Angelica gigas Nakai (AGN) Enhances Doxorubicin Chemosensitivity in NCI/ADR‐RES Ovarian Cancer Cells via Inhibition of P‐glycoprotein Expression

Angelica gigas Nakai (AGN, Korean Dang‐gui) is traditionally used for the treatment of various diseases including cancer. Here, we investigated multidrug‐resistant phenotype‐reversal activities of AGN and its compounds (decursin, ferulic acid, and nodakenin) in doxorubicin‐resistant NCI/ADR‐RES ovarian cancer cells. Our results showed that a combination of doxorubicin with either AGN or decursin inhibited a proliferation of NCI/ADR‐RES cells. These combinations increased the number of cells at sub‐G1 phase when cells were stained with Annexin V‐fluorescein isothiocyanate. We also found that these combinations activated caspase‐9, caspase‐8, and caspase‐3 and increased cleaved PARP level. Moreover, an inhibition of P‐glycoprotein expression by either AGN or decursin resulted in a reduction of its activity in NCI/ADR‐RES cells. Therefore, our data demonstrate that decursin in AGN inhibits doxorubicin‐resistant ovarian cancer cell proliferation and induces apoptosis in the presence of doxorubicin via blocking P‐glycoprotein expression. Therefore, AGN would be a potentially novel treatment option for multidrug‐resistant tumors by sensitizing to anticancer agents. Copyright © 2016 John Wiley & Sons, Ltd.     

Auraptene Induces Apoptosis via Myeloid Cell Leukemia 1‐Mediated Activation of Caspases in PC3 and DU145 Prostate Cancer Cells

Although auraptene, a prenyloxy coumarin from Citrus species, was known to have anti‐oxidant, anti‐bacterial, antiinflammatory, and anti‐tumor activities, the underlying anti‐tumor mechanism of auraptene in prostate cancers is not fully understood to date. Thus, in the present study, we have investigated the anti‐tumor mechanism of auraptene mainly in PC3 and DU145 prostate cancer cells, because auraptene suppressed the viability of androgen‐independent PC3 and DU145 prostate cancer cells better than androgen‐sensitive LNCaP cells. Also, auraptene notably increased sub‐G1 cell population and terminal deoxynucleotidyl transferase dUTP nick end labeling‐positive cells as features of apoptosis in two prostate cancer cells compared with untreated control. Consistently, auraptene cleaved poly(ADP‐ribose) polymerase, activated caspase‐9 and caspase‐3, suppressed the expression of anti‐apoptotic proteins, including Bcl‐2 and myeloid cell leukemia 1 (Mcl‐1), and also activated pro‐apoptotic protein Bax in both prostate cancer cells. However, Mcl‐1 overexpression reversed the apoptotic effect of auraptene to increase sub‐G1 population and induce caspase‐9/3 in both prostate cancer cells. Taken together, the results support scientific evidences that auraptene induces apoptosis in PC3 and DU145 prostate cancer cells via Mcl‐1‐mediated activation of caspases as a potent chemopreventive agent for prostate cancer prevention and treatment. Copyright © 2017 John Wiley & Sons, Ltd.     

3‐O‐Acetyloleanolic Acid Induces Apoptosis in Human Colon Carcinoma Hct‐116 Cells

The cytotoxic effect of 3‐O‐acetyloleanolic acid, an oleanolic acid derivative isolated from the seeds of Vigna sinensis K., was investigated in human colon carcinoma HCT‐116 cells. 3‐O‐acetyloleanolic acid dose‐dependently inhibited the viability of HCT‐116 cells. Apoptosis was characterized by detection of cell surface annexin V and sub‐G1 apoptotic cell populations. The number of immunostained cells with annexin V‐FITC was increased after treatment with 3‐O‐acetyloleanolic acid. The sub‐G1 cell population was also increased. Expression of TRAIL‐mediated apoptosis signaling‐related death receptor DR5 was increased in 3‐O‐acetyloleanolic acid‐treated HCT‐116 cells. Activation of caspase‐8 and caspase‐3, critical mediators of extrinsic apoptosis signaling, was also increased by 3‐O‐acetyloleanolic acid. The results indicate that 3‐O‐acetyloleanolic acid induces apoptosis in HCT‐116 cells mediated by an extrinsic apoptosis signaling cascade via up‐regulation of DR5. Copyright © 2012 John Wiley & Sons, Ltd.     

Highly Oxygenated Sesquiterpene Lactones from Cousinia aitchisonii and their Cytotoxic Properties: Rhaserolide Induces Apoptosis in Human T Lymphocyte (Jurkat) Cells via the Activation of c‐Jun n‐terminal Kinase Phosphorylation

Infrared‐guided chromatographic fractionation of sesquiterpene lactones from the extracts of Cousinia aitchisonii and Cousinia concolor led to the isolation of five pure compounds. A new sesquiterpene lactone, namely, aitchisonolide, and two known sesquiterpene lactones (desoxyjanerin and rhaserolide) were isolated from C. aitchisonii and two known lignans (arctiin and arctigenin) from C. concolor. The structures of these compounds were elucidated by one‐dimensional and two‐dimensional nuclear magnetic resonance techniques, as well as high‐resolution mass spectrometry. The purified and characterized compounds were subjected to cytotoxicity assay. The sesquiterpene lactones desoxyjanerin and rhaserolide showed significant cytotoxic activities against five different cancer cell lines and the normal human embryonic kidney cell line. Rhaserolide was chosen to evaluate the possible mechanism of action. Western blot analysis revealed that rhaserolide could induce apoptosis in Jurkat cells via the activation of c‐Jun n ‐terminal kinase phosphorylation. Copyright © 2015 John Wiley & Sons, Ltd.     

Reactive Oxygen Species‐Mediated Activation of AMP‐Activated Protein Kinase and c‐Jun N‐terminal Kinase Plays a Critical Role in Beta‐Sitosterol‐Induced Apoptosis in Multiple Myeloma U266 cells

Although beta‐sitosterol has been well known to have anti‐tumor activity in liver, lung, colon, stomach, breast and prostate cancers via cell cycle arrest and apoptosis induction, the underlying mechanism of anti‐cancer effect of beta‐sitosterol in multiple myeloma cells was never elucidated until now. Thus, in the present study, the role of reactive oxygen species (ROS) in association with AMP‐activated protein kinase (AMPK) and c‐Jun N‐terminal kinase (JNK) pathways was demonstrated in beta‐sitosterol‐treated multiple myeloma U266 cells. Beta‐sitosterol exerted cytotoxicity, increased sub‐G₁ apoptotic population and activated caspase‐9 and ‐3, cleaved poly (ADP‐ribose) polymerase (PARP) followed by decrease in mitochondrial potential in U266 cells. Beta‐sitosterol promoted ROS production, activated AMPK, acetyl‐CoA carboxylase (ACC) and JNK in U266 cells. Also, beta‐sitosterol attenuated the phosphorylation of AKT, mammalian target of rapamycin and S6K, and the expression of cyclooxygenase‐2 and VEGF in U266 cells. Conversely, AMPK inhibitor compound C and JNK inhibitor SP600125 suppressed apoptosis induced by beta‐sitosterol in U266 cells. Furthermore, ROS scavenger N‐acetyl L‐cysteine attenuated beta‐sitosterol‐mediated sub‐G₁ accumulation, PARP cleavage, JNK and AMPK activation in U266 cells. Overall, these findings for the first time suggest that ROS‐mediated activation of cancer metabolism‐related genes such as AMPK and JNK plays an important role in beta‐sitosterol‐induced apoptosis in U266 multiple myeloma cells. Copyright © 2013 John Wiley & Sons, Ltd.     

Apoptotic Effect of Astragalin in Melanoma Skin Cancers via Activation of Caspases and Inhibition of Sry‐related HMg‐Box Gene 10

Though Astragalin (kaempferol‐3‐glucoside) contained in Paeonia lactiflora and other plants was known to have anti‐oxidant, antiinflammatory, and anti‐tumor activity, the anti‐tumor mechanism of Astragalin has never been reported in melanomas until now. Thus, in the present study, the underlying apoptotic mechanism of Astragalin isolated from Aceriphyllum rossii was elucidated in A375P and SK‐MEL‐2 melanoma cells. Astragalin exerted cytotoxicity in A375P and SK‐MEL‐2 cells in a concentration‐dependent manner. Also, Astragalin significantly increased the number of TdT‐mediated dUTP nick end labeling positive cells and sub‐G1 population as a feature of apoptosis in A375P and SK‐MEL‐2 cells compared with untreated control. Consistently, western blotting revealed that Astragalin activated caspase 9/3 and Bax, cleaved poly (ADP‐ribose) polymerase, and attenuated the expression of cyclin D1, Mcl‐1, and Sry‐related HMg‐Box gene 10 (SOX10) in A375P and SK‐MEL‐2 cells. Of note, ectopic expression of SOX10 reduced the apoptotic ability of Astragalin to inhibit proliferation, cleave poly (ADP‐ribose) polymerase, and caspase 3 in A375P and SK‐MEL‐2 melanoma cells. Overall, our findings provide evidence that Astragalin induces apoptosis in A375P and SK‐MEL‐2 melanoma cells via activation of caspase9/3 and inhibition of SOX10 signaling. Copyright © 2017 John Wiley & Sons, Ltd.     

Corosolic Acid Exhibits Anti‐angiogenic and Anti‐lymphangiogenic Effects on In Vitro Endothelial Cells and on an In Vivo CT‐26 Colon Carcinoma Animal Model

We describe the anti‐angiogenic and anti‐lymphangiogenic effects of corosolic acid, a pentacyclic triterpenoid isolated from Cornus kousa Burg. A mouse colon carcinoma CT‐26 animal model was employed to determine the in vivo anti‐angiogenic and anti‐lymphangiogenic effects of corosolic acid. Corosolic acid induced apoptosis in CT‐26 cells, mediated by the activation of caspase‐3. In addition, it reduced the final tumor volume and the blood and lymphatic vessel densities of tumors, indicating that it suppresses in vivo angiogenesis and lymphangiogenesis. Corosolic acid inhibited the proliferation and tube formation of human umbilical vein endothelial cells and human dermal lymphatic microvascular endothelial cells. In addition, corosolic acid decreased the proliferation and migration of human umbilical vein endothelial cells stimulated by angiopoietin‐1. Pretreatment with corosolic acid decreased the phosphorylation of focal adhesion kinase (FAK) and ERK1/2, suggesting that corosolic acid contains anti‐angiogenic activity that can suppress FAK signaling induced by angiopoietin‐1. Copyright © 2015 John Wiley & Sons, Ltd.     

[6]‐Shogaol Attenuates Neuronal Apoptosis in Hydrogen Peroxide‐Treated Astrocytes Through the Up‐Regulation of Neurotrophic Factors

Neuronal apoptosis induced by oxidative stress is a prominent feature of neurodegenerative disorders. [6]‐shogaol, a bio‐active compound in ginger, possesses potent anti‐inflammatory actions and has recently emerged as a potential therapeutic agent for neurodegenerative disorders. However, the effects of [6]‐shogaol on astroglial apoptosis following exogenously induced oxidative stress has not yet been investigated. Here, we show that the anti‐apoptotic activity of [6]‐shogaol in astrocytes following exposure to hydrogen peroxide (H₂O₂) involves a marked up‐regulation of neurotrophic factors such as nerve growth factor, glial cell line‐derived neurotrophic factor, and brain‐derived neurotrophic factor. Astrocytes co‐treated with [6]‐shogaol and H₂O₂ for 1 h showed decrease in reactive oxygen species production compared with those only treated with H₂O₂. Moreover, [6]‐shogaol counteracted the reduced expression of ERK1/2 in H₂O₂‐treated astrocytes and protected these cells from oxidative stress and apoptosis by attenuating the impairment of mitochondrial function proteins such as Bcl‐2 and Bcl‐xL. Additionally, [6]‐shogaol inhibits the expression of the apoptotic proteins Bax and caspase‐3 in H₂O₂‐treated astrocytes. This data suggest that following oxidative stress, [6]‐shogaol protects astrocytes from oxidative damage through the up‐regulating levels of neurotrophic factors. These findings provide further support for the use of [6]‐shogaol as a therapeutic agent in neurodegenerative disorders. Copyright © 2013 John Wiley & Sons, Ltd.     

Cytotoxic Impact of Costunolide Isolated from Costus speciosus on Breast Cancer via Differential Regulation of Cell Cycle—An In‐vitro and In‐silico Approach

Costunolide, a sesquiterpene lactone, is a biologically active molecule found in most of the medicinally valuable plants. The present study aims to evaluate the anticancer property of costunolide isolated from Costus speciosus against breast cancer cell lines (MCF‐7 and MDA‐MB‐231). Costunolide effectively reduced the viability of both MCF‐7 and MDA‐MB‐231 cell lines at an IC₅₀ value of 40 μM. Flow cytometric analysis revealed costunolide mediated cell cycle arrest at G2/M phase in both the cell types. Western blotting results confirmed the alterations in the expression of cell cycle regulators (cyclin D1, D3, CDK‐4, CDK‐6, p18 INK4c, p21 CIP1/Waf‐1 and p27 KIP1) and apoptosis inducers (caspase‐3 and caspase‐9) upon costunolide treatment in comparison with their expressions in normal breast cell line (MCF‐10A). Costunolide mediated downregulation of positive cell cycle regulators and upregulation of negative cell cycle regulators were related to the induction of apoptosis in cancer cells. The above results were validated with in‐silico results that predicted stable interactions between costunolide and cancer targets. Thus costunolide effectively induced breast cancer cell apoptosis targeting cell cycle regulation, and the compound can be used as an effective herbal therapeutic molecule to treat breast cancer with further explorations. Copyright © 2015 John Wiley & Sons, Ltd.     

Apoptosis Induction by 13‐Acetoxyrolandrolide through the Mitochondrial Intrinsic Pathway

The aim of this study was to evaluate the mechanisms of cytotoxicity of the sesquiterpene lactone 13‐acetoxyrolandrolide, a nuclear factor kappa B (NF‐κB) inhibitor that was previously isolated from Rolandra fruticosa. The effects associated with the inhibition of the NF‐κB pathway included dose‐dependent inhibition of the NF‐κB subunit p65 (RelA) and inhibition of upstream mediators IKKβ and oncogenic Kirsten rat sarcoma (K‐Ras). The inhibitory concentration of 13‐acetoxyrolandrolide on K‐Ras was 7.7 µm . The downstream effects of the inhibition of NF‐κB activation were also investigated in vitro. After 24 h of treatment with 13‐acetoxyrolandrolide, the mitochondrial transmembrane potential was depolarized in human colon cancer (HT‐29) cells. The mitochondrial oxidative phosphorylation was also negatively affected, and reduced levels of nicotinamine adenine dinucleotide phosphate (NAD(P)H) were detected after 2 h of 13‐acetoxyrolandrolide exposure. Furthermore, the expression of the pro‐apoptotic protein caspase‐3 increased in a concentration‐dependent manner. Cell flow cytometry showed that 13‐acetoxyrolandrolide induced cell cycle arrest at G₁, indicating that the treated cells had undergone caspase‐3‐mediated apoptosis, indicating negative effects on cancer cell proliferation. These results suggest that 13‐acetoxyrolandrolide inhibits NF‐κB and K‐Ras and promotes cell death mediated through the mitochondrial apoptotic pathway. Copyright © 2013 John Wiley & Sons, Ltd.     

β‐Bisabolene,a Sesquiterpene from the Essential Oil Extract of Opoponax (Commiphora guidottii), Exhibits Cytotoxicity in Breast Cancer Cell Lines

The essential oils from Commiphora species have for centuries been recognized to possess medicinal properties. Here, we performed gas chromatography‐mass spectrometry on the essential oil from opoponax (Commiphora guidotti) and identified bisabolene isomers as the main constituents of this essential oil. Opoponax essential oil, a chemical component; β‐bisabolene and an alcoholic analogue, α‐bisabolol, were tested for their ability to selectively kill breast cancer cells. Only β‐bisabolene, a sesquiterpene constituting 5% of the essential oil, exhibited selective cytotoxic activity for mouse cells (IC₅₀ in normal Eph4: >200 µg/ml, MG1361: 65.49 µg/ml, 4T1: 48.99 µg/ml) and human breast cancer cells (IC₅₀ in normal MCF‐10A: 114.3 µg/ml, MCF‐7: 66.91 µg/ml, MDA‐MB‐231: 98.39 µg/ml, SKBR3: 70.62 µg/ml and BT474: 74.3 µg/ml). This loss of viability was because of the induction of apoptosis as shown by Annexin V‐propidium iodide and caspase‐3/7 activity assay. β‐bisabolene was also effective in reducing the growth of transplanted 4T1 mammary tumours in vivo (37.5% reduction in volume by endpoint). In summary, we have identified an anti‐cancer agent from the essential oil of opoponax that exhibits specific cytotoxicity to both human and murine mammary tumour cells in vitro and in vivo, and this warrants further investigation into the use of β‐bisabolene in the treatment of breast cancers. Copyright © 2015 John Wiley & Sons, Ltd.     

Isolates from Alpinia officinarum Hance attenuate LPS‐induced inflammation in HepG2: Evidence from in silico and in vitro studies

In an attempt to connect the legacy of centuries of invaluable knowledge from traditional medicine and the current understanding to the molecular mechanism of diseases, we took the advantage of the emergence of in silico screening as a promising tool for identification of potential leads from libraries of natural products. Traditional Chinese Medicine database was subjected to structure based virtual screening for identification of anti‐inflammatory compounds using the 3D crystal structure of p38 alpha mitogen activated protein kinase. The molecular docking studies revealed the potential activity of several classes of compounds known to be the constituents of the rhizomes of Alpinia officinarum Hance (Lesser galangal). Five compounds, galangin, kaempferide, isorhamnetin, and two diarylheptanoids, were isolated from the rhizomes of the plant using vacuum liquid chromatography and flash chromatography techniques. The anti‐inflammatory activity of these compounds was investigated on HepG2 cells stimulated by lipopolysaccharide. The latter induced the gene expression of proinflammatory cytokines; interleukin‐1β, interleukin‐6, tumor necrosis factor alpha. Addition of the 5 isolated compounds downregulated this increased gene expression in a dose dependent manner. Thus, these results indicate that the isolated compounds from A. officinarum could be used as a beneficial source for preventing and treating inflammatory diseases.     

Phloroglucinol Protects INS‐1 Pancreatic β‐cells Against Glucotoxicity‐Induced Apoptosis

Decreasing numbers, and impaired function, of pancreatic β‐cells are key factors in the development of type 2 diabetes. This study was designed to investigate whether phloroglucinol protected pancreatic β‐cells against glucotoxicity‐induced apoptosis using a rat insulinoma cell line (INS‐1). High glucose treatment (30 mM) induced INS‐1 cell death; however, the level of glucose‐induced apoptosis was significantly reduced in cells treated with 100‐μM phloroglucinol. Treatment with 10–100‐μM phloroglucinol increased cell viability and decreased intracellular levels of reactive oxygen species, nitric oxide, and lipid peroxidation dose‐dependently in INS‐1 cells pretreated with high glucose. Furthermore, phloroglucinol treatment markedly reduced the protein expression of Bax, cytochrome c, and caspase 9, while increasing anti‐apoptotic Bcl‐2 protein expression. Cell death type was examined using annexin V/propidium iodide staining, revealing that phloroglucinol markedly reduced high glucose‐induced apoptosis. These results demonstrated that phloroglucinol could be useful as a potential therapeutic agent for the protection of pancreatic β‐cells against glucose‐induced apoptosis. Copyright © 2015 John Wiley & Sons, Ltd.     

Pachymic Acid Induces Apoptosis of EJ Bladder Cancer Cells by DR5 Up‐Regulation,ROS Generation,Modulation of Bcl‐2 and IAP Family Members

Pachymic acid (PA) is a lanostane‐type triterpenoid derived from Poria cocos mushroom that possess various biological effects such as anti‐cancer, antiinflammatory and anti‐metastasis effects. In this study, we investigated the anti‐cancer effects of PA in EJ bladder cancer cells. The results showed that PA significantly inhibited proliferation of EJ cells in a dose‐dependent manner. PA induced accumulation of sub‐G1 DNA content (apoptotic cell population), apoptotic bodies and chromatin condensation and DNA fragmentation in EJ cells in a dose‐dependent manner. PA also induces activation of caspase‐3, ‐8 and ‐9, and subsequent cleavage of poly (ADP‐ribose) polymerase, and significantly suppressed the inhibitor of apoptosis protein family proteins in a dose‐dependent manner. Furthermore, PA activates Bid and induced the loss of mitochondrial membrane potential (ΔΨ_m) with up‐regulated pro‐apoptotic proteins (Bax and Bad), down‐regulated anti‐apoptotic proteins (Bcl‐2 and Bcl‐xL) and cytochrome c release. In turn, PA increased the generation of reactive oxygen species (ROS); also, the ROS production was blocked by N‐acetyl‐L‐cysteine. The expressions of TNF‐related apoptosis inducing ligand and death receptor 5 were up‐regulated by PA in a dose‐dependent manner, suggesting extrinsic pathway also involved in PA‐induced apoptosis. This study provides evidence that PA might be useful in the treatment of human bladder cancer. Copyright © 2015 John Wiley & Sons, Ltd.     

Apoptotic Effect of Galbanic Acid via Activation of Caspases and Inhibition of Mcl‐1 in H460 Non‐Small Lung Carcinoma Cells

Galbanic acid (GBA), a major compound of Ferula assafoetida, was known to have cytotoxic, anti‐angiogenic and apoptotic effects in prostate cancer and murine Lewis lung cancer cells; the underling apoptotic mechanism of GBA still remains unclear so far. Thus, in the present study, the apoptotic mechanism of GBA was investigated mainly in H460 non‐small cell lung carcinoma (NSCLC) cells because H460 cells were most susceptible to GBA than A549, PC‐9 and HCC827 NSCLC cells. Galbanic acid showed cytotoxicity in wild EGFR type H460 and A549 cells better than other mutant type PC‐9 and HCC827 NSCLC cells. Also, GBA significantly increased the number of Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) positive cells and sub G1 population in H460 cells. Western blotting revealed that GBA cleaved poly (ADP‐ribose) polymerase (PARP), activated Bax and caspase 9, attenuated the expression of Bcl‐2, Bcl‐x_L, and Myeloid cell leukemia 1 (Mcl‐1) in H460 cells. However, interestingly, overexpression of Mcl‐1 blocked the ability of GBA to exert cytotoxicity, activate caspase9 and Bax, cleave PARP, and increase sub G1 accumulation in H460 cells. Overall, these findings suggest that GBA induces apoptosis in H460 cells via caspase activation and Mcl‐1 inhibition in H460 cells as a potent anticancer agent for NSCLC treatment. Copyright © 2015 John Wiley & Sons, Ltd.     

Antioxidant and Anti‐α‐glucosidase Compounds from the Rhizome of Peltiphyllum peltatum (Torr.) Engl

The antioxidant, anti‐α‐glucosidase and anticholinesterase effects of the alcohol extract of fresh underground rhizomes of Peltiphyllum peltatum were studied. A potent antioxidant activity accompanied by a selective α‐glucosidase effect was observed for the crude extract. Further activity‐guided fractionation (petroleum ether, chloroform, ethyl acetate, n‐butanol and water) resulted in the identification of the ethyl acetate fraction with the highest antioxidant effect. Gallic acid, methyl‐3‐O‐methyl gallate, catechin, gallocatechin, bergenin and 11‐O‐galloylbergenin were isolated from the ethyl acetate fraction. While all the isolated compounds did show a variable degree of radical scavenging effect, 11‐O‐galloylbergenin was identified as the selective α‐glucosidase inhibitor. The isolation, structural elucidation and biological effects of these compounds are discussed. Copyright © 2012 John Wiley & Sons, Ltd.     

Anti‐leishmanial Activities of Extracts and Isolated Compounds from Drechslera rostrata and Eurotium tonpholium

The fungal extract of Drechslera rostrata and Eurotium tonpholium showed a significant anti‐leishmanial activity against Leishmania major; IC₅₀ was 28.8 and 28.2 μg/mL, respectively. Seven compounds, five from D. rostrata (H1–H5) and two from E. tonpholium (H6 and H7), were isolated and identified using different spectroscopic analysis including ¹HNMR, ¹³CNMR, Hetero‐nuclear multiple bond connectivity (HMBC), Hetero‐nuclear Multiple Quantum Correlation (HMQC), and EI‐MS. The isolated compounds are: di‐2‐ethylhexyl phthalate (1), (22E)‐5α,8α‐epidioxyergosta‐6,22‐diene‐3β‐ol (2),1,3,8‐trihydroxy‐6‐methyl‐nthraquinone (3), aloe‐emodine 8‐O‐glucopyranoside(4), 2R, 3R,4R,5R hexane 1, 2, 3, 4, 5, 6 hexole (Mannitol) (5), 1,8‐dihydroxy‐3‐methoxy‐6‐methyl‐anthraquinone (6) and 1, 4, 5‐trihydroxy‐7‐methoxy‐2‐methyl‐anthraquinone (7). However, compounds (1) and (6) showed activity against L. major with IC₅₀ of 3.2 and 10.38 µg/mL, respectively. On the other hand, oral administration of the two extracts (100 mg/kg) and compounds 1 and 6 (50 mg/kg) showed very good activity when compared with the anti‐leishmanial drug Pentostam (125 mg/kg). Interestingly, the complete heeling activity of the extracts and compounds (1) and (6) was obtained after 13–17 days of treatment, while complete healing activity of Pentostam was obtained after 28 days. No alteration on liver and kidney functions was recorded on animals treated with the two extracts for 15 consecutive days. Copyright © 2013 John Wiley & Sons, Ltd.     

Anti‐inflammatory Activities of Eleven Centaurea Species Occurring in the Carpathian Basin

Our study aimed at the identification of anti‐inflammatory activities of different fractions of C. sadleriana extract after per os administration in rats, the identification of the active compounds of the plant and the investigation of the in vitro anti‐inflammatory activities of Centaurea species native to or cultivated in the Carpathian Basin. The aerial parts of Centaurea sadleriana Janka have been used in Hungarian folk medicine to treat the wounds of sheep. Methanol extract of C. sadleriana was fractioned by solvent‐solvent partitioning. The n‐hexane fraction was further fractionated and the anti‐inflammatory activities of certain subfractions were confirmed in vivo in rats. The n‐hexane and chloroform fraction of the methanol extract of C. sadleriana exhibited remarkable COX‐1 and COX‐2 inhibiting effects in vitro. Chromatographic separation of the fractions led to the identification of the active subfractions and 11 compounds (α‐linolenic acid, γ‐linolenic acid, stigmasterol, β‐sitosterol, campesterol, vanillin, pectolinarigenin, salvigenin, hispidulin, chrysoeriol and apigenin). The in vitro screening for anti‐inflammatory activities of further Centaurea species occurring in the Carpathian Basin (C. adjarica, C. bracteata, C. cataonica, C. cynaroides, C. dealbata, C. indurata, C. macrocephala, C. melitensis, C. nigrescens, C. ruthenica) revealed considerable COX‐1 and COX‐2 inhibitory activities. Because C. sadleriana is an endangered species native only to the Carpathian Basin, the investigation of more prevalent species is reasonable. Copyright © 2012 John Wiley & Sons, Ltd.