Apoptosis induced by DICO, a novel non-aromatic B-ring flavonoid via a ROS-dependent mechanism in human colon cancer cells.

5,7-Dihydroxy-2-(1,2-isopropyldioxy-4-oxo-cyclohex-5-enyl)-chromen-4-one (DICO) is a novel non-aromatic B-ring flavonoid, isolated mainly from Macrothelypteris viridifrons and has anti-tumour properties. In this study, we investigated the cytotoxicity and underlying biochemical pathways leading to cell death, in response to DICO treatment of a human colon cancer cell line HT-29. Our results indicated that DICO induced apoptosis by elevating the generation of reactive oxygen species, which could be quenched by the antioxidants N-acetyl cysteine. In addition, activation of signal transducer and activator of transcription 3 and suppression of nuclear factor kappa B played a crucial role in DICO-induced apoptosis. Overall, our results provide mechanistic insights into the apoptotic action of a potential anti-tumour drug, DICO.

Inhibitory effect of diosgenin on experimentally induced benign prostatic hyperplasia in rats.

This study investigated the effect of diosgenin, a natural sapogenin possessing various pharmacological activities, on benign prostatic hyperplasia (BPH) in rats and the possible mechanisms. BPH was established in the castrated rats by subcutaneous injection of testosterone propionate. Animals were randomly divided into four groups (n=10 each): model group (0.5% sodium carboxymethyl cellulose); positive control group (3 mg/kg finasteride); two diosgenin groups (50 and 100 mg/kg). The drugs were intragastricaly given in each group for consecutive 3 weeks. Another 10 rats with no testicles cut off served as negative controls and they were subcutaneously injected with 0.1 mL olive oil per day and then treated with 0.5% sodium carboxymethylcellulose. After 3-week administration, the prostate index and serum PSA level were determined, and histopathological examination was carried out. The levels of MDA, SOD and GPx in prostates were also measured. Additionally, the expression of Bcl-2, Bax and p53 was examined using Western blotting. The results showed that the prostate index and serum PSA level were significantly decreased, and the pathological changes of the prostate gland were greatly improved in diosgenin groups as compared with the model group. Elevated activities of SOD and GPx, and reduced MDA level were also observed in diosgenin-treated rats. In addition, the expression of Bcl-2 in prostates was down-regulated, whereas that of Bax and p53 was up-regulated in diosgenin-treated rats. These results indicated that diosgenin was effective in inhibiting testosterone propionate-induced prostate enlargement and may be a candidate agent for the treatment of BPH.

Protective potential of the methanol extract of Macrothelypteris oligophlebia rhizomes for chronic non-bacterial prostatitis in rats.

The protective potential of the methanol extract of Macrothelypteris oligophlebia rhizomes (MMO) for chronic non-bacterial prostatitis (CNP) in rats was investigated in the present study. Carrageenan-induced CNP in rats was established. Fifty rats were randomly divided into sham-operated (sham-ope) group, model group, positive control group (Cernilton at a dose of 148mg/kg body weight) and two MMO-treated groups (MMO at doses of 600mg/kg and 300 mg/kg body weight). The anti-prostatitis effect was evaluated by prostate index, the levels of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-α), cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2), and histopathological examination. After 20 days of administration, MMO could significantly decrease prostate index and the levels of IL-10, TNF-α COX-2 and PGE2 in serum and could improve the prostate morphology in comparison with the model group. In summary, these results suggest that MMO possesses protective effects on prostate, which might be beneficial to further development for the treatment of CNP.

Neuroprotective effect of peptides extracted from walnut (Juglans Sigilata Dode) proteins on Aß25-35-induced memory impairment in mice.

Alzheimer's disease (AD) is one of the major neurodegenerative disorders of the elderly, which is characterized by the accumulation and deposition of amyloid-beta (Aß) peptide in human brains. Oxidative stress and neuroinflammation induced by Aß in brain are increasingly considered to be responsible for the pathogenesis of AD. The present study aimed to determine the protective effects of walnut peptides against the neurotoxicity induced by Aß25-35 in vivo. Briefly, the AD model was induced by injecting Aß25-35 into bilateral hippocampi of mice. The animals were treated with distilled water or walnut peptides (200, 400 and 800 mg/kg, p.o.) for five consecutive weeks. Spatial learning and memory abilities of mice were investigated by Morris water maze test and step-down avoidance test. To further explore the underlying mechanisms of the neuroprotectivity of walnut peptides, the activities of superoxide dismutase (SOD), glutathione (GSH), acetylcholine esterase (AChE), and the content of malondialdehyde (MDA) as well as the level of nitric oxide (NO) in the hippocampus of mice were measured by spectrophotometric method. In addition, the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), tumor necrosis factor-α (TNF-α), interleukin 1ß (IL-1ß) and IL-6 in the samples were determined using ELISA. The hippocampal expressions of inducible nitric oxide synthase (iNOS) and nuclear factor κB (NF-κB) were evaluated by Western blot analysis. The results showed that walnut peptides supplementation effectively ameliorated the cognitive deficits and memory impairment of mice. Meanwhile, our study also revealed effective restoration of levels of antioxidant enzymes as well as inflammatory mediators with supplementation of walnut peptides (400 or 800 mg/kg). All the above findings suggested that walnut peptides may have a protective effect on AD by reducing inflammatory responses and modulating antioxidant system.

Comparison of two thin-film microextractions for the analysis of estrogens in aqueous tea extract and environmental water samples by high performance liquid chromatography-ultraviolet detection.

Two thin-film microextractions (TFME), octadecylsilane (ODS)-polyacrylonitrile (PAN)-TFME and polar enhanced phase (PEP)-PAN-TFME have been proposed for the analysis of bisphenol-A, diethylstilbestrol and 17ß-estradiol in aqueous tea extract and environmental water samples followed by high performance liquid chromatography-ultraviolet detection. Both thin-films were prepared by spraying. The influencing factors including pH, extraction time, desorption solvent, desorption volume, desorption time, ion strength and reusability were investigated. Under the optimal conditions, the two TFME methods are similar in terms of the analytical performance evaluated by standard addition method. The limits of detection for three estrogens in environmental water and aqueous tea extract matrix ranged from 1.3 to 1.6 and 2.8 to 7.1 ng mL(-1) by the two TFME methods, respectively. Both approaches were applied for the analysis of analytes in real aqueous tea extract and environmental water samples, presenting satisfactory recoveries ranged from 87.3% to 109.4% for the spiked samples.

Preparation of magnetic molecularly imprinted polymers for selective isolation and determination of kaempferol and protoapigenone in Macrothelypteris torresiana.

Novel uniform-sized magnetic molecularly imprinted polymers (MMIPs) were synthesized for selective recognition of active antitumor ingredients of kaempferol (KMF) and protoapigenone (PA) in Macrothelypteris torresiana (M. torresiana) by surface molecular imprinting technique in this study. Super paramagnetic core-shell nanoparticles (γ-MPS-SiO2@Fe3O4) were used as seeds, KMF as template molecule, acrylamide (AM) as functional monomer, and N, N'-methylene bisacrylamide (BisAM) as cross-linker. The prepared MMIPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectrum (FTIR), transmission electron microscopy (TEM) and thermo-gravimetric analysis (TGA), respectively. The recognition capacity of MMIPs was 2.436 times of non-imprinted polymers. The adsorption results based on kinetics and isotherm analysis were in accordance with the pseudo-second-order model (R (2)=0.9980) and the Langmuir adsorption model (R (2)=0.9944). The value of E (6.742 kJ/mol) calculated from the Dubinin-Radushkevich isotherm model suggested that the physical adsorption via hydrogen-bonding might be predominant. The Scatchard plot showed a single line (R (2)=0.9172) and demonstrated the homogeneous recognition sites on MMIPs for KMF. The magnetic solid phase extraction (MSPE) based on MMIPs as sorbent was established for fast and selective enrichment of KMF and its structural analogue PA from the crude extract of M. torresiana and then KMF and PA were detected by HPLC-UV. The established method showed good performance and satisfactory results for real sample analysis. It also showed the feasibility of MMIPs for selective recognition of active structural analogues from complex herbal extracts.

Pallidifloside D, a saponin glycoside constituent from Smilax riparia, resist to hyperuricemia based on URAT1 and GLUT9 in hyperuricemic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots and rhizomes of Smilax riparia (SR), called "Niu-Wei-Cai" in traditional Chinese medicine (TCM), are believed to be effective in treating hyperuricemia and gout symptoms. This study was designed to isolate a saponin glycoside named pallidifloside D from the total saponins of Smilax riparia and to examine its effect in reducing serum uric acid levels in a hyperuricemic mouse model induced by potassium oxonate. MATERIALS AND METHODS: We examined the effects of pallidifloside D treated with 5, 10 and 20mg/kg on serum uric acid levels (SUA), Serum creatinine (SCr) and blood urea nitrogen (BUN) levels in a hyperuricemic mouse. A colorimetric method was used to evaluate the effects of pallidifloside D on the XOD activities, and Western Blotting analysis were carried out to observe protein levels of mURAT1, mGLUT9 and mOTA1 in hyperuricemic mice after treatment with pallidifloside D. RESULTS: The levels of serum uric acid levels (SUA) were suppressed significantly with dose-dependence by pallidifloside D treated with 5, 10 and 20mg/kg (p<0.05, p<0.01 and p<0.01 respectively). Pallidifloside D could down-regulate the expression levels of renal mURAT1 protein in hyperuricemic mice in a dose-dependent manner (p<0.05, p<0.01, and p<0.001 respectively), and the protein levels of mGLUT9 could be down-regulated with dose-dependence (p<0.05 and p<0.01 respectively) by pallidifloside D at the dose of 10 and 20mg/kg. CONCLUSION: These results suggest that pallidifloside D possesses a potent uricosuric effect in hyperuricemic mice through decreasing renal mURAT1 and GLUT9, which contribute to the enhancement of uric acid excretion and attenuate hyperuricemia-induced renal dysfunction.

Dioscin-induced apoptosis of human LNCaP prostate carcinoma cells through activation of caspase-3 and modulation of Bcl-2 protein family.

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin (1, 2 and 4 µmol/L) could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.

Neuroprotective effect of a caffeic acid derivative from Abacopteris penangiana.

CONTEXT: A new caffeic acid derivative, named (7'Z)-3-O-(3, 4-dihydroxyphenylethenyl)-caffeic acid (CADP), extracted from Abacopteris penangiana (Hook.) Ching. OBJECTIVE: To elucidate the neuroprotective effect of CADP against H2O2-induced cytotoxicity in PC12 cells and D-galactose (D-gal)-induced neurotoxicity in mice brain. MATERIALS AND METHODS: CADP was isolated from the methanol extract of the rhizomes of A. penangiana. In vitro, the protective effect of CADP (0.1-10 µM) against H2O2-induced oxidative damage on PC12 cells was investigated by a MTT assay. In vivo, behavioral tests and antioxidant enzymes measurements were performed to investigate the protective effect of intraperitoneal (i.p.) injection of CADP (5 or 10 mg/kg/day) for 2 weeks on D-gal-induced neurotoxicity in mice. RESULTS: The results showed that CADP significantly attenuated cell toxicity in a dose-dependent manner, and the EC50 value of CADP was 0.83 ± 0.02 µM. In vivo, it was found that CADP significantly improved the behavioral performance of D-gal-treated mice in both Morris water maze (MWM) test and step-down avoidance test. As compared with model group, CADP (5, 10 mg/kg/day) attenuated the decrease in superoxide dismutase (SOD) activities by 40.5 and 75.4%, respectively; attenuated the decrease in glutathione peroxidase (GPx) activities by 53.8 and 73.2%, respectively; attenuated the decrease in catalase activities by 12.0 and 53.3%, respectively; reduced the increased levels of malondialdehyde (MDA) by 38.6 and 79.9%, respectively. DISCUSSION AND CONCLUSION: The results suggested that CADP has significant neuroprotective effects which can be attributed to inhibiting the generation of free radical and enhancing the activity of endogenous antioxidant enzymes.

Fraction of macroporous resin from Smilax china L. inhibits testosterone propionate-induced prostatic hyperplasia in castrated rats.

The present study was conducted to evaluate the effect of a fraction of macroporous resin (FMR), a bioactive component of Smilax china L., on benign prostatic hyperplasia (BPH) in castrated rats induced by testosterone propionate. Rats were randomly divided into five groups: the negative control group (sham-operated), the model group, two FMR-treated groups (at doses of 300 mg/kg and 600 mg/kg of body weight), and the positive control group (treated with finasteride at the dose of 3 mg/kg). Drugs were administered once a day for three consecutive weeks by gastric gavage. Prostates were weighed, testosterone and dihydrotestosterone (DHT) levels in serum were determined, and histopathological examinations were carried out. FMR treatment inhibited prostatic hyperplasia, reducing the DHT level in serum and improving the prostate gland morphology compared with the model group. The overall results of this study suggest that FMR is effective at inhibiting experimentally induced prostate enlargement, and it presents a valuable resource for the treatment of human BPH.

Characterisation of phenols and antioxidant and hypolipidaemic activities of Lethariella cladonioides.

BACKGROUND: This study aimed to investigate the antioxidant and hypolipidaemic activities of an ethanol extract of Lethariella cladonioides (Nyl.) Krog (EE) and to characterise its chemical constituents. RESULTS: Nine phenols were identified as canarione, thamnolic acid, squamatic acid, vermicularin, norstictic acid, baeomycesis acid, lecanoric acid, barbatinic acid and usnic acid from analysis of EE by using high-performance liquid chromatography with a diode array detector-mass spectrometry. In antioxidant analysis in vitro, the highest scavenging rate of EEs on the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, superoxide anion, hydroxyl radicals and hydrogen peroxide was 81.55 ± 1.95%, 81.84 ± 4.00%, 74.28 ± 3.71% and 74.28 ± 3.71%, respectively. Meanwhile, after administration of EE for 6 weeks in high fat/cholesterol diet mice, the most significant reduction in levels of serum triglyceride, serum total cholesterol, serum low density lipoprotein cholesterol and liver malondialdehyde were 24%, 20%, 15% and 35%, respectively. The most significant increase in levels of serum high density lipoprotein cholesterol and liver superoxide dismutase was 35% and 88%, respectively. CONCLUSION: L. cladonioides possesses strong antioxidant and hypolipidaemic activities.

Hypolipidemic and anti-inflammatory properties of Abacopterin A from Abacopteris penangiana in high-fat diet-induced hyperlipidemia mice.

This study was to investigate the hypolipidemic and anti-inflammatory properties of Abacopterin A (APA), a flavonoid compound isolated from Abacopteris penangiana (Hook.) Ching. Male C57BL/6J mice were divided randomly and equally into five groups: the normal control group (N), the model group (M), the positive control group (P), the high and low doses of APA treated groups (H and L). All the animals except that in N group were fed with high-fat diet for 8 weeks. In the last 4 weeks, the mice in P, H and L groups were orally administered with simvastatin (at the dose of 20mg/kg/day) and APA (at the dose of 40 or 20mg/kg/day), respectively. Then the lipid profiles and related biochemical criterions of the studied mice were determined. The effects of high-fat diet on activating nuclear transcription factor-κB (NFκB) expression, elevating inflammatory factors tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) levels, and increasing triacylglycerol (TG) and total cholesterol (TC) levels were abolished on daily supplementation with APA. APA also enhanced lipoprotein lipase (LPL) and hepatic lipase (HL) activities. These results suggested that APA had hypolipidemic and anti-inflammatory properties through inhibiting NFκB expression, and reducing inflammatory response.

Neuropective constituents from the rhizomes of Abacopteris penangiana.

Two neuropective compounds were isolated from the rhizomes of Abacopteris penangiana, one was a new flavone and the other was a flavanone. Both compounds were firstly separated from natural plant. The isolation work was guided by the antioxidant activity. Both the compounds showed a significant antioxidant activity in vitro and a protective effect on dopamine-induced neurotoxicity in PC12 cells.

Two new acetylated flavan glycosides from rhizomes of the fern Abacopteris penangiana.

Two new acetylated flavan glycosides (2S,4R)-4,5,7-trihydroxy-4'-methoxy-6,8-dimethylflavan-5-O-ß-D-6-acetylglucopyranoside-7-O-ß-D-glucopyranoside (1) and (2S,4R)-5,7-dihydroxy-4,4'-dimethoxy-6,8-dimethylflavan-5-O-ß-D-6-acetylglucopyranoside-7-O-ß-D-glucopyranoside (2) were isolated from the rhizomes of Abacopteris penangiana. Their structures were elucidated on the basis of spectroscopic methods including IR, HR-ESI-MS, 1D and 2D NMR, and chemical evidences.

Design, synthesis, and cytoprotective effect of 2-aminothiazole analogues as potent poly(ADP-ribose) polymerase-1 inhibitors.

A series of novel poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors were designed within 2-aminothiazole analogues (4-10) based on a constructed three-dimensional pharmacophore model. After synthesis, the inhibitory effect on PARP-1 activity and the cytoprotective action of these compounds were tested and evaluated. Among them, compounds 4-6 and 10 appeared to be potent PARP-1 inhibitors with IC(50) values less than 1 microM, which had been perfectly predicted by pharmacophore model. These compounds proved to be highly potent against cell injury induced by H(2)O(2) and oxygen-glucose deprivation (OGD) in PC12 cells. These novel 2-aminothiazole analogues are potentially applicable as neuroprotective agents for the treatment of neurological diseases.

Three new polyphenols from the rhizomes of Arachniodes exilis.

Three new polyphenols, araspidin BB (1), arachniodesin A (2) and arachniodesin B (3), together with two known compounds, epicatechin (4) and procyanidin B-2 (5), were isolated from the rhizomes of Arachniodes exilis. The structures of three new compounds were elucidated as 5-methyl-methylene-bis-phlorobutyrophenone (1), 4beta-ethoxycarbonylmethylepicatechin (2) and epicatechin-(4beta --> 8)-4beta-ethoxycarbonylmethylepicatechin (3), on the basis of their spectral analysis and by comparing them with the related model compounds. Compounds 4 and 5 were obtained from the title plant for the first time.