Effectiveness and Safety of Progressive Loading-Motion Style Acupuncture Treatment for Acute Low Back Pain after Traffic Accidents: A Randomized Controlled Trial.

BACKGROUND: Traffic injuries include acute low back pain (LBP) needing active treatment to prevent chronicity. This two-armed, parallel, assessor-blinded, randomized controlled trial evaluated the effectiveness and safety of progressive loading-motion style acupuncture treatment (PL-MSAT) for acute LBP following traffic accidents. METHODS: Based on an effect size of 1.03, 104 participants were recruited and divided in a 1:1 ratio into PL-MAST and control groups using block randomization. Both groups underwent integrative Korean medicine treatment (IKMT) daily; only the PL-MSAT group underwent three PL-MSAT sessions. The outcomes were assessed before and after the treatment sessions and at 1 and 3 months post-discharge. The primary outcome was the difference in the numeric rating scale (NRS) for LBP. The secondary outcomes included a visual analog scale for LBP, leg pain status, the Oswestry disability index, lumbar active range of motion (ROM), quality of life, Patient Global Impression of Change, and Post-Traumatic Stress Disorder Checklist adverse events. RESULTS: In the modified intention-to-treat analysis, 50 and 51 participants were included in the PL-MSAT and control groups. On Day 4, the mean LBP NRS score was 3.67 (3.44-3.90) in the PL-MSAT group, indicating a significantly lower NRS 0.77 (0.44-1.11) compared to 4.44 (4.20-4.68) for the control group (p < 0.001). The PL-MSAT group exhibited greater ROM flexion (-5.31; -8.15 to -2.48) and extension (-2.09; -3.39 to -0.80). No significant differences were found for the secondary outcomes and follow-ups. CONCLUSIONS: Compared with IKMT alone, PL-MSAT plus IKMT showed significantly better outcomes for reducing pain and increasing the ROM in acute LBP.

Inhibitory Effects of Wheat Sprouts Extract on RANKL-Induced Osteoclast Differentiation via Suppressing MAPK and NFATc1 Signaling Pathways.

The maintenance of bone is dependent on both osteoclasts, which break down bone, and osteoblasts, which build new bone. Various bone-related disorders, including osteoporosis, can occur as a result of an imbalance between these two cell types. Prolonged use of currently available bone resorption inhibitors may show side effects. Therefore, developing a novel preventive material which effectively inhibits osteoclast differentiation could be beneficial. This study planned to investigate the inhibitory effect of wheat sprout ethanolic extracts (Saegeumgang [SGG] and Arriheuk [ARH]) on the differentiation of osteoclasts induced by RANKL, as well as the mechanisms why fundamental to these effects. The effects of SGG and ARH on bone resorption and osteoclast differentiation were evaluated using RAW 264.7 cells and assessed through TRAP cell count, pit formation, and activity. The expressions of mRNA and protein were accomplished using western blotting, and reverse transcription quantitative polymerase chain reaction analyses were conducted. SGG and ARH were found to suppress osteoclast differentiation in RANKL-stimulated RAW264.7 cells without causing cytotoxic effects. In addition, treatment with SGG and ARH led to a reduction in the number of cells with positive staining for TRAP and TRAP activity. SGG and ARH treatment dose-dependently decreased the pit area in pit formation assays, showing a notable reduction compared to the pit area created by mature osteoclasts. SGG and ARH inhibited osteoclast activity by 84.9% and 95.7% at 200 µg/mL, respectively. In addition, SGG and ARH suppressed the transcriptional activation of various osteoclast-related genes, such as RANK, NFATc1, cathepsin K, c-Fos, TRAP, matrix metallopeptidase-9, dendritic cell-specific transmembrane protein, ATPase H+ transporting v0 subunit d2, and osteoclast-associated receptor in RAW264.7 cells treated with RANKL. SGG and ARH extracts were found to affect the expression of NFATc1 and genes that are specific to osteoclasts during osteoclast differentiation, suggesting their potential use as functional foods or as therapeutic interventions targeting bone health.

Adenosine Deaminase Inhibitory Activity of Medicinal Plants: Boost the Production of Cordycepin in Cordyceps militaris.

Cordycepin, also known as 3'-deoxyadenosine, is a major active ingredient of Cordyceps militaris with diverse pharmacological effects. Due to its limited supply, many attempts have been conducted to enhance the cordycepin content. As part of this study, eight medicinal plants were supplemented with cultivation substrates of Cordyceps to increase the cordycepin content. Cordyceps cultivated on brown rice supplemented with Mori Folium, Curcumae Rhizoma, Saururi Herba, and Angelicae Gigantis Radix exhibited increased cordycepin content compared to a brown rice control. Among them, the addition of 25% Mori Folium increased the cordycepin content up to 4 times. Adenosine deaminase (ADA) modulates the deamination of adenosine and deoxyadenosine, and the inhibitors have therapeutic potential with anti-proliferative and anti-inflammatory properties. As ADA is also known to be involved in converting cordycepin to 3'-deoxyinosine, the inhibitory activity of medicinal plants on ADA was measured by spectrophotometric analysis using cordycepin as a substrate. As expected, Mori Folium, Curcumae Rhizoma, Saururi Herba, and Angelicae Gigas Radix strongly inhibited ADA activity. Molecular docking analysis also showed the correlation between ADA and the major components of these medicinal plants. Conclusively, our research suggests a new strategy of using medicinal plants to enhance cordycepin production in C. militaris.

Noni fruit extract ameliorates alcohol-induced hangover symptoms by reducing the concentrations of alcohol and acetaldehyde in a Sprague Dawley rat model and a human intervention study.

Various studies have reported that Noni shows various health effects. This study aimed to assess the ability of Noni fruit extract to serve as a single active functional ingredient for the alleviation of hangover symptoms in Sprague Dawley rats and healthy subjects in a single-dose, randomized, double-blind, crossover, placebo-controlled study. The rats were orally administered Noni fruit extract at 50 or 100 mg per kg body weight (B.W.) and HOVENIA. The blood ethanol (EtOH) and acetaldehyde concentrations were significantly lower in the 100 mg per kg B.W. group than in the EtOH group. Alcohol dehydrogenase and aldehyde dehydrogenase activity tended to increase in the 100 mg kg-1 B.W. group. In the human study, 30 subjects received either a placebo or Noni fruit extract (1 g). The Noni fruit extract group showed significantly faster time point at which the maximum concentration (Tmax) of alcohol than in the placebo group. In addition, blood acetaldehyde levels and diarrhea at 40 and 720 min after alcohol intake and the area under the curve between 40 and 60 min of acetaldehyde were significantly decreased in the Noni fruit extract group compared to the placebo group. According to the QUalitative INteraction Trees, subjects who were ≤36 years old who consumed more alcohol (>15 drinks per week) and had a higher total hangover score (>27.5 and 33) presented significantly lower blood acetaldehyde levels and less severe hangover symptoms. These results indicate that Noni fruit extract has the potential to improve hangover symptoms by decreasing alcohol and acetaldehyde levels.

Antifatigue and Anti-Inflammatory Effects of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge Complex Extracts in Physically Fatigued Mice.

Fatigue is a common complaint among people under stress, causing an array of negative effects on physical function. In this study, we investigated the antifatigue and anti-inflammatory effects of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge complex extracts (CAA) using a treadmill stress test in animal models. The mice were administered various doses of CAA (50-200 mg/kg bw per day) once daily for 21 days. After exhaustive treadmill exercise, the running time of CAA-treated mice increased 1.5 times; fatigue-related biochemical parameters, including lactate dehydrogenase (∼30%), creatine kinase (∼20%), and proinflammatory cytokines interleukin (IL)-1ß (∼10%), and IL-6 (∼10%) in the serum and muscle tissue were downregulated compared with those in exercised control mice. This study provides strong evidence for the prevention of CAA-induced inflammatory incidences mediated by the blockade of nuclear factor-κB activation. Collectively, our results indicate that CAA can alleviate symptoms of fatigue in mice as an effective anti-inflammatory agent.

Canavalia gladiata Pod Extract Mitigates Ovalbumin-Induced Asthma Onset in Male BALB/c Mice via Suppression of MAPK.

Asthma is one of the most common inflammatory diseases of the lung worldwide. There has been considerable progress in recent studies to treat and prevent allergic asthma, however, various side effects are still observed in clinical practice. Six-week-old male BALB/c mice were orally administered with either sword bean pod extracts (SBP; 100 or 300 mg/kg) or dexamethasone (DEX; 5 mg/kg) once daily over 3 weeks, followed by ovalbumin sensitization (OVA/Alum.; intraperitoneal administration, 50 µg/2 mg/per mouse). Scoring of lung inflammation was performed to observe pathological changes in response to SBP treatment compared to OVA/Alum.-induced lung injury. Additionally, inflammatory cytokines were quantified in serum, bronchoalveolar lavage fluid (BALF), and lung tissue using ELISA and Western blot analyses. SBP treatment significantly reduced the infiltration of inflammatory cells, and release of histamine, immunoglobulin E, and leukotriene in serum and BALF. Moreover, the therapeutic effect of SBP was also assessed to analyze the inflammatory changes in the lung tissues. SBP markedly suppressed the activation of the MAPK signaling pathway and the expression of key inflammatory proteins (e.g., TNF-α) and Th2 type cytokines (IL-5 and IL-13). SBP was effective in ameliorating the allergic inflammation against OVA/Alum.-induced asthma by suppressing pulmonary inflammation.

Sword Bean (Canavalia gladiata) Pod Exerts Anti-Allergic and Anti-Inflammatory Effects through Modulation of Th1/Th2 Cell Differentiation.

Allergy is an immunoglobulin E (IgE)-mediated process, and its incidence and prevalence have increased worldwide in recent years. Therapeutic agents for allergic diseases are continuously being developed, but side effects follow when used for a long-term use. Therefore, treatments based on natural products that are safe for the body are urgently required. Sword bean (Canavalia gladiata) pod (SBP) has been traditionally used to treat inflammatory diseases, but there is still no scientific basis for its anti-allergic effect. Accordingly, this study investigates the anti-allergic effect and its mechanism of SBP in vitro and in vivo. SBP reduced the nitric oxide production and decreased mRNA and protein expression of inflammatory mediates (inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2)), and inhibited the phosphorylation of nuclear factor kappa B (NF-κB), a major signaling molecule in the inflammatory response. Additionally, SBP extract treatment inhibited phosphatidylinositol-3-kinase/mammalian target of rapamycin (PI3K/mTOR) signaling activity to further inhibit degranulation and allergy mediator generation and control the balance of Th1/Th2 cells, which can induce an allergic reaction when disrupted. Furthermore, the SBP extract exhibited anti-allergic effects in anti-dinitrophenyl IgE-induced RBL-2H3 cells and ovalbumin-treated mice. These findings have potential clinical implications for the treatment as well as prevention of allergic diseases.

Naringenin and Phytoestrogen 8-Prenylnaringenin Protect against Islet Dysfunction and Inhibit Apoptotic Signaling in Insulin-Deficient Diabetic Mice.

It has been shown that citrus flavanone naringenin and its prenyl derivative 8-prenylnaringenin (8-PN) possess various pharmacological activities in in vitro and in vivo models. Interestingly, it has been proposed that prenylation can enhance biological potentials, including the estrogen-like activities of flavonoids. The objective of this study was to investigate the anti-diabetic potential and molecular mechanism of 8-PN in streptozotocin (STZ)-induced insulin-deficient diabetic mice in comparison with naringenin reported to exhibit hypoglycemic effects. The oral administration of naringenin and 8-PN ameliorated impaired glucose homeostasis and islet dysfunction induced by STZ treatment. These protective effects were associated with the suppression of pancreatic ß-cell apoptosis and inflammatory responses in mice. Moreover, both naringenin and 8-PN normalized STZ-induced insulin-signaling defects in skeletal muscles and apoptotic protein expression in the liver. Importantly, 8-PN increased the protein expression levels of estrogen receptor-α (ERα) in the pancreas and liver and of fibroblast growth factor 21 in the liver, suggesting that 8-PN could act as an ERα agonist in the regulation of glucose homeostasis. This study provides novel insights into the mechanisms underlying preventive effects of naringenin and 8-PN on the impairment of glucose homeostasis in insulin-deficient diabetic mice.

Ameliorative Effect of Citrus junos Tanaka Waste (By-Product) Water Extract on Particulate Matter 10-Induced Lung Damage.

Citrus junos Tanaka (CJ)-related products are well-accepted by consumers worldwide; thus, they generate huge amounts of waste (peel, pulp, and seed) through CJ processing. Although some CJ by-products (CJBs) are recycled, their use is limited owing to the limited understanding of their nutritional and economic value. The exposure to particulate matter (PM) increases the risk of respiratory diseases. In this study, we investigated the ameliorative effects of CJB extracts (100, 200 mg/kg/day, 7 days) on PM10-induced (10 mg/kg, intranasal, 6 h) lung damage in BALB/c mice. Cell type-specific signaling pathways are examined using the A549 (PM10, 200 µg/mL, 6 h) and RAW264.7 (LPS, 100 ng/mL, 6 h) cell lines. The CJB extracts significantly attenuated PM10-induced pulmonary damage and inflammatory cell infiltration in a mouse model. The essential protein markers in inflammatory signaling pathways, such as AKT, ERK, JNK, and NF-κB for PM10-induced phosphorylation, were dramatically reduced by CJB extract treatment in both the mouse and cell models. Furthermore, the CJB extracts reduced the production of reactive oxygen species and nitric oxide in a dose-dependent manner in the cells. Comprehensively, the CJB extracts were effective in reducing PM10-induced lung injuries by suppressing pulmonary inflammation, potentially due to their anti-inflammatory and antioxidant properties.

Olfactory Stimulation with Volatile Aroma Compounds of Basil (Ocimum basilicum L.) Essential Oil and Linalool Ameliorates White Fat Accumulation and Dyslipidemia in Chronically Stressed Rats.

We explored the physiological effects of inhaling basil essential oil (BEO) and/or linalool and identified odor-active aroma compounds in BEO using gas chromatography/mass spectrometry (GC-MS) and GC-olfactometry (GC-O). Linalool was identified as the major volatile compound in BEO. Three groups of rats were administered BEO and linalool via inhalation, while rats in the control group were not. Inhalation of BEO for 20 min only reduced the total weight gain (190.67 ± 2.52 g) and increased the forced swimming time (47.33 ± 14.84 s) compared with the control group (219.67 ± 2.08 g, 8.33 ± 5.13 s). Inhalation of BEO for 5 min (392 ± 21 beats/min) only reduced the pulse compared with the control group (420 ± 19 beats/min). Inhalation of linalool only reduced the weight of white adipose tissue (5.75 ± 0.61 g). The levels of stress-related hormones were not significantly different among the groups. The total cholesterol and triglyceride levels decreased after inhalation of BEO for 20 min (by more than -10% and -15%, respectively). Low-density lipoprotein cholesterol levels were lowered (by more than -10%) by the inhalation of BEO and linalool, regardless of the inhalation time. In particular, BEO inhalation for 20 min was associated with the lowest level of low-density lipoprotein cholesterol (53.94 ± 2.72 mg/dL). High-density lipoprotein cholesterol levels increased after inhalation of BEO (by more than +15%). The atherogenic index and cardiac risk factors were suppressed by BEO inhalation. Animals exposed to BEO and linalool had no significant differences in hepatotoxicity. These data suggest that the inhalation of BEO and linalool may ameliorate cardiovascular and lipid dysfunctions. These effects should be explored further for clinical applications.

Occurrence and risk characterization of non-nutritive sweeteners in selected food products from Korea.

Non-nutritive sweeteners (NNS) are food additives that impart a sweet taste to food product with lower calories. Acesulfame potassium, aspartame, sodium saccharin, sucralose, steviol glycosides and enzymatically modified stevia are permitted in Korea. The study established the method of each NNS and applied it to each food items consumed in Korea. For risk assessments, the estimated daily intake (EDI) value for each NNS was calculated. EDI values of NNS were compared directly with each ADI (acceptable daily intake). The total estimated daily intake ranges by age compared with the % ADI were 0.12-0.53, 0.93-1.68, 0.05-0.20, 0.06-0.42 and 0.17-0.98% for acesulfame potassium, sodium saccharin, aspartame, sucralose and sum of stevioside and rebaudioside A, which were based on the overall averages. It can be concluded that the daily dietary intake of each of the five NNS is at a safe level when considered as a proportion of the ADI. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-021-01012-9.

Beneficial Effects of a Combination of Curcuma longa L. and Citrus junos Against Beta-Amyloid Peptide-Induced Neurodegeneration in Mice.

Both Curcuma longa (CL) and Citrus junos Tanaka (CJ) have been used to treat various diseases due to their anti-inflammatory and antioxidative stress activities. In this study, we investigated the ameliorative effect of the combination of CL extract and CJ extract (CCC) against beta-amyloid (Aß) peptide-induced neurological damage. CCC prevented neurocytotoxicity in vitro. In addition, it was confirmed that abnormal alternation behavior and memory impairment caused by Aß peptide were reversed by treatment with CCC. Furthermore, CCC treatment led to recovery of the cholinergic system and reactive oxygen species (ROS) oxidative damage defense system. CCC induced expressions of cyclic-adenosine monophosphate response element-binding protein (CREB)-responsive element-binding protein and brain-derived neurotrophic factor were confirmed as was the significantly improved processing in the hippocampus of the mouse Aß peptides. Accordingly, these results suggest that CCC can prevent and/or reverse neurocytotoxicity and cognitive deficits in a mouse model of Alzheimer's disease.

Immature sword bean pods (Canavalia gladiata) inhibit adipogenesis in C3H10T1/2 cells and mice with high-fat diet-induced obesity.

BACKGROUND: Sword bean (SB; Canavalia gladiata) is a perennial vine used as a food and medicinal plant in Asia. SB is rich in nutrients, such as flavonoids and urease, and has various functions, including beneficial effects on dysentery, nausea, and hemorrhoids, as well as anti-inflammatory and antioxidant activity. Various plant parts are used; however, little is known about the physiological effects of SB pods (SBP). In this study, the anti-obesity effects of SBP extract were evaluated. METHODS: To investigate the anti-obesity effects of SBP extract, we confirmed the SBP extract downregulated lipogenesis-related genes and upregulated genes involved in lipolysis and brown adipocyte markers in differentiated C3H10T1/2 adipocytes in vitro. Next, we use a high-fat diet (HFD)-induced obesity mouse model to determine the anti-obesity effects of SBP extract. RESULTS: Treatment with SBP extract significantly reduced adipocytes. The extract decreased the HFD-induced increases in body weight and plasma triglyceride levels in mice after 8 weeks. mRNA and protein levels of the adipogenesis and lipogenesis-related factors CCAAT/enhancer binding protein-ß, CCAAT/enhancer binding protein-α, peroxisome proliferator-activated receptor-γ (PPARγ), and their target genes Ap2, SREBP-1c, FAS, and SCD-1 were reduced by SBP extract. In contrast, AMP-activated protein kinase and sirtuin1, involved in the thermogenic catabolism of fat, were activated by SBP extract in adipocytes and white adipose tissue, increasing the expression of peroxisome proliferator-activated receptor gamma coactivator-1α, peroxisome proliferator-activated receptor-α (PPARα), and uncoupling protein 1 and activating thermogenic activity. CONCLUSION: SBP extract exerts an anti-obesity effect by inhibiting lipogenesis-related factors and activating fat-catabolizing factors; it is, therefore, a promising functional food and natural anti-obesity agent.

Improvement of Fatigue Symptoms and Endurance Capacity by the Combined Administration of Cervus elaphus L., Angelica gigas Nakai, and Astragalus membranaceus Bunge.

Fatigue is a common phenomenon usually observed in healthy, as well as in nonhealthy, individuals that affects their performance and quality of life. Efficient supplementation to relieve fatigue is of significant importance. This study was designed to investigate the efficacy of three prescreened natural resources (Cervus elaphus L. [CEL], Angelica gigas Nakai [AGN], and Astragalus membranaceus Bunge [AMB]) against fatigue symptoms induced by heavy exercise. Effects on muscle fatigue and endurance capacity during exercise were investigated in C2C12 myoblasts and exercised mice. A combination of CEL, AGN, and AMB (CEL:AGN:AMB, 1:2:1) treatment in myoblasts reduced intracellular reactive oxygen species levels induced by hydrogen peroxide by ∼20 times (P < .001). The optimal mixture extract combination was determined as CEL:AGN:AMB, 1:2:1 (CAA), which was recombined by applying the extraction yield of individual substance for in vivo study. Compared to the exercise control (EC) group, the serum lactate dehydrogenase level decreased by ∼40% due to CAA administration. The proliferator-activated receptor gamma coactivator 1-alpha protein expression increased significantly (P < .05) after CAA administration compared to that observed in the normal control group. In parallel, CAA treatment significantly (P < .05) enhanced the maximum running time compared to the EC group. Overall, combinatorial administration exhibited greater efficacy compared to each individual treatment, indicating that CAA could be used as an efficient ergogenic and antifatigue supplement.

Effect of herbal extracts on peripheral nerve regeneration after microsurgery of the sciatic nerve in rats.

BACKGROUND: Recent experimental studies using herbal extracts have shown the possibility of peripheral nerve regeneration. This study aimed to investigate the effects of herbal extracts on peripheral nerve regeneration in a rat sciatic nerve injury model. METHODS: A total of 53 rats were randomly assigned to a control group or one of four experimental groups. In all rats, the sciatic nerve was completely severed and microscopic epineural end-to-end neurorrhaphy was performed. Normal saline (2 mL) was topically applied to the site of nerve repair in the control group, whereas four different herbal extracts - 2 mL each of Astragalus mongholicus Bunge, Coptis japonica (Thunb.) Makino, Aconitum carmichaelii Debeaux, or Paeonia lactiflora Pall. - were topically applied to the site of nerve repair in each experimental group. Nerve conduction studies were performed at an average of 11.9 weeks after the operation, and conduction velocity and proximal and distal amplitudes were measured. Biopsies were performed at an average of 13.2 weeks after the initial neurorrhaphy. The quality of nerve anastomosis and perineural adhesion to the surrounding soft tissues was macroscopically evaluated. The neuroma size at the site of the neurorrhaphy was microscopically measured, whereas the size of the scar tissue was evaluated relative to the diameter of the repaired nerve. RESULTS: The nerve conduction study results showed the highest nerve conduction velocity in the experimental group that used the Coptis japonica (Thunb.) Makino extract and the highest proximal and distal amplitudes in the experimental group that used the Aconitum carmichaelii Debeaux extract. Macroscopic evaluations after the second operation showed that grade 2 perineural adhesion was found in 70.8% of rats. The mean neuroma size in the Coptis japonica (Thunb.) Makino, Aconitum carmichaelii Debeaux, and Paeonia lactiflora Pall. groups showed statistically significant decreases relative to the control group. The mean scar tissue formation index in the Paeonia lactiflora Pall. group showed a statistically significant decrease relative to the control group. CONCLUSIONS: The peripheral nerve regeneration effect of the herbal extracts was confirmed through decreased neuroma and scar tissue formation.

Fermentation of Chestnut (Catanea crenata Sieb) Inner Shell Enhances Anti-Obesity Effects in 3T3-L1 and C3H10T1/2 Adipocytes.

Chestnut inner shell (CIS) is rich in phenols and flavonoids such as gallic acid and ellagic acid, which are known to exhibit effective antioxidant and anti-obesity properties. Fermentation using lactic acid bacteria can enhance the physiological activity by increasing the contents of such functional ingredients. In this study, we evaluated the anti-obesity effects of a CIS extract subjected to a fermentation process (fermented CIS [FCIS]). Treatment with CIS and FCIS extracts (125, 250, and 500 µg/mL) increased cell viability and did not induce apoptosis, indicating no toxicity. The extract suppressed the gene expression of adipogenic factors, peroxisome proliferation-activated receptor gamma, CCAAT/enhancer binding protein (C/EBP) alpha, and C/EBP beta (by 7.75% and 67.59%, 21.41% and 66.27% in 500 µg/mL, respectively), and consequently suppressed the expression of downstream lipogenic factors such as fatty acid synthase, stearoyl CoA desaturase-1, citrate synthase, and ATP citrate lyase. The expression of factors involved in fat catabolism and ß-oxidation increased in a dose-dependent manner, thereby preventing fat accumulation. This observation was consistent with the significant decrease in the staining intensity for lipid droplets, which indicated that lipid accumulation was decreased by 15.46% and 29.44% in 3T3L-1 and 27.01% and 46.68% in C3H10T1/2. Together, these results demonstrate the higher anti-obesity effects of FCIS extract than that of CIS extract, indicating the potential applicability of FCIS as an effective natural raw material to curb obesity.

Development of a Liquid Chromatography/Mass Spectrometry-Based Inhibition Assay for the Screening of Steroid 5-α Reductase in Human and Fish Cell Lines.

Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography-tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC50 values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC50 of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC50 against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.

Mustard Leaf Extract Suppresses Psychological Stress in Chronic Restraint Stress-Subjected Mice by Regulation of Stress Hormone, Neurotransmitters, and Apoptosis.

Mustard leaf (Brassica juncea var. crispifolia L. H. Bailey) has been reported to have psychological properties such as anti-depressant activities. However, studies on chronic stress and depression caused by restraint have not been conducted. Therefore, this study aimed to evaluate the effects of a mustard leaf (ML) extract on chronic restraint stress (CRS) in mice. Male mice were subjected to a CRS protocol for a period of four weeks to induce stress. The results showed that the ML extract (100 and 500 mg/kg/perorally administered for four weeks) significantly decreased corticosterone levels and increased neurotransmitters levels in stressed mice. Apoptosis by CRS exposure was induced by Bcl-2 and Bax expression regulation and was suppressed by reducing caspase-3 and poly (ADP-ribose) polymerase expression after treatment with the ML extract. Our results confirmed that apoptosis was regulated by increased expression of brain-derived neurotrophic factor (BDNF). Additionally, cytokine levels were regulated by the ML extract. In conclusion, our results showed that the ML extract relieved stress effects by regulating hormones and neurotransmitters in CRS mice, BDNF expression, and apoptosis in the brain. Thus, it can be suggested that the studied ML extract is an agonist that can help relieve stress and depression.

Anti-Inflammatory Effect of Immature Sword Bean Pod (Canavalia gladiata) in Lipopolysaccharide-Induced RAW264.7 Cells.

Sword bean has been known as a traditional medicinal plant to treat cancer, sinus infection, and suppurative disease. It also possesses hypertension-relieving, antioxidation, and antibacterial effects. However, studies on the efficacy of sword bean are limited to mature beans. Few studies have focused on immature sword bean pod (ISBP). Therefore, this study aimed to investigate the anti-inflammatory effect of ISBP in RAW264.7 cells stimulated with lipopolysaccharide (LPS). After LPS-induced RAW264.7 cells were treated with ISBP at concentrations (0.5, 1, 2, and 5 mg/mL), levels of nitrite oxide (NO) and prostaglandin E2 (PGE2) production, protein, and mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), inflammatory cytokine secretion level, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activity were determined. Under inflammatory conditions induced by LPS, ISBP reduced levels of inflammatory mediators NO and PGE2 by 60% and 23%, respectively. It also decreased protein and mRNA expression levels of iNOS and COX-2 known to synthesize inflammatory mediators. Inflammatory cytokines, interleukin (IL)-6, and IL-1ß, levels were decreased, while interferon gamma level was increased by ISBP based on enzyme-linked immunosorbent assay (ELISA) and real time-polymerase chain reaction results. Finally, ISBP showed the ability to inhibit NF-κB activity. In conclusion, ISBP can alleviate inflammation by controlling inflammation-related substances, and may have efficacy as a healthful functional food and natural anti-inflammatory drug.

Neuroprotective Activity of Polyphenol-Rich Ribes diacanthum Pall against Oxidative Stress in Glutamate-Stimulated HT-22 Cells and a Scopolamine-Induced Amnesia Animal Model.

Ribes diacanthum Pall, a native Mongolian medicinal plant, has been reported to show antioxidant activities due to its polyphenol and flavonoid content, and is especially rich in the ethyl acetate fraction from an 80% methanol extraction (RDP). We assessed the cytoprotective effect of RDP on glutamate-caused oxidative stress and apoptosis in mouse hippocampal neuronal cells (HT-22 cells). Cell viability was significantly recovered by RDP treatment. Also, RDP effectively decreased the glutamate-induced production of intracellular reactive oxygen species (ROS). In flow cytometric analysis, apoptotic cells and the mitochondrial membrane potential were suppressed by RDP. In the Western blotting analysis, we found that RDP not only decreased the release of apoptotic proteins but also recovered anti-apoptotic protein. Additionally, RDP enhanced the antioxidant defense system by regulating the expression of antioxidant enzymes. Furthermore, treatment with RDP activated the BDNF/TrkB pathway. In accordance with the in vitro results, RDP meliorated memory deficit by defending hippocampal neuronal cells against oxidative damage in scopolamine-injected mice. Taken together, our present study showed that RDP exerted antioxidant and neuroprotective actions against oxidative stress. Therefore, RDP might facilitate the development of candidates for functional health foods for neurodegenerative disorders.