Eight-week supplementation of Aronia berry extract promoted the glutathione defence system against acute aerobic exercise-induced oxidative load immediately and 30 min post-exercise in healthy adults: a double-blind, randomised controlled trial.

BACKGROUND: Food antioxidants have received prompt attention for controlling oxidative stress encountered in daily life. This study aimed to examine the protective effects of Aronia berry extract (ABE) supplementation on acute aerobic exercise (AAE)-induced oxidative stress in healthy subjects. METHODS: We assessed a battery of antioxidant defence and oxidative stress parameters at pre-exercise, immediately post-exercise and 30 min post-exercise in healthy middle-aged adults with habitually low intakes of fruit and vegetables in an 8-week, double-blind, randomised, controlled clinical trial with two arms (n = 70). The AAE challenge model, characterised as a treadmill exercise for 30 min at 60% VO2 maximum, was applied to load oxidative stress at the end of the study. Pearson's correlation analysis assessed the association between the changes in antioxidant defence capacities and oxidative stress levels. RESULTS: The time-course-dependent oxidative stress was well observed in the placebo group regarding the glutathione peroxidase (GPx) activity and the reduced glutathione (GSH) availability for antioxidant defence and erythrocyte malondialdehyde, interleukin-6 and lactate levels for oxidative damage. Meanwhile, the ABE supplementation effectively strengthened the glutathione defence system by increasing GSH availability and GPx activity immediately post-exercise and 30 min post-exercise. In addition, the scatter plot and linear regression analysis revealed strong negative correlations of GSH availability with oxidised low-density lipoprotein and plasma malonaldehyde levels. CONCLUSION: These findings suggest that daily supplementation of 300 mg ABE might help boost GSH levels and an adaptive antioxidant enzyme defence system of erythrocytes in healthy adults with habitually low fruit and vegetable intakes.

KGR-BG1, a Standardized Korean Black Ginseng Extract, Has No Significant Effects on Head or Face Temperature Compared with Korean Red Ginseng Extract and a Placebo.

There is a lack of studies on the effects of Korean ginseng (Panax ginseng C.A. Meyer) on face or body temperature. Therefore, in this study, we evaluated the effects of a black ginseng extract, KGR-BG1, on head and face temperatures and compared them with those of red ginseng extract and a placebo. We assessed their safety and tolerability and examined changes in the serum levels of biomarkers associated with immune responses, as well as with glucose and lipid metabolism. A randomized, double-blind, placebo-controlled study was conducted with 180 participants. The participants were randomly assigned to the KGR-BG1, red ginseng extract, or placebo group. Each group received a 1500 mg oral dose of their respective substances containing 1000 mg of the active component or placebo twice daily for 6 weeks. After treatment, changes in the head, face, and body temperature were measured, and serum biomarkers were evaluated. A total of 172 participants completed the evaluation after 6 weeks of treatment. No significant differences were observed in the head, face, and body temperatures among the treatment groups. After 6 weeks of treatment, the serum levels of biomarkers associated with inflammation, glucose metabolism, and lipid metabolism were similar to the baseline levels in all treatment groups. KGR-BG1 was well-tolerated compared with red ginseng extract and placebo. KGR-BG1 did not significantly alter head, face, or body temperature, or serum biomarker levels, and it was well tolerated in healthy volunteers over 6 weeks of treatment. Study Registration: Registered at Clinical Research Information Service (CRIS; https://cris.nih.go.kr) as KCT0003126.

A novel nutritional mixture, MBN, prevents memory impairment via inhibiting NLRP3 inflammasome formation in 5xFAD transgenic mice.

OBJECTIVES: Amyloid beta (Aß)-induced abnormal neuroinflammation is recognized as a major pathological factor of Alzheimer's disease (AD), which results in memory impairment. Inhibition of excessive neuroinflammation mediated by Aß is considered a promising strategy to ameliorate AD symptoms. To regulate the inflammatory response, nutritional and dietary supplements have been used for centuries. Based on this idea, we investigated whether MBN, a novel nutritional mixture including cassia bark, turmeric root, and ginkgo leaf, can prevent AD progression through neuroinflammatory regulation. METHODS: MBN (10, 30, or 100 µg/ml) and Aß1-42 monomer were incubated together, and the degree of Aß aggregation was measured using Thioflavin T assay. The effects of MBN on Aß pathology in vivo were evaluated by orally administering MBN (40 mg/kg/day for 16 weeks) to five familial AD (5xFAD) mice. RESULTS: We found that treatment with MBN inhibited Aß aggregation in vitro. Next, MBN treatment significantly inhibited the activation of microglia induced by aggregated Aß in 5xFAD mice. Caspase-1 activation, which plays an important role in the maturation of interleukin-1ß, was markedly reduced by MBN. We also found that oral administration of MBN in 5xFAD mice alleviated memory decline. Taken together, our findings demonstrate that MBN suppresses neuroinflammation by downregulating the caspase-1 expression, thereby ameliorating memory impairment in 5xFAD mice. DISCUSSION: Based on these results, we suggest that MBN may be a preventive and therapeutic supplement for AD through the regulation of neuroinflammation.

A Pharmacokinetic Study of Ephedrine and Pseudoephedrine after Oral Administration of Ojeok-San by Validated LC-MS/MS Method in Human Plasma.

A sensitive and reproducible liquid chromatography-tandem mass spectrometry (LC-MS/MS) system was developed and fully validated for the simultaneous determination of ephedrine and pseudoephedrine in human plasma after oral administration of the herbal prescription Ojeok-san (OJS); 2-phenylethylamine was used as the internal standard (IS). Both compounds presented a linear calibration curve (r2 ≥ 0.99) over a concentration range of 0.2-50 ng/mL. The developed method was fully validated in terms of selectivity, lower limit of quantitation, precision, accuracy, recovery, matrix effect, and stability, according to the regulatory guidelines from the U.S. Food and Drug Administration and the Korea Ministry of Food and Drug Safety. This validated method was successfully applied for the pharmacokinetic assessment of ephedrine and pseudoephedrine in 20 healthy Korean volunteers administered OJS.

Pharmacokinetic Analysis and Biomarker-Assisted Safety Assessment of Acetaminophen in Combination With Ojeok-san Compared With Acetaminophen Alone.

Acetaminophen and Ojeok-san are both frequently used analgesics. In this study, we evaluated acetaminophen pharmacokinetics (PK) and changes in microRNA-122 (miR-122) levels after multiple dosing of acetaminophen with or without Ojeok-san. An open-label, 1-sequence, 2-period, 2-treatment crossover study was conducted in 18 subjects. In period 1, 500 mg of acetaminophen was administered 3 times on day 1 and once on day 2. In period 2, after the administration of 14.47 g of Ojeok-san twice on day 2 and 3 times daily on days 3 to 7, Ojeok-san and acetaminophen were coadministered 3 times each on day 8 and once each on day 9. The geometric mean ratios (90% confidence intervals) of acetaminophen with Ojeok-san to acetaminophen alone were 0.98 (0.87 to 1.10) and 1.02 (0.98 to 1.05) for the maximum plasma concentration (Cmax ) and the area under the plasma concentration-time curve during the dosing interval (AUC0-τ ), respectively, of acetaminophen at steady state. The alanine aminotransferase (ALT) levels were within the reference range in all the participants throughout the study period, although the mean fold changes in both serum miR-122 and ALT levels from baseline tended to increase on days 2 to 5. In conclusion, the PK properties of acetaminophen were not significantly affected by Ojeok-san coadministration. For osteoarthritis patients taking acetaminophen with or without Ojeok-san, monitoring potential liver toxicity using miR-122 as a biomarker may be useful.

Study of Pharmacodynamic and Pharmacokinetic Interaction of Bojungikki-Tang with Aspirin in Healthy Subjects and Ischemic Stroke Patients.

BACKGROUND: Bojungikki-tang (BJIKT) is a widely used traditional herbal formula in China, Japan, and Korea. There have been reports that several herbs among BJIKT have interactions with antiplatelet drugs, such as aspirin. This study aimed to assess whether BJIKT interacts with aspirin in terms of pharmacokinetics (PK) and pharmacodynamics (PD) in healthy subjects and ischemic stroke patients. METHODS: The phase I interaction trial was a randomized, open-label, crossover study of 10 healthy male subjects, and the phase III interaction trial was a randomized, placebo-controlled, parallel study of 43 ischemic stroke patients. Each participant randomly received aspirin + BJIKT or aspirin + placebo. For PK analysis, plasma acetyl salicylic acid (ASA) and salicylic acid (SA) were evaluated, and, for PD analysis, platelet aggregation and plasma thromboxane B2 (TxB2) were measured. RESULTS: In the PK parameters, mean area under curve, maximum concertation, and peak concentration time of ASA and SA were not different between two groups in healthy subjects and ischemic stroke patients. In the PD profiles, TxB2 concentrations and platelet aggregation were not affected by coadministration of BJIKT in healthy subjects and ischemic stroke patients. CONCLUSIONS: These results suggest that coadministration of BJIKT with aspirin may not result in herb-drug interaction.

Effects of Ojeok-san on the Pharmacokinetics of Celecoxib at Steady-state in Healthy Volunteers.

Ojeok-san is a frequently used herbal medication for the management of osteoarthritic pain. We evaluated the effect of Ojeok-san on the pharmacokinetics of celecoxib at steady-state in healthy individuals. An open-label, fixed-sequence, two-period, two-treatment cross-over study was conducted. In period I, the individuals received celecoxib capsule 200 mg once daily for 4 days. In period II, only Ojeok-san (14.47 g/pack, three times daily) was administered for 4 days, followed by co-administration with celecoxib for 4 days. On the fourth (final) day of administration, Ojeok-san was administered as a single dose. The blood samples for pharmacokinetic evaluation were collected for up to 48 hr after the administration of celecoxib in each study period. Of the 22 enrolled individuals, 20 individuals completed the study. In the presence of Ojeok-san, the systemic exposure of celecoxib was decreased. The geometric mean ratios ([celecoxib + Ojeok-san]/celecoxib) and the 90% confidence intervals for the maximum plasma concentration (Cmax ) and the area under the plasma concentration-time curve during dosing interval (AUCτ ) of celecoxib at steady-state were 0.725 (0.620-0.848) and 0.885 (0.814-0.962), respectively. The changes in the mean of the Cmax and AUCτ of celecoxib were greater in intermediate metabolizers of cytochrome 2C9 (CYP2C9) than in normal metabolizers. Our results suggested that the Cmax and AUCτ of celecoxib were reduced by Ojeok-san co-administration. This finding may be beneficial to determine the required adjustment of celecoxib dosage when co-administered with Ojeok-san.

Grape Seed Proanthocyanidin Inhibits Mucin Synthesis and Viral Replication by Suppression of AP-1 and NF-κB via p38 MAPKs/JNK Signaling Pathways in Respiratory Syncytial Virus-Infected A549 Cells.

Airway epithelial cells are often infected by respiratory syncytial virus (RSV), one of the most common causes of asthma, bronchiolitis, chronic obstructive pulmonary disease, and pneumonia. During the infection process, excessive mucins instigate airway inflammation. However, the mechanism underlying RSV-induced airway hyper-responsiveness and inflammation is poorly understood. Furthermore, no reliable vaccines or drugs for antiviral therapy are available. In this study, the effect of the natural compound grape seed proanthocyanidin (GSP) on RSV-infected human airway epithelial cells A549 was evaluated. After pretreatment of the cells with or without exposure to RSV with 5-10 µg GSP/mL, the expression of various mucins (MUC1, MUC2, MUC5AC, MUC5B, and MUC8) was evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting, as well as confocal microscopy. We found that GSP significantly decreased RSV-induced mucin synthesis at the mRNA and protein levels. In addition, GSP suppressed the RSV-induced signaling pathways, including extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38, together with nuclear factor kappa B (NF-κB) and activating protein-1 family members (c-Jun and c-Fos). Concomitantly, GSP inhibited the replication of RSV within A549 cells. Taken together, all our results suggest that GSP could be a potent therapeutic agent to suppress excessive mucus production and viral replication in RSV-induced airway inflammatory disorders.

Enhanced Absorption Study of Ginsenoside Compound K (20-O-ß-(D-Glucopyranosyl)-20(S)-protopanaxadiol) after Oral Administration of Fermented Red Ginseng Extract (HYFRG™) in Healthy Korean Volunteers and Rats.

To evaluate the pharmacokinetics of compound K after oral administration of HYFRG and RG in humans, an open-label, randomized, single-dose, fasting, and one-period pharmacokinetic study was conducted. After oral administration of a single 3 g dose of HYFRG and RG to 24 healthy Korean males, the mean (±SD) of AUC0-t and C max of compound K from HYFRG were 1466.83 ± 295.89 ng·h/mL and 254.45 ± 51.20 ng/mL, being 115.2- and 80-fold higher than those for RG (12.73 ± 7.83 ng·h/mL and 3.18 ± 1.70 ng/mL), respectively; in case of Sprague Dawley rats the mean (±SD) of AUC0-t and C max of compound K from HYFRG was 58.03 ± 32.53 ng·h/mL and 15.19 ± 10.69 ng/mL, being 6.3- and 6.0-fold higher than those from RG (9.21 ± 7.52 ng·h/mL and 2.55 ± 0.99 ng/mL), respectively. T max of compound K in humans and rats was 2.54 ± 0.92 and 3.33 ± 0.50 h for HYFRG and 9.11 ± 1.45 and 6.75 ± 3.97 hours for RG, respectively. In conclusion, the administration of HYFRG resulted in a higher and faster absorption of compound K in both humans and rats compared to RG.

Pharmacokinetic comparison of ginsenoside metabolite IH-901 from fermented and non-fermented ginseng in healthy Korean volunteers.

ETHNOPHARMACOLOGICAL RELEVANCE: IH-901 (20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol) is a novel ginseng saponin metabolite formed by human intestinal bacteria and is known to have antitumor and antimetastatic effects. However, there has been no pharmacokinetic study of IH-901 in human beings. AIM OF THE STUDY: The aim of this study was to investigate the pharmacokinetic differences of IH-901 from fermented and non-fermented ginseng. MATERIALS AND METHODS: To investigate whether the pharmacokinetics of IH-901 differ between fermented and non-fermented ginseng, an open label, randomized, single dose, fasting, two-period, cross-over, pharmacokinetic study was conducted. A total of 24 healthy Korean male volunteers participated in this study. All subjects were allocated into two equal groups and administered 3g of fermented or non-fermented Panax ginseng. Serial blood samples for pharmacokinetic analysis were collected in the 24 h after dosing. Plasma IH-901 concentration was measured by a validated high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Pharmacokinetic parameters including AUC(t), C(max), and T(max) were calculated by noncompartmental models in the BA-CALC program (KFDA, 2008, 1.0.0, Korea). RESULTS: After oral administration of fermented ginseng, 5 subjects experienced diarrhea. The means of AUC(t) and C(max) were significantly different between the two groups. In the fermented ginseng group, AUC(t) was 2083.09±91.97 ng h/mL, a 15.5-fold increase over that of IH-901 from the non-fermented group (134.50±63.10 ng h/mL), and the mean C(max) was 325.00±91.97 ng/mL in the fermented ginseng group, a 27-fold higher value than that in the non-fermented group (13.88±7.24 ng/mL). T(max) was 3.29±1.00 and 12.04±4.96 h in the fermented and non-fermented group, respectively. CONCLUSIONS: The results of this study showed that the pharmacokinetic parameters of IH-901 from fermented Panax ginseng are different from those of non-fermented ginseng, from which IH-901 is formed by intestinal fermentation.

Extracts from Citrus unshiu promote immune-mediated inhibition of tumor growth in a murine renal cell carcinoma model.

AIM OF THIS STUDY: Citrus unshiu (Satsuma mandarin, SM) is a citrus fruit the peel of which has been used as a traditional Chinese medicine to treat common cold, relieve exhaustion, and cancer. In this study, we examined how effectively the content and peel extracts of SM can suppress cancer growth. The mechanism underlying cancer-suppressing properties of SM was investigated in tumor-bearing mice with renal carcinoma cell, Renca. MATERIALS AND METHODS: Effectiveness of SM in tumor suppression was evaluated by measuring size of tumor mass in tumor-bearing mice treated with various doses of SM content and peel extracts. Proliferation of tumor cells and splenocytes was determined by MTT assay and [³H]TdR uptake, respectively. Relevant immunological mechanisms were chased by assaying cytokines including TGF-ß, IL-6, IFN-γ, and TNF-α by ELISA. RESULTS: The content and peel extracts of SM inhibited the growth of tumor cells in tumor-bearing mice. Especially, average tumor volume of two groups treated with 3 and 30 mg peel extracts per mouse weight (kg) were significantly decreased to 52.32% (p<0.05) and 68.72% (p<0.01), respectively. To identify tumor regression mechanism, anti-tumor cytokines measured in Con A-activated splenocytes from tumor-bearing mice. IFN-γ was increased in both of the peel extract-treated groups, while TNF-α, which had been decreased by tumor growth, was rescued to the normal level in SM content and peel extracts-treated groups. However, SM content and peel extracts did not inhibit proliferation and tumor-proliferative cytokines including TGF-ß and IL-6 production of tumor cells. CONCLUSION: These results indicate that SM content and peel extracts have anti-tumor properties in the tumor-bearing murine model. The mechanism underlying the anti-tumor effects of SM extracts is strongly suggested to be via boosting cytokines such as IFN-γ and TNF-α, enhancing immune-mediated anti-tumor properties.

Effects of Machilus thunbergii Sieb et Zucc on UV-induced photoaging in hairless mice.

Chronic ultraviolet (UV) exposure induces photoaging and oxidative stress in the skin. We investigated whether Machilus thunbergii Sieb et Zucc (M. thunbergii) could reduce UV-induced photoaging and oxidative stress in hairless mice. The dorsal skin of hairless mice was treated topically with M. thunbergii for 2 h prior to UV irradiation. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were then measured in skin and/or serum samples. Histological changes in the skin were assessed by hematoxylin-eosin (HE) staining. In addition, proteomes from the skin of hairless mice in each group were analyzed. The thickness of the dorsal skin and epidermis was significantly decreased by M. thunbergii treatment. We also found that MDA levels decreased after M. thunbergii treatment and the SOD levels were increased by M. thunbergii compared with those in the UV-only treated group. Proteomic analysis revealed 17 proteins associated with photoaging. These data indicate that M. thunbergii might have antiphotoaging effects.

Bovine colostrum prevents bacterial translocation in an intestinal ischemia/reperfusion-injured rat model.

This study evaluated whether or not bovine colostrum (BC) is able to treat or prevent intestinal barrier damage, bacterial translocation, and the related systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) in an intestinal ischemia/reperfusion (I/R)-injured rat model. Fifty Sprague-Dawley rats were used. The rats' intestinal I/R injuries were induced by clamping the superior mesenteric artery for 30 minutes. After 3 hours of reperfusion and then twice daily reclamping during the experiment, the experimental group was given BC (4 mL/kg/day) perorally, and the other groups received 0.9% saline and low fat milk (LFM) after intestinal I/R injury. Seventy-two hours later we assessed (1) intestinal damage and intestinal permeability, (2) enteric bacterial count and bacterial translocation, (3) serum albumin, protein, and hepatic enzyme levels, (4) pathologic findings of ileum and lung, (5) activity of oxygen-free radical species, and (6) pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta). Intestinal damage, intestinal permeability, and bacterial translocation to other organs were significantly reduced in rats fed with BC after I/R when compared to rats fed LFM/saline after I/R (P < .05). In the evaluation of acute lung injury, neutrophils were found only in the lungs of the saline-fed group after I/R, and the wet/dry ratio of the lung tissue was significantly reduced in the BC-fed group after I/R compared to other I/R groups. A marked difference was found between LFM/saline-fed groups and BC-fed groups regarding malondialdehyde (P < .05) and pro-inflammatory cytokines (P < .01). In conclusion, BC may have beneficial effects in treating and preventing intestinal barrier damage, bacterial translocation and the related SIRS and MODS in the intestinal I/R-injured rat model.

Epigallocatechin gallate stimulates glucose uptake through the phosphatidylinositol 3-kinase-mediated pathway in L6 rat skeletal muscle cells.

The effect of epigallocatechin gallate (EGCG) on glucose uptake was studied in L6 rat skeletal muscle cells. Glucose uptake assay revealed that EGCG increased glucose uptake >70% compared to control. EGCG-stimulated glucose uptake was blocked by LY294002, an inhibitor of phosphatidylinositol (PI) 3-kinase, which is a major regulatory molecule in glucose uptake pathways. However, AMP-activated protein kinase (AMPK), which is another crucial mediator in independent glucose uptake pathways, did not inhibit EGCG-stimulated glucose uptake by SB203585, a specific inhibitor of the AMPK downstream mediator, p38 mitogen-activated protein kinase (MAPK). We also found that EGCG increased the phosphorylation level of protein kinase B and PI 3-kinase activity, when assessed by PI 3-kinase assay, whereas no increase in the phosphorylation level of AMPK and p38 MAPK was observed. Taken together, these results suggest that EGCG might stimulate glucose uptake, not AMPK-mediated but PI 3-kinase-mediated, in skeletal muscle cells, thereby contributing to glucose homeostasis.

Genetic association between 5'-upstream single-nucleotide polymorphisms of PDGFRB and schizophrenia in a Korean population.

PDGFRB is located on chromosome 5q31-q32, a chromosomal region identified by linkage analyses to contain a susceptibility gene for schizophrenia (SCZ). Recent research has focused on the role of the N-methyl-d-aspartate (NMDA) receptor in the pathogenesis of SCZ. D4 dopamine receptor-mediated transactivation of the gene encoding platelet-derived growth factor receptor beta (PDGFRB) has immediate effects on synaptic neurotransmission via calcium-dependent inactivation of NMDA receptors. In this study, we investigate the association between the PDGFRB gene and SCZ in a Korean population. We screened 6 single-nucleotide polymorphisms (SNPs) in the 5'-upstream region of PDGFRB and conducted a case-control study of 381 SCZ patients and 752 controls. The genotype and haplotype frequencies of 3 of the 6 SNPs [SNP1 (g.-1924T>C, rs3756314), SNP3 (g.-1772A>G, rs3756312) and SNP4 (rs3756311, g.-1658G>A)] were significantly associated with SCZ [SNP1, corrected p=0.012 (co-dominant model), 0.002 (Dominant model), and 0.506 (Recessive model); SNP3 and 4, corrected p=0.003, 0.009, and 0.049]. Haplotype analysis also revealed that ht1 (CGG) and ht2 (TAA) were significantly associated with SCZ (ht1, corrected p=0.018, 0.340, and 0.010; ht2, corrected p=0.002, 0.009, and 0.016). Transient transfection in neuronal cells revealed that ht1 had higher luciferase activity than the vector alone. Furthermore, Pdgfrb expression was increased in the frontal cortex and hippocampus in a mouse model of SCZ induced by MK801. We conclude that SNPs of the 5'-upstream region of PDGFRB are associated with SCZ in a Korean population. These are weak positives that require future studies to confirm these results.

Suppressive effects of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression by Citrus reticulata extract in RAW 264.7 macrophage cells.

Immature peels of Citrus reticulata extract (CR) are widely used as traditional herbal medicine in Korea. We studied its effects on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in RAW 264.7 macrophage cells. NO production was assessed by nitrite assay and iNOS expression was identified by reverse transcription-polymerase chain reaction (RT-PCR), Real-time PCR and Western blot. The promoter activity of iNOS gene was also determined by luciferase reporter gene assay using 5'-flanking region of murine iNOS gene. Activation of nuclear factor kappa B (NF-kappaB) was determined by electrophoretic mobility shift assay (EMSA). CR (20, 50, and 100 microg/ml) significantly inhibited the lipopolysaccharide (LPS)-induced NO production (P<0.01; 9.2+/-1.5, 4.8+/-0.6, and 3.3+/-0.4 microM), iNOS protein (38.1+/-3.8, 32.3+/-5.8, and 36.8+/-4.5%) and mRNA expression (34.2+/-4.1, 13.1+/-5.8, and 20.8+/-1.2%) in RAW 264.7 macrophage cells. CR (20, 50, and 100 microg/ml) also reduced the iNOS promoter activity (68.7+/-3.9, 50.6+/-5.6, and 45.9+/-3.9%) in piNOS-LUC-transfected cells. In addition, CR (20, 50, and 100 microg/ml) significantly inhibited the activity of NF-kappaB DNA binding activity in LPS-induced macrophage cells (P<0.05; 51.8+/-4.1, 32.7+/-5.5, and 35.7+/-2.9%). These results suggest that CR may suppress LPS-stimulated NO production by inhibiting of NF-kappaB.

Peroxisome proliferator-activated receptor-gamma Pro12Ala polymorphism is associated with the susceptibility to ischemic stroke in Taeeumin classified by Sasang medicine.

BACKGROUND: Sasang constitutional medicine classifies mankind into four constitutional types according to individual psychologic and physical traits. We hypothesized that differences among constitutional types might be explained by genetic variations. METHODS: To evaluate the hypothesis, we determined the possible association in ischemic stroke patients (n = 134) of peroxisome proliferator-activated receptor (PPAR)-gamma with four constitutional types of Sasang medicine. The constitutional type of each patient and control subject (n = 129) was classified and genotyped for PPAR-gamma polymorphism Pro12Ala by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) methods. RESULTS: The distribution of the Pro/Ala genotypes in the ischemic stroke patients was not significantly different from that of healthy controls [odds ratio (OR)= 0.46; p = 0.1214]. However, very interestingly, we observed that all six Pro/Ala genotypes in ischemic patients were Taeeumin, one of four constitutional types of Sasang medicine. Statistical analysis revealed that Pro/Ala genotype in Taeeumin increases almost 15-fold the susceptibility to ischemic stroke compared to other constitutional types, Taeyangin, Soyangin or Soeumin (OR= 14.72; p = 0.0110). CONCLUSION: From the results in this study, we might suggest that Pro/Ala genotype in Taeeumin is associated with the susceptibility to ischemic stroke. To the author's best knowledge, this is the first report to study on genetic level the potential relationship between ischemic stroke and Sasang constitutional medicine, one of traditional Korean medicines (TKM). Authors hope that this study could provide a new approach for the study of ischemic stroke and merit further research.

Suppressive effect by Hizikia fusiforme on the production of tumor necrosis factor in BV2 murine microglial cells.

BACKGROUND: Hizikia fusiforme has been commonly used as food in Korea. Antioxidant effect of Hizikia fusiforme, however, was recently reported. Thus, herein, we investigated the effect of Hizikia fusiforme on the production and expression of tumor necrosis factor (TNF), a major proinflammatory mediator, in lipopolysaccharide (LPS)-activated BV2 microglial cells. METHODS: Cells were pre-treated with 5 or 50 mug/ml Hizikia fusiforme and treated with 1 mug/ml LPS. The production of TNF was measured by enzyme-linked immunosorbent assay (ELISA). The effect of Hizikia fusiforme on the expression of TNF was also performed by immunoblot analysis and reverse transcription-polymerase chain reaction (RT-PCR). Activation of nuclear factor kappab (NFkappab) was determined by electrophoretic mobility shift assay (EMSA). RESULTS: We observed that Hizikia fusiforme decreased the production of TNF. The inhibitory effect of the Hizikia fusiforme on the expression of TNF was confirmed by immunoblot and RT-PCR analyses. In addition, EMSA experiment revealed that Hizikia fusiforme blocked the LPS-induced activation of NFkappab. CONCLUSION: The present study suggests that Hizikia fusiforme may suppress LPS-stimulated TNF production via inhibition of NFkappab in murine microglial cells.

Ganoderma lucidum extract stimulates glucose uptake in L6 rat skeletal muscle cells.

The effect of Ganoderma lucidum extract on glucose uptake was studied in L6 rat skeletal muscle cells. G. lucidum extract increased glucose uptake about 2-fold compared to control. The extract stimulated the activity of phosphatidylinositol (PI) 3-kinase which is a major regulatory molecule in the glucose uptake pathway. About 7-fold increased activity of a PI 3-kinase was observed after treatment with G. lucidum extract, whereas PI 3-kinase inhibitor, LY294002, blocked the G. lucidum extract-stimulated PI 3-kinase activity in L6 skeletal muscle cells. Protein kinase B, a downstream mediator of PI 3-kinase, was also activated by G. lucidum extract. We then assessed the activity of AMP-activated protein kinase (AMPK), another regulatory molecule in the glucose uptake pathway. G. lucidum extract increased the phosphorylation level of both AMPK alpha1 and alpha2. Activity of p38 MAPK, a downstream mediator of AMPK, was also increased by G. lucidum extract. Taken together, these results suggest that G. lucidum extract may stimulate glucose uptake, through both PI 3-kinase and AMPK in L6 skeletal muscle cells thereby contributing to glucose homeostasis.

Suppressive effect of Punica granatum on the production of tumor necrosis factor (Tnf) in BV2 microglial cells.

While the anti-oxidant properties of Punica granatum methanol extract (PGMF) are well documented, little is known concerning the anti-inflammatory effect of Punica granatum. PGMF was pretreated in BV2 microglial cells and cells were stimulated to induce inflammation by lipopolysaccharide (LPS). The effect of PGME on the production and expression of tumor necrosis factor alpha (Tnf, previously known as Tnf alpha) was determined by enzyme-linked immunosorbent assay (ELISA), western blotting, and reverse transcription-polymerase chain reaction (RT-PCR). In addition, the expression of nuclear factor kappa b (Nfkappab) was measured using an electrophoretic mobility shift assay (EMSA). By ELISA, PGME at the concentrations of 1, 5, 10, and 50 microg/ml inhibited Tnf production in LPS-stimulated cells by 30.2, 42.3, 57.6, and 88.4%, respectively, compared to LPS-stimulated cells. The LPS-stimulated Tnf production was reduced with a dose-dependent manner. Immuno blot and RT-PCR analyses revealed that PGME of 5 and 50 microg/ml inhibited the expression of both protein and mRNA levels of Tnf compared to LPS-stimulated cells. EMSA revealed that PGME of 5 and 50 microg/ml blocked the LPS-stimulated activation of Nfkappab. These data suggest that PGME may suppress LPS-stimulated Tnf production through inhibition of Nfkappab in BV2 microglia cells.