Jinlan Ruan

Abacopteris penangiana exerts testosterone-induced benign prostatic hyperplasia protective effect through regulating inflammatory responses, reducing oxidative stress and anti-proliferative.

ETHNOPHARMACOLOGICAL RELEVANCE Abacopteris penangiana (Hook.) Ching (AP) is a member of parathelypteris glanduligera and used in folk medicine for the treatment of blood circulation and blood stasis, edema and inflammation as recorded in the ″Chinese Materia Medica″. AIM OF THE STUDY The purpose of this study was to investigate the effects of total flavanol glycosides (TFA) from AP and its acid hydrolysate (AHT) on testosterone-induced benign prostatic hyperplasia (BPH) in rats by measuring the levels of inflammatory responses, oxidative stress and prostate cell proliferation. MATERIALS AND METHODS BPH was induced in rats by subcutaneous injection of testosterone after castration. Seventy rats were divided into seven groups. After oral administration of AHT and TFA (100 or 200mg/kg/d) for 4 weeks, the prostate index (PI), 5a-reductase (5α-R) and dihydrotestosterone (DHT) were determined. Then the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were determined. In addition, the relative inflammatory factors, cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8) and interleukin 17 (IL-17) were measured. Finally, the prostatic expression of nuclear transcription factor-κB (NF-κB) and phosphoinositide3-kinase (PI3K)/Akt were determined by immunohistochemistry. The prostatic expression of Bcl-2 was determined by western blot analysis. RESULTS The results showed that AHT and TFA decreased serum DHT and 5α-R activities compared with model group, as well as the PI and histopathological examination findings. In addition, oral treatment of AHT and TFA can significantly increase the activities of SOD, GPx and CAT while the level of MDA was significantly decreased compared with the model group. Moreover, AHT and TFA remarkably decreased the levels of inflammatory cytokines in prostatic tissue. Further investigation demonstrated that AHT and TFA treatment down-regulated the protein expressions of p-Akt, NF-κB and Bcl-2. CONCLUSIONS These results suggest that AHT and TFA have anti-BPH properties via anti-inflammatory, antioxidant and anti-proliferative effects. Hence, AP represents a potential herb for the treatment of BPH.

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

SummaryDioscin 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.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.

The furano norclerodane diterpenoid disobulbin-D induces apoptosis in normal human liver L-02 cells.

Disobulbin-D (DBD), a hepatotoxic furano norclerodane diterpenoid, was isolated by bio-guided fractionation from the rhizome of Dioscorea bulbifera L. In working toward elucidating the cellular and molecular mechanisms of DBD toxicity, we treated normal human liver cell line L-02 cells with DBD in vitro and evaluated its toxicity in terms of cell viability, morphologic changes, induction of apoptosis/necrosis, and caspase 3 activity. The viability of L-02 cells was inhibited by DBD in a concentration and time-dependent manner. Apoptosis was supported by the Annexin V and propidium iodide assay, Hoechst 33258 staining, and the occurrence of a sub-G(1) peak. DBD can cause an increase in caspase 3 activity, and pretreatment with Ac-DEVD-CHO blocked cell death and attenuated the apoptosis, showing that DBD-induced L-02 cell apoptosis is caspase 3-dependent. These results suggest that the effects of DBD on the growth of normal human liver L-02 cells may be due to its induction of cell apoptosis, which may also explain the toxicity observed in the plants containing furano clerodane diterpenoids.

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 17β-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.

Apoptosis Induction by the Total Flavonoids from Arachniodes exilis in HepG2 Cells through Reactive Oxygen Species-Mediated Mitochondrial Dysfunction Involving MAPK Activation

Arachniodes exilis is used as a folk medicine in China and proved to have antibacterial, anti-inflammatory, and sedative activities. In the present study, the antitumor effect of the total flavonoids of A. exilis (TFAE) against HepG2 cells was evaluated. The results showed that TFAE inhibited the growth of HepG2 cells in a dosage- and time-dependent manner. Flow cytometry and Hoechst 33342 fluorescence staining results showed that TFAE could significantly increase the apoptosis ratio of HepG2 cells, which is accompanied with increased intracellular reactive oxygen species (ROS) production and decreased mitochondrial membrane potential (ΔΨm). Western blotting indicated that TFAE downregulated the ratio of Bcl-2/Bax, increased cytochrome c release, and activated the caspases-3 and -9. Further analysis showed that TFAE stimulated the mitogen-activated protein kinase (MAPK). However, treatment with NAC (reactive oxygen species scavenger) and MAPK-specific inhibitors (SP600125 and SB203580) could reverse the changes of these apoptotic-related proteins. These results suggested that TFAE possessed potential anticancer activity in HepG2 cells through ROS-mediated mitochondrial dysfunction involving MAPK pathway.

Multitargeted protective effect of Abacopteris penangiana against carrageenan-induced chronic prostatitis in rats

ETHNOPHARMACOLOGICAL RELEVANCE Abacopteris penangiana (Hook.) Ching (AP) is traditionally used in Chinese medicine to promote blood circulation, remove blood stasis and dampness and for the treatment of edema and inflammation. In order to further support and develop the traditional use of Abacopteris penangiana as Chinese folk medicine, the aim of this study is to investigate the protective effect of the total flavanol glycosides (TFA) from AP and its acid hydrolysate (AHT) on chronic non-bacterial prostatitis (CNP) by measuring the levels of oxidative stress and inflammatory responses in rats. MATERIALS AND METHODS First, the antioxidant and anti-inflammatory activities of AHT and TFA were investigated. Then the experimental chronic non-bacterial prostatitis was induced by carrageenan. The prostate index (PI) and prostate specific antigen (PSA) were determined. The activities of AHT and TFA on inhibiting free radicals and oxidative stress were investigated. Subsequently, the degree of chronic inflammatory cell infiltrates, acinar changes and interstitial fibrosis were evaluated by histopathological examination. In addition, the relative inflammatory factors, tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), cyclooxygenase-2 (COX-2), prostaglandin E2 (PEG2), transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) were measured. Finally, the prostatic expression of nuclear transcription factor-κB (NF-κB) was determined by immunohistochemistry and western blot analysis. RESULTS The whole results showed that AHT and TFA had strong antioxidant and anti-inflammatory activities. In CNP model, AHT and TFA successfully decreased PI and PSA. The activities of antioxidant enzymes in AHT or TFA group were enhanced. Additionally, a morphometric analysis of the prostate gland of AHT or TFA treated rats demonstrated a significant reduction in chronic inflammatory cell infiltrates and interstitial fibrosis compared to model group. The reduced values of TNF-α, IL-1β, COX-2, PEG2, inducible nitric oxide synthase (iNOS) and nitric oxide (NO) were observed both in AHT and TFA treated groups. Moreover, the levels of TGF-β1 and CTGF in AHT and TFA treated groups were significantly decreased along with the alleviation of the inflammatory state of the prostate gland. Besides, the prostatic expression of NF-κB was inhibited. CONCLUSIONS These results suggest that AHT and TFA have anti-prostatitis properties via inhibiting oxidative stress, NF-κB dependent pro-inflammatory cytokines, fibrosis-related factors and antinociceptive activity. Hence, AP represents a potential herb for the treatment of prostatitis.

Neuroprotective effect of peptides extracted from walnut (Juglans Sigilata Dode) proteins on Aβ25-35-induced memory impairment in mice.

SummaryAlzheimer’s disease (AD) is one of the major neurodegenerative disorders of the elderly, which is characterized by the accumulation and deposition of amyloid-beta (Aβ) peptide in human brains. Oxidative streβs and neuroinflammation induced by Aβ 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 Aβ25-35 in vivo. Briefly, the AD model was induced by injecting Aβ25-35 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 1β (IL-1β) 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.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 (Aβ) peptide in human brains. Oxidative streβs and neuroinflammation induced by Aβ 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 Aβ25-35 in vivo. Briefly, the AD model was induced by injecting Aβ25-35 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 1β (IL-1β) 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.

Toxicity of a Diterpene Lactone Isolated from Dioscorea bulbifera on Hepatocytes

Abstract Aim To test the toxic effects of a diterpene lactone, diosbulbin-D, isolated from Dioscorea bulbifera L. on hepatocytes. Method Using the MTT assay to test the effect of diosbulbin-D on hepatocytes. Selected intracellular enzymes release levels by diosbulbin-D were determined to further validate the toxic effect on hepatocytes. Furthermore, the level of intracellular glutathione and the induction of reactive oxygen species (ROS) by diosbulbin-D were measured by fluorometric and flow cytometry methods. Results Diosbulbin-D showed toxic effect on hepatocytes and a significant increase in intracellular release levels of LDH and the liver enzymes, alanine aminotransferase and aspartate aminotransferase confirmed diosbulbin-Ds toxic effect. ROS increase was related to the decrease in level of GSH. Cell pretreated NAC almost completely blocked the ROS fluorescence and recovered the cell growth inhibition induced by diosbulbin-D. Conclusion Diosbulbin-D shows direct toxic effect on hepatocytes. The mechanism could be associated with oxidative stress.

Enrichment and purification of flavones from rhizomes of Abacopteris penangiana by macroporous resins

Abstract Aim To investigate the enrichment and purification of flavones from the rhizomes of Abacopteris penangiana (RAP) by macroporous resins. Methods Static adsorption and desorption tests were performed to select the appropriate resin. The kinetic adsorption and desorption experiments were carried out on selected HPD500 resin to optimize the separation process of flavones. Additionally, the effects of four parameters including adsorption flow rate, elute flow rate, volume of water and ethanol solution for elution were explored by a L4/3 orthogonal experiment. Finally, the ABTS (2, 2′-azinobis-3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activities of samples before and after being treated by HPD500 were compared. Results The results showed that the optimal parameters were initial concentration of 2.86 mgmL −1 , elute solution of 70% ethanol, absorb flow rate of 1 mLmin −1 , elute flow rate of 2 mLmin −1 , 5 BV of water for elution and 5 BV of ethanol solution for elution. Conclusion The content of flavones is above 60% in RAP after being treated by HPD500, indicating that macroporous resins could be successfully applied to enrich and purify flavones in RAP.

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.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.