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.

Application of concentrated deep sea water inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice.

BACKGROUND: Mineral water from deep-sea bedrock, formed over thousands of years, is rich in minerals such as Ca, Mg, Na, K, Fe and others. Our present study was to investigate the preventive effects of natural deep-sea water on developing atopic dermatitis (AD). METHODS: We elicited AD by application of DNCB (2,4-dinitro-chlorobezene) in Nc/Nga mouse dorsal skin. Deep Sea water (DSW) was filtered and concentrated by a nanofiltration process and reverse osmosis. We applied concentrated DSW (CDSW) to lesions five times per week for six weeks, followed by evaluation. 1% pimecrolimus ointment was used as positive control. The severity of skin lesions was assessed macroscopically and histologically. Levels of inflammatory mediators and cytokines in the serum were detected by Enzyme-linked immunosorbent assay (ELISA) and the levels of CD4+ and CD8+ spleen lymphocytes were determined by flow cytometry analysis. RESULTS: DNCB-treated mice showed atopic dermatitis-like skin lesions. Treatment of mice with CDSW reduced the severity of symptoms in the skin lesions, including edema, erythema, dryness, itching, and transepidermal water loss (TEWL). Histological analyses demonstrated that epidermal thickness and infiltration of inflammatory cells were decreased after CDSW treatment. Given these interesting observations, we further evaluated the effect of CDSW on immune responses in this AD model. Treatment AD mice with CDSW inhibited up-regulation of IgE, histamine, and pro-inflammatory cytokines in the serum. Also, the CD4+/CD8+ ratio in spleen lymphocyte was down-regulated after treatment with CDSW. Finally, cytokines, especially IL-4 and IL-10 which are important for Th2 cell development, were reduced. CONCLUSIONS: Our data suggests that topical application of CDSW could be useful in preventing the development of atopic dermatitis.

Anti-Wrinkle Efficacy of Edible Bird's Nest Extract: A Randomized, Double-Blind, Placebo-Controlled, Comparative Study.

This study aimed to evaluate skin health's functional improvement, such as wrinkles, elasticity, moisture, and whitening, and safety following the consumption of "edible bird's nest extract" for 12 weeks by women. This single-center, double-blinded, parallel-group, placebo-controlled study included women aged 40-60 years. Our primary purpose was to assess improvement in skin wrinkles, elasticity, and moisture after 12 weeks using an SV700, cutometer, and corneometer, respectively, compared to baseline measurements. Our secondary purpose was to evaluate skin wrinkle, elasticity, and moisture changes at 4 and 8 weeks from baseline using the aforementioned equipment, and measure transdermal water loss and melanin and erythema indexes using a tewameter and mexameter, respectively. Experts performed the visual evaluation of skin wrinkles at 4, 8, and 12 weeks from baseline. The participants were randomly allocated in a 1:1 ratio into the edible bird's nest extract or the placebo group with 43 participants each, where they consumed 100 mg of the extract or placebo, respectively, daily for 12 weeks. The outcomes were measured at every visit. In this study, upon comparing changes in the skin elasticity value between the two intake groups at 12 weeks of ingestion, skin elasticity in the edible bird's nest extract group decreased significantly compared with that in the placebo group. Adverse reactions were absent in both groups. In the case of laboratory test results, changes before and after the ingestion of the extract were within the normal range, thus indicating no clinically significant difference. The edible bird's nest extract was effective in improving skin wrinkles. Moreover, it is beneficial for skin health and can be used as a skin nutritional supplement. Compared with the placebo, the edible bird's nest extract was identified as safe. Clinical Trial Registration: https://cris.nih.go.kr/cris/search/detailSearch.do?search_lang=E&search_page=M&pageSize=10&page=undefined&seq=21007&status=5&seq_group=20330, identifier KCT0006558.

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.

Component analysis of four-part extracts from Chamaecyparis obtusa Endl. by supercritical fluid extraction and anti-inflammatory effect on RAW 264.7cells.

The main purpose of this study was to suggest a solution for inflammation caused by urban daily life based on supercritical extraction of Chamaecyparis obtusa Endl. (C. obtusa). The leaves, cone, wood, and bark of C. obtusa were separately extracted with supercritical extraction. Alpha-pinene, anti-inflammatory active compound of C. obtusa, was analyzed by electrospray ionization-mass spectrometry, gas chromatography, and gas chromatography mass spectrometry from each part of extracts. As a result of the reduction rate of nitric oxide in RAW 264.7 cells, the wood extract exhibited the highest reduction rate was about 45% among extracts from four part of C. obtusa. The wood extract from C. obtusa can be developed as a good anti-inflammatory natural raw material for natural product industry.

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.

Effect of type II collagen extract on immunosuppression induced by methotrexate in rats.

This study investigated the effect of type II collagen extract on SD rats with deteriorated immunity caused by methotrexate. The test samples were dosed once a day for 28 days by gastric gavage at dosage 250 mg/kg and 500 mg/kg after methotrexate treatment, and the changes on body weight, total blood leukocyte numbers, the percentages of B-cells, CD4+ T-cells and CD8+ T-cells in the blood and spleen were observed. The changes on body weight, the total blood leukocyte numbers, the total lymphocyte numbers in the spleen, the ratio of CD4+ and CD8+ T-cells in the blood and spleen were increased significantly in type II collagen extract groups as compared with the control group. According to the above results, type II collagen extract has an effect of increasing immune responses on rats with deteriorated immunity caused by methotrexate.

Acupuncture decreases ischemia-induced apoptosis and cell proliferation in dentate gyrus of gerbils.

BACKGROUND: Acupuncture has been used for the enhancement of functional recovery from various disorders. In the present study, the effect of acupuncture on the apoptosis and new cell proliferation in the hippocampal dentate gyrus of gerbils (n = 25) following transient global ischemia was investigated. METHODS: To determine the level of apoptosis and cell proliferation, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) were employed respectively. RESULTS: In the dentate gyrus of ischemic gerbils, the number of both TUNEL- and BrdU-positive cells (66.01 +/- 2.45/mm(2) and 514.38 +/- 44.90/mm(2)) was significantly increased compared to that of the sham-operated gerbils (11.25 +/- 1.85/mm(2) and 111.47 +/- 10.95/mm(2)). Among the acupuncture (ST36, LI4 or non-acupoint) treated groups, ST36 acupoint treated group showed the most potent apoptosis (20.52 +/- 2.40/mm(2)) and proliferation (159.38 +/- 24.05/mm(2)) suppressive effects ( approximately 70% decreases in both apoptosis and cell proliferation). CONCLUSION: These results may suggest that acupuncture treatment alleviates ischemia-induced apoptosis and presents possible therapeutic potentials in the recovery from ischemic cerebral injury.

Analgesic effects by electroacupuncture were decreased in inducible nitric oxide synthase knockout mice.

BACKGROUND: This study was designed to investigate the involvement of inducible nitric oxide synthase (iNOS) in electroacupuncture (EA)-induced analgesia. METHODS: Two and 100 Hz EA stimulation were applied at acupoint ST 36 (Zusanli) in iNOS knockout mice (n = 28). Needles were inserted 5 mm in depth in ST36. After insertion, the needles were fixed in situ with adhesive tape. EA was applied after the basal threshold determination. The EA parameters were set as follows: constant square wave current output (pulse width: 0.6 ms at 2 Hz and 0.2 ms at 100 Hz) and 2 mA intensities. EA was performed for 30 minutes and tail-flick latencies (TFLs) were evaluated every 15 minutes for 1 hour. RESULTS: In 2 Hz EA stimulation, the tail-flick response (TFR) of wild-type mice for durations of 0, 15, 30, 45 and 60 minutes were 2.70 +/- 0.26, 4.19 +/- 0.37, 4.17 +/- 0.34, 3.57 +/- 0.27 and 3.39 +/- 0.32 seconds of TFLs. Meanwhile, iNOS -/- mice showed 4.10 +/- 0.33, 4.77 +/- 0.24, 5.26 +/- 0.30, 4.48 +/- 0.33 and 5.00 +/- 0.41 seconds of TFLs. In 100 Hz EA stimulation, the TFR of wild-type mice were 3.01 +/- 0.24, 4.67 +/- 0.31, 4.76 +/- 0.25, 4.04 +/- 0.45 and 4.26 +/- 0.30 seconds of TFLs. The iNOS -/- mice were 4.33 +/- 0.16, 5.29 +/- 0.28, 5.06 +/- 0.35, 4.52 +/- 0.17 and 4.80 +/- 0.28 seconds of TFLs. Wild-type mice exhibited 63.9% increase in TFL compared to the baseline after 2 Hz EA, whereas the iNOS knockout mice exhibited 32.9% increase in TFL. The TFL after 100 Hz EA showed similar trends: 66.5% increase in TFL in wildtype mice and 18.3% increase in the iNOS knockout mice. CONCLUSION: The present findings suggested that iNOS may play a crucial role in both low- and high-frequency EA-induced analgesic effects.

Efficacy of combined treatment by scalp and penetration acupunctures with TKM medication (tang) on stroke patients.

BACKGROUND: Traditional Korean medicine (TKM) therapy of scalp and penetration acupuncture has been used for the treatment of stroke in Korea. This study investigated the efficacy of scalp and penetration acupuncture in combination with TKM medication. METHODS: Twenty-four stroke patients were enrolled in the program. Control group (n = 12) received herbal medicines (Baepungtang, Sopungtang or Sosokmyeongtang) and conventional body acupuncture. Treatment group (n = 12) received scalp and penetration acupuncture in addition to herbal medicines and conventional body acupuncture. Improvements in the motor functions were scored by the modified Barthel index (MBI). RESULTS: After 4 weeks of treatment, statistical analysis showed significant improvement in the MBI scores for both groups. Significant difference in the MBI scores between two groups, however, was not observed. CONCLUSION: The supportive effect of scalp and penetration acupuncture in the treatment of stroke needs further investigation.

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 Platycodon grandiflorum on lipopolysaccharide-stimulated production of prostaglandin E2, nitric oxide, and interleukin-8 in mouse microglial BV2 cells.

The roots of Platycodon grandiflorum, which belongs to the Campanulaceae family, have been used as a food material and as a traditional Oriental medicine. The effect of P. grandiflorum against lipopolysaccharide (LPS)-stimulated inflammation was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, reverse transcription-polymerase chain reaction, prostaglandin E(2 )(PGE(2)) immunoassay, nitric oxide (NO) detection assay, and interleurkin-8 (IL- 8) immunoassay on BV2 microglial cells. The aqueous extract of P. grandiflorum was shown to suppress PGE(2 )synthesis and NO production by inhibiting LPS-stimulated cyclooxygenase (COX)-2 activity and expression of inducible NO synthase (iNOS) mRNAs. In addition, the treatment with P. grandiflorum reduced the LPS-induced IL-8 release. These results suggest that P. grandiflorum inhibits PGE(2) and NO production through its suppression of LPS-induced COX-2 and iNOS expression, and also reduces IL-8 secretion by microglial cells.

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.

Evaluation of a Apo-1/Fas promoter polymorphism in Korean stroke patients.

Apoptosis has been implicated in the pathogenesis of neurodegenerative diseases such as stroke and Alzheimer's disease. Apo-1/Fas gene is one of the mediators of apoptosis in stroke. Mval polymorphism is the first polymorphic marker identified in the Apo-1/Fas gene promoter, which was typed by PCR and followed by Mval digestion and gel electrophoresis. DNA isolated from peripheral blood collected from 91 stroke patients and 103 healthy blood donors was used for genotypes of GG, GA and AA by sequence specific primer PCR. Mval polymorphism was examined based on Fas gene promotor region by restriction fragment length polymorphism (RFLP). The Fas-GG genotype was the least frequent in patients with stroke and healthy controls (P = 0.57). In normal Korean controls the Mval polymorphism GA, AA and GG were 48.6%, 34.9% and 16.5%. In stroke patients were 56.2%, 29.6% and 14.2% respectively. And the allelic frequencies of Mval*2 (G) allele were less frequent than Mval*1 (A) allele in patients with stroke and healthy controls (P = 0.76). In normal Korean controls Mval*1 (A) and Mval*2 (G) alleles were 59.2% and 40.8%. In stroke patients were 57.6% and 42.4%, respectively. Our results, pending confirmation in a larger study, indicate that the Fas genotype may not appear to be a risk factor for stroke in Korean stroke patients.

Alcohol and nicotine reduce cell proliferation and enhance apoptosis in dentate gyrus.

It is generally accepted that alcohol and nicotine affect learning ability and memory functions, especially in adolescents. In the present study, the effects of alcohol and nicotine on cell proliferation and apoptosis in the dentate gyrus of young rats were investigated. The results show that cell proliferation is suppressed by alcohol and nicotine. Furthermore, alcohol and nicotine increase the number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells. Based on the results presented in this study, it can be suggested that alcohol- and nicotine-related impairment in learning and memory functions may be due to alcohol- and nicotine-induced suppression of new cell formation and acceleration of apoptosis, especially during adolescence.

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.

Alcohol induces apoptosis in TM3 mouse Leydig cells via bax-dependent caspase-3 activation.

To investigate whether ethanol induces apoptosis in Leydig cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, terminal deoxynuclotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, DNA fragmentation assay, caspase-3 enzyme assay, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis were performed on TM3 mouse Leydig cells. Through morphological and biochemical analyses, it was demonstrated that TM3 cells treated with ethanol at concentrations of 50 and 100 mM exhibit classical apoptotic features. In addition, it was shown that ethanol induces increases in levels of bax and caspase-3 and a decrease in bcl-2 expression. Based on the results, alcohol appears to activate specific intracellular death-related pathways leading to bax-dependant caspase-3 activation and the induction of apoptosis in Leydig cells.

Acupuncture decreases nitric oxide synthase expression in periaqueductal gray area of rats with streptozotocin-induced diabetes.

Acupuncture has been used as a clinical treatment in Oriental medicine for various diseases including diabetes mellitus, one of the most common metabolic disorders in humans. In the present study, the effect of acupuncture on the expressions of neuronal nitric oxide synthase (nNOS) and nitric oxide synthase (NOS) in the dorsolateral periaqueductal gray (DL-PAG) area of rats with streptozotocin (STZ)-induced diabetes was investigated via nNOS immunohistochemistry and nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry. Enhanced expression of nNOS and NOS was detected in the DL-PAG of rats with STZ-induced diabetes, and acupunctural treatment at Zusanli acupoint suppressed the diabetes-induced enhancement in the expression of nNOS and NOS. The present results demonstrate that acupuncture is effective in the modulation of the expression of nNOS and NOS in the DL-PAG under diabetic conditions.

Protective effect of Hypericum perforatum Linn (St. John's wort) against hydrogen peroxide-induced apoptosis on human neuroblastoma cells.

The medicinal plant Hypericum perforatum Linn, commonly known as St. John's wort, has been used as an antidepressant. To investigate whether St. John's wort possesses a protective effect against hydrogen peroxide (H(2)O(2))-induced cytotoxicity in neuronal cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4,6-diamidino-2-phenylindole staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, flow cytometry analysis, DNA fragmentation assay, and caspase-3 enzyme assay were performed on SK-N-MC human neuroblastoma cells. Cells treated with H(2)O(2) exhibited several apoptotic features, while those pre-treated with St. John's wort prior to H(2)O(2) exposure showed a decreased occurrence of apoptotic features. In addition, pre-treatment with St. John's wort inhibited H(2)O(2)-induced increase in caspase-3 enzyme activity. These results suggest that St. John's wort may exert a protective effect against H(2)O(2)-induced apoptosis in human neuroblastoma cells.