Antioxidative and anti-inflammatory effect of Phellinus igniarius on RAW 264.7 macrophage cells.

This study investigated the antioxidative and anti-inflammatory effect of Phellinus igniarius (PI) on RAW264.7 mouse macrophages. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Measurement of nitric oxide (NO) synthesis was performed using the NO detection. To identify mRNA expressions of cyclooxygenase-2 (COX-2), inducible NO synthase (iNOS), interleukin (IL)-1α, IL-1ß, IL-6, and tumor necrosis factor (TNF)-α, real time polymerase chain reaction (PCR) was performed. Assessment of prostaglandin E2 (PGE2) synthesis was performed using the PGE2 immunoassay. Measurement of free radical scavenging activity was performed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The MTT assay revealed that PI exerted no significant cytotoxicity in the RAW 264.7 macrophage cells. From the PGE2 immunoassay and NO detection, PGE2 and NO synthesis were significantly suppressed in the PI treated groups compared to the lipopolysaccharide (LPS) treated groups. Real-time PCR analysis revealed that the mRNA expression of COX-2, iNOS, IL-1α, IL-1ß, IL-5, and TNF-α were significantly decreased in the PI treated groups compared to the LPS treated groups. And, PI showed dose-dependent increase in the DPPH radical scavenging activity. In conclusion, PI maybe offer a valuable mode of therapy for the treatment of inflammatory diseases.

Trapa japonica Pericarp Extract Reduces LPS-Induced Inflammation in Macrophages and Acute Lung Injury in Mice.

In this study, we found that chloroform fraction (CF) from TJP ethanolic extract inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and intracellular ROS in RAW264.7 cells. In addition, expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) genes was reduced, as evidenced by western blot. Our results indicate that CF exerts anti-inflammatory effects by down-regulating expression of iNOS and COX-2 genes through inhibition of MAPK (ERK, JNK and p38) and NF-κB signaling. Similarly we also evaluated the effects of CF on LPS-induced acute lung injury. Male Balb/c mice were pretreated with dexamethasone or CF 1 h before intranasal instillation of LPS. Eight hours after LPS administration, the inflammatory cells in the bronchoalveolar lavage fluid (BALF) were determined. The results indicated that CF inhibited LPS-induced TNF-α and IL-6 production in a dose dependent manner. It was also observed that CF attenuated LPS-induced lung histopathologic changes. In conclusion, these data demonstrate that the protective effect of CF on LPS-induced acute lung injury (ALI) in mice might relate to the suppression of excessive inflammatory responses in lung tissue. Thus, it can be suggested that CF might be a potential therapeutic agent for ALI.

Thymol from Thymus quinquecostatus Celak. protects against tert-butyl hydroperoxide-induced oxidative stress in Chang cells.

The present work describes the protective effects of thymol isolated from Thymus quinquecostatus Celak. against tert-butyl hydroperoxide (t-BHP)-induced oxidative damage through various experiments with Chang liver cells. Thymol significantly protected hepatocytes against t-BHP-induced cell cytotoxicity as demonstrated by increased viability. Furthermore, observation of Hoechst staining, annexin V/PI staining, and expression of Bcl-2 and Bax indicated that thymol inhibited t-BHP-induced Chang cell damage. Further, thymol inhibited the loss of mitochondrial membrane potential in t-BHP-treated Chang cells and prevented oxidative stress-triggered reactive oxygen species (ROS) and lipid peroxidation (malondialdehyde, MDA). Thymol restored the antioxidant capability of hepatocytes including glutathione (GSH) levels which were reduced by t-BHP. These results indicated that thymol prevents oxidative stress-induced damage to liver cells through suppression of ROS and MDA levels and increase of GSH level.

Antioxidant activity and protective effects of Trapa japonica pericarp extracts against tert-butylhydroperoxide-induced oxidative damage in Chang cells.

In this study, the antioxidant properties of Trapa japonica pericarp extracts were evaluated through several biochemical assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH), alkyl radical scavenging activity, hydroxyl radical scavenging, ferric reducing antioxidant power (FRAP) assay, ABTS radical scavenging activity and oxygen radical absorbance capacity (ORAC). The antioxidant activities were compared with other natural and synthetic antioxidants. The results showed that higher radical scavenging activity and antioxidant capacity in FRAP than those of vitamin C as a positive control. T. japonica pericarp extracts have antioxidant properties through its ability to prevent tert-butylhydroperoxide (t-BHP)-induced toxicity which enhance the cell viability, reduce reactive oxygen species (ROS) production, inhibits of oxidative damage and mitochondria dysfunction in Chang liver cells. Therefore, based on these finding, it seems reasonable to suggest that T. japonica pericarp extracts has the potential to protect liver against t-BHP-induced cell damage and should be considered as a potential functional food.

In vitro protective effects of Thymus quinquecostatus Celak extracts on t-BHP-induced cell damage through antioxidant activity.

The purpose of this study was to evaluate the antioxidative activities of water and 70% ethanolic extracts from the Thymus quinquecostatus Celak (TQC) for natural antioxidant source. The antioxidant activities were compared with other natural and synthetic antioxidants. The levels of total polyphenols and flavonoids were also determined. The extracts were found to have different levels of antioxidant properties in a few kind of assay. The results showed that higher radical scavenging activity, reducing power and antioxidant capacity in FRAP than those of BHT as a positive control. In addition, the extracts from the TQC leaf and stem showed stronger antioxidant activity than that of vitamin C, α-tocopherol in ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. Cytoprotective and anti-apoptotic effect of water extracts from TQC was also prevented t-BHP-induced toxicity in Chang liver cells. Therefore, these results indicate that TQC extracts have antioxidant properties through its ability to enhance the cell viability, reduction of production of ROS, inhibition of oxidative damage, mitochondria dysfunction and ultimately inhibition of cell apoptosis. Based on the results described above, it is suggested that TQC has the potential to protect liver on t-BHP-induced cell damage and should be considered as a prospective functional food.

Chrysanthemum morifolium Ramat (CM) extract protects human neuroblastoma SH-SY5Y cells against MPP+-induced cytotoxicity.

ETHNOPHARMACOLOGICAL RELEVANCE: Chrysanthemum morifolium Ramat (Asteraceae) has (CM) long been used in Korean and Chinese traditional herbal medicines with numerous therapeutic applications. AIM OF THE STUDY: To evaluate the neuroprotective activities of Chrysanthemum morifolium (CM) extract against 1-methyl-4-phenylpridinium ions (MPP(+)), Parkinsonian toxin through oxidative stress and impaired energy metabolism, in human SH-SY5Y neuroblastoma cells and the underlying mechanisms. MATERIALS AND METHODS: The effects of CM against MPP(+)-induced cytotoxicity and neuronal cell viability, oxidative damage, the expression of Bcl-2 and Bax, caspase-3 and poly(ADP-ribose) polymerase (PARP) proteolysis were evaluated by using SH-SY5Y neuroblastoma cells. RESULTS: CM effectively inhibited the cytotoxicity and improved cell viability. CM also attenuated the elevation of reactive oxygen species (ROS) level, increase in Bax/Bcl-2 ratio, cleavage of caspase-3 and PARP proteolysis. CONCLUSION: These results demonstrate that CM possesses potent neuroprotective activity and therefore, might be a potential candidate in neurodegenerative diseases such as Parkinson's disease.

Inhibitory effects of Angelicae Gigantis Radix on osteoclast formation.

Angelicae Gigantis Radix (AGR) is one of the most widely used herbal medications. AGR is the dried root of Angelica gigas Nakai (Umbelliferae), which is known as Korean angelica. This study investigated the effects of AGR on osteoclast formation using primary bone marrow cells. TNF-alpha treatment increased tartrate-resistant acid phosphatase (Trap) positive cells and Trap activity in bone marrow cells. However, AGR significantly decreased both TNF-alpha-induced Trap positive cells and Trap activity. RT-PCR analyses revealed that AGR decreased mRNA levels of Trap and matrix metalloproteinase-9 in TNF-alpha-treated bone marrow cells. In addition, AGR decreased TNF-alpha-induced activation of NF-kappaB. These results suggest that AGR has an inhibitory effect on the formation of osteoclasts and its effect is partially related to the NF-kappaB pathway.

Protective effects of a novel herbal decoction on focal cerebral ischemia in a rodent model.

BACKGROUND: Herbs have been used to treat stroke and coma patient in traditional Korean medicine (TKM). The novel decoction, Guhpoongchungsimhwan (GCH), was developed on the basis of clinical data and TKM theory. METHODS: We examined the neuroprotective effect of GCH on cerebral ischemia. The middle cerebral artery occlusion (MCAO) model was used to produce cerebral ischemia in Sprague-Dawley rats. Subjects were treated with GCH (50 or 200 mg/kg) or vehicle alone (controls) 0 and 2 hours after MCAO. The functional status was tested 24 hours after MCAO by neurological examination (clinical score) and by series of motor function tasks (foot placement and parallel bar crossing). RESULTS: The infarct volume was determined by 2,3,5-triphenyltetrazolium chloride staining 24 hours after surgery, and the expression of cyclooxygenase-2 was determined by immunohistochemistry. The clinical score of the GCH-treated group (200 mg/kg) was significantly lower than that of the control group (p<0.05), indicating fewer neurological deficits. The impairment of motor functions induced by MCAO was significantly reduced by the administration of GCH (p<0.05). The infarct volume was significantly smaller in the GCH-treated group (203.1 +/- 40.2 mm(3), p<0.05), as compared to the control group (377.8 +/- 32.6 mm(3)). The level of motor function in the GCH-treated group was associated with reduced infarct volume. In the analysis of immunohistochemistry, GCH treatment markedly inhibited the ischemia-induced expression of PTGS2 (prostaglandin-endoperoxidase synthase 2) or cyclooxygenase 2 (COX2), which plays an important role in ischemic neuronal cell death. CONCLUSION: The results showed that GCH reduced the infarct size and the functional deficits in MCAO rats.

Effects of ephedrae herba on melanogenesis and gene expression profiles using cDNA microarray in B16 melanocytes.

Ephedrae Herba (EH) is obtained by drying the stems of Ephedra sinica Stapf (family Ephedraceae). EH has been used clinically to treat colds and to reduce edema in skin. The effects of EH on melanogenesis were studied in B16 murine melanocytes. The tyrosinase activity and melanin content were measured after incubation with EH. Both reverse transcription-polymerase chain reaction and cDNA microarray analysis were used to study the mechanism of EH action. EH decreased tyrosinase activity and melanin content in a dose-dependent manner. EH decreased microphthalmia-associated transcription factor (MITF) and tyrosinase gene expression. These effects were controlled by MITF-mediated regulation of tyrosinase gene expression. EH also altered the expression of about 100 other genes. These results suggest that EH may be used clinically to treat freckles and liver spots.

Melatonin attenuates amyloid beta25-35-induced apoptosis in mouse microglial BV2 cells.

Melatonin has been reported to possess strong antioxidant actions, and is able to directly scavenge a variety of reactive oxygen species (ROS). The present study investigated whether melatonin possesses protective effects against Abeta-induced cytotoxicity in microglial cells. Cells treated with Abeta exhibited several characteristic features of apoptosis, while cells pre-treated with melatonin prior to exposure to Abeta showed a decrease in the occurrence of such apoptotic features. Several previous studies have demonstrated the involvement of ROS in Abeta-induced neurotoxicity, and ROS generated by Abeta have been reported to lead to the activation of nuclear factor-kappa B (NF-kappaB), a transcription factor; pre-treatment with melatonin in the present study reduced the level of Abeta-induced intracellular ROS generation, inhibited NF-kappaB activation, and suppressed the Abeta-induced increase in caspase-3 enzyme activity. In addition, it was found that pre-treatment with melatonin inhibits Abeta-induced increase in the levels of bax mRNA and that it enhances the level of bcl-2 expression. Based on these findings, the authors speculate that melatonin may provide an effective means of treatment for Alzheimer's disease through attenuation of Abeta-induced apoptosis.

Influence of maternal alcohol administration on c-Fos expression in the hippocampus of infant rats.

Maternal alcohol consumption during pregnancy is known to have a detrimental effect on the development of the fetus and its central nervous system (CNS) in particular. In the present study, the dose-dependence of the effect of maternal alcohol on hippocampal c-Fos expression, which is a marker of hippocampal neuronal activity and which is induced by a variety of stimuli, was examined in infant rats. In the present study, it was shown that expression of c-Fos in the hippocampus is decreased following treatment with alcohol in a dose-dependent fashion. Based on the results of the present study and the findings of other studies, it can be suggested that suppression of c-Fos expression in the hippocampus of infant rats with maternal alcohol administration mediates the associated developmental retardation and/or anomalies.

Acupuncture decreases neuropeptide Y expression in the hypothalamus of rats with Streptozotocin-induced diabetes.

It has been known that acupuncture has various effects such as analgesia, promotion of homeostasis, improvements in brain circulation, and rectification of internal disorders. Neuropeptide Y (NPY), a 36-amino-acid peptide, is known to increase appetite. In the present study, the effect of acupuncture stimulation at Zusanli (St.36) on NPY expression in the Streptozotocin (STZ)-induced diabetic rats was investigated via immunohistochemistry. Increased NPY expression was detected in both the Arcuate nucleus (ARN) and the Paraventricular nucleus (PVN) of the Hypothalamus in rats with in STZ-induced diabetes. Needling on Zusanli resulted in decreased NPY levels in both the ARN and PVN of diabetic rats. The present study shows that acupuncture suppressed NPY expression in the ARN and PVN of the Hypothalamus in STZ-induced diabetic rats, suggesting the possibility that acupuncture treatment is effective in curbing the hyperphagia of diabetes.

Atractylodes japonica suppresses lipopolysaccharide-stimulated expressions of inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 macrophages.

Atractylodes japonica has traditionally been used for the treatment of pain and arthritis. The effect of Atractylodes japonica against lipopolysaccharide-induced inflammation was investigated using reverse transcription-polymerase chain reaction (RT-PCR), nitric oxide detection, and prostaglandin E2 (PGE2) immunoassay in mouse RAW 264.7 macrophages. The aqueous extract of Atractylodes japonica suppressed nitric oxide production and PGE2 synthesis by inhibition of the lipopolysaccharide-stimulated enhancement of inducible nitric oxide synthase and cyclooxygenase-2 mRNAs expressions in RAW 264.7 macrophages. These results suggest that Atractylodes japonica exerts anti-inflammatory and analgesic effects probably by suppression of the inducible nitric oxide synthase and cyclooxygenase-2 expressions.

Protective effect of aqueous extract of Ginseng radix against 1-methyl-4-phenylpyridinium-induced apoptosis in PC12 cells.

Ginseng radix, the root of Panax ginseng C. A. MEYER (Araliaceae), is one of the best-known Oriental medicinal herbs with numerous therapeutic applications. To investigate whether Ginseng radix possesses a protective effect against 1-methyl-4-phenylpyridine (MPP(+))-induced cytotoxicity in neuronal cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and caspase-3 enzyme assay were performed on PC12 neuronal cells. Cells treated with MPP(+) exhibited various apoptotic features, while cell pretreated with Ginseng radix prior to MPP(+) exposure showed a decrease in the occurrence of apoptotic features. These results suggest that Ginseng radix may exert a protective effect against MPP(+)-induced apoptosis in PC12 cells.

Acupuncture modulates expressions of nitric oxide synthase and c-Fos in hippocampus after transient global ischemia in gerbils.

The effects of acupuncture on the expressions of nitric oxide synthase (NOS) and c-Fos in the hippocampus of gerbils after transient ischemia were investigated via nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and Fos immunohistochemistry. In animals of the ischemia-induction groups, both common carotid arteries were occluded for 5 minutes. Animals of the acupunctued groups were given acupunctural treatment at Zusanli twice daily for 9 consecutive days. Acupuncture was shown to decrease NADPH-d and c-Fos levels in both the sham-operation group and the ischemia-induction group. These results suggest that acupuncture modulates the expressions of NOS and c-Fos in the hippocampus.

Protective effect of Acanthopanax senticosus against ethanol-induced apoptosis of human neuroblastoma cell line SK-N-MC.

The protective effect of Acanthopanax senticosus (AS) against ethanol (EtOH)-induced apoptosis of the human neuroblastoma cell line SK-N-MC was investigated via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometric analysis, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), and caspase-3 assay. It was shown that cells treated with EtOH exhibit classical apoptotic features, while cells pre-treated with Acanthopanax senticosus prior to EtOH exposure showed decreased occurrence of apoptotic features. In addition, Acanthopanax senticosus pre-treatment was shown to inhibit EtOH-induced increase in caspase-3 mRNA expression and activity. These results suggest that Acanthopanax senticosus may exert a protective effect against EtOH-induced apoptosis of human neuroblastoma cells.

Acupuncture suppresses ischemia-induced increase in c-Fos expression and apoptosis in the hippocampal CA1 region in gerbils.

Acupuncture has been used for the enhancement of functional recovery from various disorders including stroke. In the present study, the effects of acupuncture on the c-Fos expression and apoptosis in the hippocampal CA1 region of gerbils following transient global ischemia were investigated via immunohistochemistry for c-Fos and caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Enhanced Fos, TUNEL, and caspase-3 positivities were detected in the hippocampal CA1 region in the ischemic gerbils. Acupunctural treatment suppressed the ischemia-induced increment in the number of Fos-, TUNEL-, and caspase-3-positive cells: the most potent suppressive effect was observed at the Zusanli acupoint. These results suggest that acupunctural treatment alleviates ischemia-induced apoptosis and may aid in the recovery following ischemic cerebral injury.

Bee venom induces apoptosis and inhibits expression of cyclooxygenase-2 mRNA in human lung cancer cell line NCI-H1299.

To investigate whether bee venom (BV) induces apoptosis, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, terminal deoxynucleotidyl transferase- mediated dUTP nick end-labeling assay, 4,6-diamidino-2-phenylindole staining, flow cytometric analysis, and DNA fragmentation assay were performed on NCI-H1299 lung cancer cells treated with BV. Through morphological and biochemical analyses, it was demonstrated that NCI-H1299 cells treated with BV exhibit several features of apoptosis. In addition, reverse transcription-polymerase chain reaction and prostaglandin E(2) (PGE(2)) immunoassay were performed to verify whether BV possesses an inhibitory effect on the expression of cyclooxygenase (COX) and PGE(2 )synthesis. Expression of COX-2 mRNA and synthesis of PGE(2) were inhibited by BV. These results suggest the possibility that BV may exert an anti-tumor effect on human lung cancer.

Effect of ginseng radix on c-Fos expression in the hippocampus of streptozotocin-induced diabetic rats.

Aqueous extracts of Ginseng radix have traditionally been used in the treatment of diabetes mellitus. In the present study, the effect of Ginseng radix on c-Fos expression in the hippocampus of streptozotocin (STZ)-induced diabetic rats was investigated via immunohistochemistry. Decreased c-Fos expression in the CA regions of the hippocampus was observed in STZ-induced diabetes, and administration of Ginseng radix enhanced the STZ-induced inhibition of c-Fos expression both dose- and duration-dependently. These results suggest that hyperglycemia-induced suppression of Fos expression may trigger the diabetes-induced disruption of hippocampal information processing and that Ginseng radix may alleviate this diabetes-induced disturbance in hippocampal functions.

Nicotine administration decreases neuropeptide Y expression and increases leptin receptor expression in the hypothalamus of food-deprived rats.

The effects of nicotine on the expressions of neuropeptide Y (NPY) and leptin receptor in the rat hypothalamus were investigated via immunohistochemistry. The results show that NPY expression is not affected in the arcuate nucleus (ARN) and is increased only slightly in the paraventricular nucleus (PVN) by nicotine administration under normal (i.e. fed) conditions and that leptin receptor expression is decreased slightly in the ARN and not affected in the PVN following nicotine treatment under the same conditions. Food deprivation enhanced NPY and suppressed leptin receptor expression in the ARN and PVN of the hypothalamus. Nicotine administration resulted in decreased NPY and increased leptin receptor levels.