Protective effects of aqueous extract from Acanthopanax senticosus against corticosterone-induced neurotoxicity in PC12 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Acanthopanax senticosus, classified into the family of Araliaceae, has been known for thousands of years as a remedy and is used to treat various diseases in traditional Chinese medicine system including hypertension, ischemic heart disease and hepatitis. AIM OF THE STUDY: This study aimed to examine the protective effects of aqueous extract from Acanthopanax senticosus (ASE) on corticosterone-induced neurotoxicity and its possible mechanisms, using PC12 cells as a suitable in vitro model of depression. MATERIALS AND METHODS: In this paper, PC12 cells were treated with 200 µM of corticosterone in the absence or presence of ASE in varying concentrations for 24 h. Then, cell viability was measured by MTT assay. The release amount of lactate dehydrogenase (LDH) was quantified using LDH assay kit. Apoptosis of PC12 cells was measured by Annexin V-FITC and PI labeling. The intracellular Ca(2+) content was tested by fluorescent labeling. The mRNA level of brain-derived neurotrophic factor (BDNF) was examined by real-time RT-PCR, and the expression of cAMP response element binding protein (CREB) was determined by western blotting. RESULTS: The results showed that treatment with 200 µM of corticosterone could induce cytotoxicity in PC12 cells. However, different concentrations of ASE (50, 100, 200, and 400 µg/mL) significantly increased the cell viability, decreased the LDH release, suppressed the apoptosis of PC12 cells, attenuated the intracellular Ca(2+) overloading, up-regulated the BDNF mRNA level and CREB protein expression compared with the corresponding corticosterone-treated group. CONCLUSION: The present results suggest that ASE exerts a neuroprotective effect on corticosterone-induced neurotoxicity in PC12 cells, which may be one of the acting mechanisms that accounts for the in vivo antidepressant activity of ASE.

Anti-depressant effects of aqueous extract from Acanthopanax senticosus in mice.

In this paper, the anti-depressant effects of Acanthopanax senticosus extract (ASE) were studied using animal models of depression including the forced swimming and tail suspension tests. The anti-depressive mechanism of ASE was explored by monitoring the levels of monoamine neurotransmitters including 5-hydroxytrylamine (5-HT), norepinephrine (NE), and dopamine (DA), as well as cAMP response element-binding (CREB) protein expression in the whole brain of mice following the tail suspension test. Our results showed that intragastric administration of ASE at a dose of 2000 mg/kg for seven days significantly reduced the duration of immobility in both the forced swimming test and the tail suspension test. These results indicate that ASE possesses antidepressant-like properties. Pre-treatment with 2000 mg/kg of ASE for seven days significantly elevated the levels of 5-HT, NE, and DA in the whole brain of mice. Moreover, ASE at doses of 1000 and 2000 mg/kg significantly up-regulated the level of CREB protein. Taken together, these findings suggest that the anti-depressive mechanism of ASE may be mediated via the central monoaminergic neurotransmitter system and CREB protein expression. Therefore, administration of ASE may be beneficial for patients with depressive disorders.

Deer antler base as a traditional Chinese medicine: a review of its traditional uses, chemistry and pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Deer antler base (Cervus, Lu Jiao Pan) has been recorded in the Chinese medical classics Shen Nong Ben Cao Jing 2000 years ago and is believed to nourish the Yin, tonify the kidney, invigorate the spleen, strengthen bones and muscles, and promote blood flow. In China, deer antler base has been extensively used in traditional Chinese medicine (TCM) to treat a variety of diseases including mammary hyperplasia, mastitis, uterine fibroids, malignant sores and children's mumps. AIM OF THE REVIEW: We provide an up-to-date and comprehensive overview of the traditional uses, chemistry, pharmacology, toxicology and clinical trials of deer antler base in order to explore its therapeutic potentials and future research needs. BACKGROUND AND METHODS: The pharmacological value of deer antler base was ignored for many years while researchers concentrated on the pharmacological value of velvet antler. However, more recently, scientists have carried out a great number of chemical, pharmacological and clinical studies on deer antler base. The present review covers the literature available from 1980 to 2012. All relevant information on deer antler base was collected from ancient Chinese herbal classics, pharmacopoeias, formularies, scientific journals, books, theses and reports via a library and electronic search by using PubMed, Google Scholar, Web of Science, Science Direct, and CNKI (in Chinese). KEY FINDINGS: Both in vitro and in vivo pharmacological studies have demonstrated that deer antler base possess immunomodulatory, anti-cancer, anti-fatigue, anti-osteoporosis, anti-inflammatory, analgesic, anti-bacterial, anti-viral, anti-stress, anti-oxidant, hypoglycemic, hematopoietic modulatory activities and the therapeutic effect on mammary hyperplasia. Although the mechanism of actions is still not clear, the pharmacological activities could be mainly attributed to the major bioactive compounds amino acids, polypeptides and proteins. Based on animal studies and clinical trials, deer antler base causes no severe side effects. CONCLUSIONS: Deer antler base has emerged as a good source of traditional medicine. However, further investigations are needed to explore individual bioactive compounds responsible for these in vitro and in vivo pharmacological effects and its mechanism of actions. Further safety assessments and clinical trials in humans need to be performed before it can be integrated into medicinal practices. The present review has provided preliminary information for further studies and commercial exploitations of deer antler base.

Protective effects of aucubin on H2O2-induced apoptosis in PC12 cells.

The present study investigated the neuroprotective effects of aucubin on hydrogen peroxide (H2O2)-induced apoptosis in PC12 cells. Exposure of PC12 cells to 0.25 mm H2O2 induced a leakage of lactate dehydrogenase and decreased cell viability, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In a dose over 0.1 mm, aucubin increased PC12 cellular viability and markedly attenuated H2O2-induced apoptotic cell death. Quantitation of apoptosis by flow cytometry indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells. Nuclear damage was alleviated by aucubin, as shown by Hoechst staining. In addition, the levels of malondialdehyde were reduced and the activity of superoxide dismutase, catalase and glutathione peroxidase was augmented in these cells. These results indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells through regulation of the endogenous oxidant-antioxidant balance. Our results suggest that aucubin is a potential protective agent for the treatment of oxidative-stress-induced neurodegenerative disease.

Aucubin prevents loss of hippocampal neurons and regulates antioxidative activity in diabetic encephalopathy rats.

In this study, the neuroprotection of aucubin and its mechanism were evaluated in the rat model of diabetic encephalopathy. Diabetes mellitus (DM) rats were stratified by cognitive capability (CC), and assigned to four treatment groups for aucubin treatment (doses of 0, 1, 5 or 10 mg/kg aucubin), with a further two groups of non-DM rats ranked by CC as controls for aucubin (doses of 0 or 5 mg/kg aucubin). Neuroprotection was estimated by the indexes of behavior and histology. Behavioral testing was performed in a Y-maze. The surviving neurons in CA1-CA4 and subiculum (SC) of the hippocampus were counted under a microscope. In addition, the apoptotic neurons in the CA1 of the hippocampus were also examined by using TUNEL staining. In order to clarify the mechanism of aucubin's neuroprotection, the activities of endogenous antioxidants and nitric oxide synthase (NOS) together with the content of lipid peroxide in the hippocampus were assayed. The results proved that aucubin significantly reduced the content of lipid peroxide, regulated the activities of antioxidant enzymatic and decreased the activity of NOS. All these effects indicated that aucubin was a potential neuroprotective agent and its neuroprotective effects were achieved, at least in part, by promoting endogenous antioxidant enzymatic activities.

Tanshinone IIA interacts with DNA by minor groove-binding.

Tanshen has long been widely used as a traditional Chinese medicine. Tanshinone IIA (Tan IIA) is the most abundant lipophilic constituent of Tanshen which has antitumor activity but the mechanism is poorly understood. Some preliminary reports hypothesized that it is a DNA intercalator and that the furano-o-quinone moiety could produce free radicals responsible for its cytotoxicity. Here the interaction of Tan IIA with DNA was explored in detail using fluorescence, viscosimetry, and molecular modeling. Tan IIA was found to bind with DNA in the minor groove rather than act as an intercalator. Furthermore, the results of immunofluorescence showed that Tan IIA does not produce free radicals in vivo to damage DNA. The former hypothesis was thus negated. The furan oxygen plays the key role in the antitumor ability of Tan IIA because it is involved in the groove-binding, but not in the production of free radicals. The molecular basis illustrated here could be responsible for all the findings in the structure-relationship studies of tanshinone cytotoxicity.

Protective effect of Acanthopanax senticosus extract against endotoxic shock in mice.

AIM OF THE STUDY: In this study, we evaluated protective effect of Acanthopanax senticosus extract (ASE) and a possible signaling pathway involved during endotoxic shock induced by intraperitoneal injection lipopolysaccharide (LPS) and D-galactosamine (D-GalN) in BALB/c mice. MATERIALS AND METHODS: Mice were intraperitoneal administrated with ASE (100, 200 or 400mg/kg) prior to injection of 50 microg/kg LPS and 1g/kg D-GalN. The levels of tumor necrosis-alpha (TNF-alpha) and interleukin-10 (IL-10) in serum and liver. Nitric oxide (NO) production in serum and inducible nitric oxide synthase (iNOS) protein level were investigated. Nuclear factor-kappa B (NF-kappaB) activation in liver was determined. Furthermore, we evaluated the effect of ASE pretreatment on infiltration of inflammatory cells into the heart, liver and lung of mice. RESULTS: Treatment of mice with ASE prior to LPS/D-GalN injection significantly improved the survival rate. ASE pretreatment inhibited the elevation of TNF-alpha in serum and liver. ASE also decreased iNOS level in liver and the overproduction of nitric oxide (NO) in serum. In addition, IL-10 levels in serum and liver were markedly enhanced. ASE pretreatment inhibited NF-kappaB activation in liver of mice. Moreover, infiltration of inflammatory cells into the heart, liver and lung of mice was also attenuated by ASE pretreatment. CONCLUSIONS: These results suggested that ASE protected mice against LPS/D-GalN-induced endotoxic shock involving inhibition of NF-kappaB activation, which caused down-regulation of TNF-alpha and involved up-regulation of IL-10. Acanthopanax senticosus may thus prove beneficial in the prevention of endotoxic shock.

Acanthopanax senticosus suppresses reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo.

Excess production of reactive oxygen species by macrophages has been implicated in many inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus on the production of superoxide anion and hydrogen peroxide in mouse peritoneal macrophages in vitro and in vivo. Exposure of mouse peritoneal macrophages to A. senticosus extract significantly suppressed superoxide anion production induced by zymosan in a dose-dependent manner. Similarly, exposure of mouse peritoneal macrophages to A. senticosus extract significantly inhibited hydrogen peroxide production induced by phorbol 12-myristate 13-acetate (PMA) in a dose-dependent manner. Intraperitoneal administration of A. senticosus extract to KM mice reduced the ex vivo production of zymosan induced-superoxide anion and PMA-induced hydrogen peroxide by their peritoneal macrophages. Exposure to A. senticosus extract did not affect the cell viability or systemic toxicity. A. senticosus inhibited reactive oxygen species production by mouse peritoneal macrophages in vitro and in vivo and may be partly responsible for the antiinflammatory function.

Inhibition of inducible nitric oxide synthase by Acanthopanax senticosus extract in RAW264.7 macrophages.

AIM OF THE STUDY: The herb Acanthopanax senticosus (Siberian ginseng) has long been used as a traditional medicine. However, little is known about anti-inflammatory effects and its mechanisms of action. Excess production of nitric oxide (NO) is one of the characteristics of inflammation. In this study we examined the effects of A. senticosus extract (ASE) on NO production and inducible nitric oxide synthase (iNOS) gene expression in lipopolysaccharide (LPS) plus interferon-gamma (IFN-gamma)-stimulated RAW264.7 macrophages and investigated its mechanisms of anti-inflammatory activity. MATERIALS AND METHODS: RAW264.7 macrophages were treated with 10 microg/ml LPS plus 20U/ml IFN-gamma in the presence or absence of ASE. NO production and iNOS gene expression were investigated. We further evaluated the effect of ASE on oxidative stress-sensitive transcription nuclear factor-kappa B (NF-kappaB) activation. RESULTS: ASE significantly suppressed NO production and iNOS gene expression in a dose-dependent manner. ASE also reduced DNA-binding activity of NF-kappaB in LPS plus IFN-gamma stimulated RAW264.7 macrophages. Further studies indicated that LPS plus IFN-gamma-induced inhibitory factor-kappa B alpha (I-kappaBalpha) degradation and p65 nuclear translocation were inhibited in RAW264.7 macrophages exposed to ASE. Moreover, ASE inhibited the LPS plus IFN-gamma mediated increase in intracellular peroxides production. CONCLUSIONS: These results suggest ASE suppresses iNOS gene expression through the inhibition of intracellular peroxides production, which has been implicated in the activation of NF-kappaB.

Effects of traditional Chinese medicine on immune responses in abalone, Haliotis discus hannai Ino.

A traditional Chinese medicine (TCM) preparation was formulated from orange peel (Pericarpium Citri Reticulatae), hawthorn (Crataegus pinnatifida), astragalus (Astragalus membranaceus (Fisch.) Bunge), pilose asiabell root (Radix codonopsis), indigowoad root (Radix isatidis), taraxacum (Herba taraxaci) and malt (Fructus Hordei Germinatus) at a weight ratio of 1:1:1.5:1.5:1.5:1.5:2. A feeding experiment was conducted to determine the effects of TCM on innate immunity of abalone, Haliotis discus hannai Ino. Artificial diets containing 1%, 3%, 5% TCM preparation, 1% hawthorn or 1% astragalus, respectively, were fed to juvenile abalone (initial weight 10.38+/-2.51 g; initial shell length 44.15+/-4.15 mm) for 80 days. A TCM-free diet was used as a control. Each diet was fed to three replicate groups of abalone using a randomized design. The results indicated that phagocytic activity was significantly higher in abalone fed 3%, 5% TCM preparation, 1% astragalus or 1% hawthorn (P<0.05). Respiratory burst activity was significantly higher in abalone fed 1%, 3%, 5% TCM preparation, 1% astragalus or 1% hawthorn (P<0.05). Agglutination titre was significantly higher in abalone fed 5% TCM preparation (P<0.05). Weight gain ratio (WGR), daily increment in shell length (DISL), total haemocyte count (THC), plasma protein concentration, and the activity of acid phosphatase (ACP) were not significantly affected by the TCM preparation (P>0.05). These results indicate that TCM preparation can modulate the immunity of H. discus hannai, and it is very possible that TCM might be used as immunostimulants in abalone farming.

Acanthopanax senticosus inhibits nitric oxide production in murine macrophages in vitro and in vivo.

Excess nitric oxide (NO) production has been implicated in inflammatory diseases. The present study investigated the inhibitory effect of the stem bark extract of Acanthopanax senticosus (A. senticosus) on NO production in murine macrophages in vitro and in vivo. In vitro exposure of RAW264.7 cells to 1, 10, 50, 100, 250, 500 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by lipopolysaccharide (LPS) and interferon gamma (IFN-gamma) in a dose-dependent manner. In vitro exposure of mouse resident peritoneal macrophages to 1, 10, 100 and 1000 microg/mL of A. senticosus extract significantly suppressed NO production induced by LPS and IFN-gamma in a dose-dependent manner. In vivo administration of A. senticosus extract (50, 100 and 200 mg/kg) to KM mice dose-dependently inhibited LPS and IFN-gamma induced production of NO in isolated mouse peritoneal macrophages ex vivo. Exposure to A. senticosus extract had no effect on cell viability and systemic toxicity. The results demonstrated that the stem bark extract of A. senticosus extract inhibits NO production in murine macrophages in vitro and in vivo.