薬理と治療

Glycyrrhizin（GL）is a major active ingredient in licorice root（Glycyrrhiza glabra） with a variety of immunobiological activities. This review describes（ⅰ）the effects of GL on the infections with viruses, bacteria and fungi in various cultures and in immunocompromised animals;（ⅱ）inhibitory effects of GL on the activities of various type 2 cells（PMN-II, type 2 T cells, M2 macrophages）; and（ⅲ）inhibitory mechanisms of action of GL on the M2bMφ polarization in a variety of immunocompromised animals. M2Mφ are known as cells to be responsible on the increased susceptibility of hosts to various opportunistic pathogens. In the latest knowledge, reduced long noncoding RNA Gas5 and increased miR-222 expression inMφ influenced by the γ-irradiation are shown to be associated with M2bMφ polarization. The expression of CCL1, a gene required for M2bMφ survival, is silenced in the γ-irradiated mice due to the Gas5 RNA, which is increased in Mφ after the suppression of miR-222（a Gas5 RNA expression inhibitor）by the GL. Also, the expression of miR︱222 is induced by high-mobility group box 1 （HMGB1）, and GL is known as an inhibitor of HMGB1 binding to DNA in living cells. Based on these facts, the future prospective and usefulness of GL will be discussed.

Background　The parameters used for evaluating the renal function of patients are the measured glomerular filtration rate（GFR）, estimated glomerular filtration rate （eGFR）, and estimated creatinine clearance（CCr）. However, little is known about which of these parameters physicians use to evaluate renal function and how they determine prescription doses in an actual clinical setting. Methods　CCr levels were calculated for 249 patients who had been prescribed levofloxacin at our hospital. Based on these CCr levels, the patients were divided into proper administration group and deviant administration groups. Moreover, eGFR levels and unadjusted eGFR levels were also calculated, and these renal function values were then compared between the two groups to examine whether any differences arose compared with the CCr-based evaluation. Results　In the end, 189 patients （75.9％）were categorized into the proper administration group and 60（24.1％）into the deviant administration group. Meanwhile, the proper administration rates, as evaluated based on the eGFR and unadjusted eGFR levels, were 83.1％ and 77.9％, respectively. Overall, the evaluation results based on the eGFR and unadjusted eGFR levels were inconsistent with the CCr-based evaluation results for 34 （13.6％）and 25（10.0％） patients, respectively. Conclusion　The results of this study indicate that physicians are highly likely to determine doses for prescriptions based on the eGFR levels displayed on electronic medical charts, thereby indicating that the evaluation of patientsʼ renal functions may not be accurate.

Background　We have recently demonstrated the antidepressant effect of liver hydrolysate （LH）. However, the effect of LH on inflammation remains unclear. Therefore, we investigated whether LH has any anti-inflammatory effects on inflammation induced by lipopolysaccharide （LPS）in RAW 264.7 macrophages. Methods　We initially examined the injury caused by various concentrations of LH to the cells. Cell viability was determined by WST-1 assay. To investigate the effect of LH on the mRNA expression of the pro-inflammatory cytokine, we studied the mRNA expression of the pro-inflammatory cytokine, IL-1β by quantitative RT-PCR. To elucidate the mechanism underlying the anti-inflammatory property of LH, we examined whether LH inhibits the NF-κB pathway. The expression of phosphorylated IKKβ and phosphorylated p65 subunit of NF-κB were measured by western blotting. Results　LH treatment distinctly suppressed the expression of IL-1β in comparison to that of the LPS-stimulated cells. The phosphorylation of IKKβ induced by LPS was distinctly reduced by LH treatment. Additionally, the phosphorylation of p65 induced by LPS was distinctly reduced by LH treatment. Conclusion　The results of this study indicated that LH decreased the inflammation induced by LPS in RAW 264.7 cells, at least partially, by suppressing the NF-κB signaling pathway. We propose that the use of LH could be a possible anti-inflammatory strategy.

Objectives　The presented study aimed to investigate whether spermatozoa treated with water-soluble yeast-fermented garlic extract, a novel processed garlic food, affect fertilization and early embryogenesis. Methods　Spermatozoa and oocytes were collected from BALB/cAJcl male mice（12-week-old）and superovulated C57BL/6 J female mice（4-week-old）, respectively. Spermatozoa were pre-treated with water-soluble extraction of yeast fermented garlic, and then used for in vitro fertilization（IVF）. On the following day, the rates of fertilized 2-cell stage embryos, unfertilized oocytes and dead oocytes to total oocytes used for IVF were calculated. After a further 3 days, rates of blastocysts, 2-cell stage embryos and dead embryos to 2-cell stage embryos were calculated. Results　Spermatozoa treated with yeast-fermented garlic extract did not affect fertilization potential such as the number of fertilized 2-cell stage embryos, unfertilized eggs, and dead eggs, but significantly decreased the percentages of 2-cell block during mouse early embryogenesis, leading to significant increase of early embryos that developed up to the blastocyst stage. Conclusions　Yeast-fermented garlic food, a novel processed food, promotes early embryogenesis beyond 2-cell block of mouse embryos, which may be potential seeds for improving IVF in infertility treatment.

Objective　In this study, we investigated the inhibitory effect of Liquid of Fermented Plant Extract SW contained low molecular collagen（hereinafter referred to as LFPE-SW）, a mixture of various fruits and vegetables, on melanogenesis using B16 melanoma cells. Methods　The antioxidant capacity of LFPE-SW was evaluated based on its scavenging ability against superoxide radicals, hydrogen peroxide and free radicals. The inhibition of melanin production was assessed from the amount of melanin produced using B16 melanoma cells. Furthermore, the rate of inhibition of tyrosinase activity and tyrosinase gene expression levels were evaluated. Results　LFPE-SW treatment significantly reduced melanin production in melanocytes. Furthermore, LFPE-SW showed scavenging ability against superoxide radicals, hydrogen peroxide and free radicals. LFPE-SW showed a significant tyrosinase inhibitory effect in vitro, while the expression of tyrosinase mRNA in the B16 melanoma cells did not change after LFPE-SW treatment. Conclusions　These findings suggest that the inhibitory effect of LFPE-SW on melanogenesis is due to the inhibition of tyrosinase activity.

Objectives　The aim of the presented study was to examine the effect of Liquid of Fermented Plant Extract SW（LFPE-SW）on human fibroblasts to cellular oxidative stress and collagen gene expression. Methods　Immortalized human fibroblasts （OUMS-36T-1 cells）were cultured at 37℃ in a 5％ CO2 atmosphere in DMEM containing 10％ FBS for 24 h, and then LFPE-SW was added at the concentration ranges of 0.01-10％ for 24 h. The cell survival rate was measured by Cell Counting Kit-8. The cellular reactive oxygen species（ROS）levels were analyzed with fluorescence microscope using CM-H2DCFDA. The gene expression levels of COL1A1 and COL3A1 were quantified using real-time RT-PCR. Results　LFPE-SW showed no cytotoxicity in OUMS-36T-1 cells at the concentration ranges of 0.01-1％ for 24 h, and it significantly decreased cellular ROS levels under normal culture condition. LFPE-SW also prevented H2O2-induced oxidative stress in OUMS-36T-1 cells. In addition, the expression of COL3A1, but not COL1A1, was significantly increased in OUMS-36 T-1 cells treated with LFPE-SW for 24 h. Conclusions　LFPE-SW has the suppressive effect of ROS levels, which might protect human fibroblast cells from oxidative stress. LFPE-SW also increases COL3A1 expression, suggesting that it may affect collagen biosynthesis.

Objective　To investigate the effect of Bifidobacterium longum BB536-containing synbiotics on the improvement of bowel movements and immune function in the elderly. Methods　A randomized, open-label, parallel-group study was performed on 93 Japanese individuals（aged≥65 years）. Three groups were formed（high dose ［HD］group［taking three sticks of synbiotics/day］, normal dose［ND］group［taking one stick of synbiotics╱day］, and nonintake group）. The intervention period was 4 weeks. Evaluation items comprised Constipation Assessment Scale（CAS-MT）, defecation status, subjective symptoms, and immunity and saliva tests. Results　Per-protocol set analysis included 30, 28, and 29 subjects in the HD, ND, and nonintake groups, respectively. After the four-week intervention, the CAS-MT scores were significantly lower in the ND group than in the nonintake group（P＜0.01）. Meanwhile, by subgroup analysis of subjects with preintake-period scores≥5, the CAS-MT scores of ND and HD groups were significantly lower than the nonintake group（ND group P＜0.01, HD group P＜0.05, respectively）. In the subjective symptom assessment, the question“feeling constipated” was significantly improved in both the ND and HD groups compared with the nonintake group （ND group P＜0.01, HD group P＜0.05, respectively）. In the immunity test, the number of NK cells was significantly increased in the ND group（P＜0.05）and the number of CD8＋T cells was significantly increased in the HD group（P＜0.01）compared with the nonintake group. No adverse events were observed. Conclusions　Continuous consumption of Bifidobacterium longum BB536-containing symbiotics for 4 weeks may contribute to improving bowel movements and immune function in the elderly.

Background/Objectives　Lactoferrin is a multifunctional glycoprotein present in mammalian milk. In the present study, we investigated the effect of 250 mg lactoferrin on abdominal visceral fat in healthy subjects without metabolic syndrome.Methods　In a randomized, double-blind, placebo-controlled, parallel-group trial, healthy Japanese volunteers with a body mass index（BMI）of ≥23 and ＜30 were enrolled and randomly assigned to two groups. For 12 weeks, the subjects in each group took either enteric-coated tablets of lactoferrin coated with alginic acid or placebo. The effects of lactoferrin were evaluated by measuring abdominal visceral fat, subcutaneous fat, total fat, body weight, BMI, waist circumference, and hip circumference as outcomes. This study was registered with UMIN Clinical Trials Registry as UMIN000031545. Results　According to the analysis of subjects who had high visceral fat levels, the amount of changes in abdominal visceral fat, subcutaneous fat, total fat, body weight, and BMI observed during a 12-week intake period was significantly decreased in the lactoferrin group compared with that in the placebo group. No adverse events related to lactoferrin intake were observed in either group. Conclusions　We found that 250 mg lactoferrin decreased abdominal visceral fat in healthy adults, with no adverse effects. Thus, lactoferrin might lead to a reduction in the risk of MetS.

Objectives　The aim of this study was to investigate the effect of oral intake of Amazake using the sake lees and rice koji on intenstinal microbiota on slight constipation patients. Methods　This randomized placebo-controlled crossover-group comparison study included 25 female adult slight constipation patients. Subjects were divided into 13 subjects who drink 125 mL of test beverage（Group Y）and 12subjects who drink 190 g of test beverage（Group A）. In each group, after ingestion of the test beverage or placebo for 30 days, the other beverage was ingested after a 14-days washout period. The feces were collected on the day before and on the last day of ingestion of each beverage, and changes in the intestinal flora due to ingestion were analyzed. Results　In group Y, no significant difference was observed after ingestion of test beverage compared with placebo, but in groupA, significant increase of Bifidobacterium was observed after ingestion of test beverage which include sake lees and rice koji compared with placebo. Conclusions　Ingestion of amazake beverages containing sake lees and rice koji increased the Bifidobacterium in the feces of subjects with slight constipation. Therefore, it is considered that an amazake beverage containing sake lees and rice koji has a function of improving human intestinal bacterial flora. （UMIN-CTR000038323）

Background　Ursolic acid in seaberry exhibits many biological activities. It has been shown to suppress carbachol-induced bladder muscle contraction and TGF-β-induced bladder smooth muscle cell contraction in collagen gel. We previously performed a clinical trial of ursolic acid in Japanese subjects with mild urinary dysfunction. The findings obtained showed that an 8-week treatment with ursolic acid（0.36 mg/day）improved several Kingʼs Health Questionnaire （KHQ）scores, including“Does your bladder problem make you depressed?”. A significant improvement was observed in the overactive bladder syndrome score（OABSS）for the item “How often do you have a sudden desire to urinate, which is difficult to defer?”. However, that study included healthy Japanese subjects with higher OABSS who were suspected as patients. Objective　To adopt the study to the Food with Function Claims system in Japan, we performed a stratified analysis by excluding healthy subjects suspected as patients according to the guidelines of the system. Methods　We excluded subjects who scored 2 points or more to the question“How often do you have a sudden desire to urinate, which is difficult to defer?”for OABSS and 3 points or more for total OABSS. The intervention period was 8 weeks, and urinary symptoms were evaluated by KHQ and OABSS. Results　The results of the stratified analysis showed that the daily intake of ursolic acid （0.36 mg╱day）for 8 weeks improved scores for“Impact of incontinence” and “Emotions”in KHQ. Improvements were also observed in several items related to urinary anxiety by the same dosing protocol. In OABSS, the score for urinary urgency was improved by the 8-week administration of ursolic acid（0.36 mg╱day）. Conclusions　The intake of ursolic acid（0.36 mg╱day for 8 weeks）improved several emotional parameters related to urinary dysfunction. Therefore, ursolic acid in seaberry is useful for reducing moderate urinary symptoms in healthy Japanese subjects. （UMIN000023486）

Objectives　The aim of this study was to investigate the effect of a dietary supplement containing pyroglutamyl tetrapeptide（pEPYP）on knee joint functions in healthy subjects with knee join pain and discomfort. Methods　A total of 42 subjects were randomly divided into either pEPYP（15 mg/day）or placebo group. They orally consumed the respective test substance every day for 12 weeks. The knee joint functions including pain and discomfort were evaluated using JKOM（Japanese Knee Osteoarthritis Measure）and JOA（Japanese Orthopaedic Association）score. This study was registered in UMIN-CTR（ID: UMIN000035610）. Results Compared with the placebo group, pEPYP group showed a significant improvement in the pain score measured by JKOM at week 12（P＝0.048）. Conclusions　These results suggested that the daily supplementation of pEPYP is effective for improving QOL in subjects with knee pain and discomfort.

Objective　This study was conducted to assess the effect of glucose-containing jelly drink intake on cognitive function in healthy adults. Methods　A randomized, double-blind, placebo-controlled, cross-over comparison study was conducted in 16 healthy adult volunteers aged between 20 and 39 in a fasting condition. The subjects performed the Cognitrax Test, which is based on the CNS Vital Signs test battery, 15 min after consuming jelly drink containing 30 g glucose or placebo. Sequential blood glucose was also monitored after oral intake of the jelly drink on a different day. Results　Executive function and motor speed scores were significantly improved after ingestion of the glucose-containing jelly drink compared to the placebo jelly drink. Blood glucose increase due to glucose-containing jelly consumption was observed during the Cognitrax Test. Conclusion　These results suggest that the intake of 30 g glucose-containing jelly drink could improve cognitive function in healthy adults. （UMIN-CTR000039097）

Background　Eyestrain is a serious quality of life problem, especially in the workplace. In particular, work at visual display terminals（VDTs）is a common cause of eyestrain. Monoglucosyl rutin（MGR）is a highly water-soluble α-glucose adduct of rutin. Previous studies have demonstrated that rutin improved dry eye. This randomized, placebo-controlled, double-blind, crossover study was aimed at evaluating the efficacy of oral mono-glucosyl rutin against eyestrain caused by exposure to a visual display terminal（VDT）. Method　This study was designed as a randomized, placebo-controlled, double-blind, crossover study. The participants included thirty healthy Japanese men and women who tended to feel eyestrain. During the first test period, the participants were asked to drink either a test food containing 254 mg of MGR or a placebo food without MGR. After the food intake, the participants did personal computer work at VDT for 30 min and then relaxed for 20 min. We evaluated the participantsʼ eyestrain using an objective parameter （high -frequency component 1, HFC1）and subjective parameters（visual analog scale）. After 7 days, for the second test period, the participants consumed an alternate food following the same protocol as during the first test period. Results　Twenty-seven participants finished the protocol. The HFC1 values for the dominant eye in the monoglucosyl-rutin and placebo groups were 50.25±1.45 and 51.87±1.30, respectively. The subjective parameter of eyestrain was improved in the MGR group in comparison with those in the placebo group. Conclusion　MGR supplementation has the effect to improve eyestrain symptoms due objectively and subjectively to VDT use. （UMIN Clinical Trials Registry ID: UMIN000034224）

Objective　The aim of this study was to investigate the effect of oral intake of food containing piceatannol on fat burning in healthy subjects. Methods　A randomized, double-blind, placebo-controlled crossover comparison study was conducted in 12 healthy subjects. Subjects consumed test food containing 10 mg of piceatannol per day or placebo food for 7 days before and after the washout period, and breath gas was measured. Outcomes of fat burning were evaluated using a respiratory gas analyzer, and respiratory quotient（RQ）was determined during sedentary and low-intensity exercise. Results　Nine out of 12 subjects were included in the final analysis. Compared with that of placebo, oral intake of test food containing piceatannol significantly decreased RQ during sedentary and low-intensity exercise. No change in energy expenditure was observed between the test food and placebo. Conclusions　These results suggested that oral intake of food containing piceatannol increased fat burning during sedentary and low-intensity exercise. （UMIN-CTR ID：UMIN000039427）

Objective　To investigate the effects of citric acid and Moromi vinegar on improving fatigue and relieving psychological stress, we conducted a randomized, double-blind placebo-controlled, parallel-group study in healthy individuals aged≥20 years and ＜65 years who often experienced fatigue from their daily duties. The primary study endpoint was the Visual Analogue Scale（VAS）score for stress, and secondary endpoints included salivary chromogranin A, Profile of Mood States（POMS2）, and Chalder Fatigue Scale score results. Methods　One hundred participants experiencing fatigue were pretested, and 51 healthy individuals without diseases were selected. The 51 healthy individuals were then assigned to either a placebo group, a citric acid（citric acid amount: 810 mg）group, or a Moromi vinegar beverage （citric acid amount: 780 mg）group, and subjected to a 2-week ingestion period, after, stress tolerance test with an Uchida-Kraepelin（U-K）test was administered. Near the time that the stress tolerance test was administered, the VAS score for stress, salivary chromogranin A, and POMS results were evaluated. Additionally, Chalderʼs Fatigue Scale score was evaluated prior to and two weeks after ingestion. The study was registered with the University Hospital Medical Information Network（UMIN）Clinical Trial Registry before implementation （000035299）. Results　Although all participants completed the study, five who ingested＜80％ of the test products were excluded. The VAS stress scores and salivary chromogranin A results in the citric acid and Moromi vinegar beverage groups were significantly different from those in the placebo beverage group 2 weeks after ingestion, and the citric acid and Moromi vinegar-con taining beverages showed a work stress-relieving effect. Moreover, for POMS, there were significant differences in Anger-Hostility between the citric acid and placebo beverage groups 2 weeks after ingestion. There were no significant differences in the Chalder Fatigue Scale scores. Conclusion　Citric acid and Moromi vinegar-containing beverages were shown to reduce stress during mental work in healthy individuals who perceived themselves as susceptible to experiencing daily fatigue.

Objectives　To investigate the sleep-improving effects of oxypinnatanine derived from kwanso （Hemerocallis fulva）, a randomized, double-blind placebo-controlled, parallel-group study was conducted in 32 individuals with sleep disorder who were otherwise healthy. Primary endpoints were the Pittsburgh Sleep Quality Index （PSQI）score, the Oguri-Shirakawa-Azumi （OSA）sleep inventory score, and electroencephalography（EEG）results during sleep. Methods　Among 50 otherwise healthy volunteers, 32 individuals with sleep disorder were selected according to their PSQI scores. During a 2-week pre-observation period, EEG during sleep and OSA sleep questionnaire surveys were conducted. Sixteen participants each were randomized to either ingest food containing oxypinnatanine or placebo over a period of 2 weeks. During week 2 of ingestion, EEG during sleep was measured again. The study was registered with the University Hospital Medical Information Network （UMIN）Clinical Trial Registry before implementation（000035299）. Results All 32 participants completed the study. Analyses of OSA sleep questionnaire responses showed significant differences in sleepiness on rising（factor Ⅰ）, initiation and maintenance of sleep（factor Ⅱ）, and refreshing（factor Ⅲ）for oxypinnatanine versus placebo. Analyses of EEG during sleep also revealed a decrease in the length of sleep interruption and a significant decrease in sleep onset latency for the oxypinnatanine group compared with the placebo group. There was also an increase in deep sleep resulting from a significant increase in the duration of non-rapid eye movement sleep in the oxypinnatanine group compared with the placebo group. Conclusion　Kwanso-derived oxypinnatanine was shown to reduce sleepiness and fatigue on rising and increase deep sleep in healthy individuals with sleep disorder.

Objectives　This study investigated the safety of foods by assessing lipase and α-glucosidase inhibition activities of Terminalia bellirica extract and 12 other functional ingredients. Methods　Lipase and α-glucosidase inhibition activities of T. bellirica extract, oolong tea extract, chitosan, Gymnema sylvestre extract, guava leaves extract, black tea extract, salacia extract, onion skin extract, tea flower extract, du zhong leaves extract, pu︱erh tea extract, green tea extract, and apple extract were determined. Orlistat or acarbose was used as the positive control. Results　Each ingredient displayed lower inhibition activity than orlistat or acarbose. Out of the 13 ingredients, T. bellirica extract displayed the highest lipase inhibition activity in 13 ingredients. Chitosan and salacia extract inhibited α-glucosidase to the same level as T. bellirica extract. Conclusions　It is unlikely that these ingredients cause side effects due to the inhibitory activities. However, the copious use of chitosan and salacia extract with T. bellirica extract may be a concern.