Kaori Munakata

Importance of the interferon-α system in murine large intestine indicated by microarray analysis of commensal bacteria-induced immunological changes

BackgroundAlthough microbiota play a critical role in the normal development and function of host immune systems, the underlying mechanisms, especially those involved in the large intestine (LI), remain unknown. In the present study, we performed transcriptome analysis of the LI of germ-free (GF) and specific pathogen-free (SPF) mice of the IQI strain, an inbred strain established from ICR mice.ResultsGeneChip analysis, quantitative real-time RT-PCR, and reconfirmation using bacteria-inoculated GF mice revealed differences in the expression levels of several immune-related genes, such as cryptdin-related sequences (CRS), certain subsets of type 1 interferon (IFN)-related genes, class Ib MHC molecules, and certain complements. LI expressed no authentic cryptdins but predominantly expressed CRS2, 4, and 7. The mRNA levels of IFN-related genes, including Irf7, Isgf3g, Ifit1 and Stat1, were lower in SPF- and flora-reconstituted mice. When an oral IFN-α inducer tilorone analog, R11567DA, was administered to SPF mice, IFN-α was induced rapidly in the LI at 4 h, whereas no IFN-α protein was detected in the small intestine (SI) or blood. In situ hybridization and immunohistochemistry suggested that the IFN-α production originated from Paneth cells in the SI, and portions of lamina proprial CD11b- or mPDCA1-positive cells in the LI.ConclusionThe present study suggests that microbial colonization, while inducing the expression of anti-microbial peptides, results in the down-regulation of certain genes responsible for immune responses, especially for type I IFN synthesis. This may reflect the adaptation process of the immune system in the LI to prevent excessive inflammation with respect to continuous microbial exposure. Further, the repertoire of anti-microbial peptides and the extraordinary role of interferon producing cells in the LI have been found to be distinct from those in the SI.

Maternal stress affects postnatal growth and the pituitary expression of prolactin in mouse offspring

Maternal stress exerts long‐lasting psychiatric and somatic on offspring, which persist into adulthood. However, the effect of maternal stress on the postnatal growth of pups has not been widely reported. In this study, we found that maternal immobilization stress (IS) during lactation resulted in low body weight of male mouse offspring, which persisted after weaning. Despite free access to chow, IS induced maternal malnutrition and decreased the serum insulin‐like growth factor‐1 (IGF‐1) levels in the mothers and in the pups. mRNA expression analysis of anterior pituitary hormones in the pups revealed that growth hormone (GH) and prolactin (PRL), but no other hormones, were decreased by IS. Expression of the pituitary transcription factor PIT1 and isoforms of PITX2, which are essential for the development and function of GH‐producing somatotropes and PRL‐producing lactotropes, was decreased, whereas that of PROP1, which is critical for the earlier stages of pituitary development, was unchanged. Immunohistochemistry also showed a decrease in pituitary PRL protein expression. These results suggest that stress in a postpartum mother has persistent effects on the body weight of the offspring. Reduced PRL expression in the offsprings pituitary gland may play a role in these effects.

A Kampo (traditional Japanese herbal) medicine, Hochuekkito, pretreatment in mice prevented influenza virus replication accompanied with GM-CSF expression and increase in several defensin mRNA levels.

A Kampo medicine, Hochuekkito (TJ-41), with an influenza virus-preventing effect had life-extending effectiveness, and immunological responses other than interferon (IFN)-α release were examined. TJ-41 (1 g/kg) was given to C57BL/6 male mice orally once a day for 2 weeks. Mice were then intranasally infected with influenza virus. After infection, virus titers and various parameters, mRNA levels and protein expression, for immunoresponses in the bronchoalveolar lavage fluid or removed lung homogenate, were measured by plaque assay, quantitative RT-PCR and ELISA. IFN-α and -β levels of TJ-41-treated mice were higher than those of the control. Toll-like receptor TLR7 and TLR9 mRNAs were elevated after infection, but retinoic acid-inducible gene (RIG-1) family mRNA levels, RIG-1, melanoma differentiation-associated gene 5 and Leishmania G protein 2 showed no response in either TJ-41 or control groups. Interferon regulatory transcription factor (IRF)-3 mRNA levels to stimulate type I (α/β) IFN were increased, but IRF-7 did not change. Only granulocyte-macrophage colony-stimulating factor (GM-CSF) after Hochuekkito treatment was significantly elevated 2 and 3 days after infection. The mRNA levels of 7 defensins after infection increased compared to preinfection values. The key roles of TJ-41 were not only stimulation of type I IFN release but also GM-CSF-derived anti-inflammation activity. Furthermore, defensin (antimicrobial peptide) mRNA levels increased by infection and were further enhanced by TJ-41 treatment. Defensin might prevent influenza virus replication.

First Nationwide Attitude Survey of Japanese Physicians on the Use of Traditional Japanese Medicine (Kampo) in Cancer Treatment

The aim of this nationwide survey was to investigate the use of Kampo medicine by Japanese physicians who worked in the core cancer treatment hospitals which were designated by Ministry of Health, Labour and Welfare. Among the 900 physicians surveyed, 92.4% reported having prescribed Kampo medications, of whom 73.5% reported having prescribed them for cancer patients. Despite this high percentage and the finding that only 9.7% of the physicians reported that they considered Kampo medications to be harmful, only 23.1% of the physicians expressed high expectations of the efficacy of Kampo medicine in tumor suppression and the exertion of immunostimulatory action. In contrast, many cancer patients have expressed the belief that Kampo medications can suppress tumor growth, and several studies have reported that they exert immunostimulatory action. To resolve this discrepancy in patient and physician expectations and to clarify the research findings, further research into the effectiveness and harmfulness of Kampo medicine in cancer treatment is warranted.

Prescription of Kampo Drugs in the Japanese Health Care Insurance Program

Kampo medicine or traditional Japanese medicine has been used under Japans National Health Insurance scheme for 46 years. Recent research has shown that more than 80% of physicians use Kampo in daily practice. However, the use of Kampo from the patient perspective has received scant attention. To assess the current use of Kampo drugs in the National Health Insurance Program, we analysed a total of 67,113,579 health care claim records, which had been collected by Japans Ministry of Health, Labour and Welfare in 2009. We found that Kampo drugs were prescribed for 1.34% of all patients. Among these, 92.2% simultaneously received biomedical drugs. Shakuyakukanzoto was the most frequently prescribed Kampo drug. The usage of frequently prescribed Kampo drugs differed between the youth and the elderly, males and females, and inpatients and outpatients. Kampo medicine has been employed in a wide variety of conditions, but the prescription rate was highest for disorders associated with pregnancy, childbirth, and the puerperium (4.08%). Although the adoption of Kampo medicine by physicians is large in a variety of diseases, the prescription rate of Kampo drugs is very limited.

Clinical Data Mining Related to the Japanese Kampo Concept “Hie” (Oversensitivity to Coldness) in Men and Pre- and Postmenopausal Women

“Hie” is a subjective oversensitivity to cold and a condition experienced in 60% of Japanese citizens. The condition of hie has not been documented in Western medicine. However, in Kampo medicine, hie is an important target of treatment, because it has been considered one of the sources of all kinds of diseases. This study aimed to clarify the symptoms and findings associated with hie and contribute to increased precision in hie diagnosis. During 2005-2006, data from interviews of 1691 patients during their initial visit to the Kampo Clinic of Keio University Hospital were analyzed using a classification and regression tree (CART) analysis, a data mining technique. Symptoms and findings characteristic of each group are follows as, postmenopausal women: fatigability, absence of lower abdominal pain, and absence of hot flashes of feet: women with menstruation: leg swelling, knee pain, and abdominal pain; men: insomnia, leg weakness, and absence of excess saliva. From the perspective of Kampo medicine the result suggested that the feature of hie condition in postmenopausal women, women with menstruation, and men is statistically different.

The Difference between the Two Representative Kampo Formulas for Treating Dysmenorrhea: An Observational Study

In Kampo medicine, two different formulas are effective for treating dysmenorrhea—tokishakuyakusan and keishibukuryogan; however, the criteria by which specialists select the appropriate formula for each patient are not clear. We compared patients treated with tokishakuyakusan and those with keishibukuryogan and proposed a predictive model. The study included 168 primary and secondary dysmenorrhea patients who visited the Kampo Clinic at Keio University Hospital. We collected clinical data from 128 dysmenorrhea patients, compared the two patient groups and selected significantly different factors as potential predictors, and used logistic regression to establish a model. An external validation was performed using 40 dysmenorrhea patients. Lightheadedness, BMI < 18.5, and a weak abdomen were significantly more frequent in the tokishakuyakusan group; tendency to sweat, heat intolerance, leg numbness, a cold sensation in the lower back, a strong abdomen, and paraumbilical tenderness and resistance were more frequent in the keishibukuryogan group. The final model fitted the data well. Internally estimated accuracy was 81.2%, and a leave-one-out cross-validation estimate of accuracy was 80.5%. External validation accuracy was 85.0%. We proposed a model for predicting the use of two Kampo formulas for dysmenorrhea, which should be validated in prospective trials.

Mechanism of Action of the Anti-Influenza Virus Active Kampo (Traditional Japanese Herbal) Medicine, Hochuekkito

When the Kampo medicine, Hochuekkito (Hochu), was administered to normal mice for 2 weeks, influenza virus titer was reduced. The mechanism of action of Hochu was examined using the plaque assay method. It was suggested that Hochu may either obstruct the first stage of the infection process (adsorption and entry) or may directly target viral particles. Using the plaque assay method, these 2 modes of action could not be differentiated. Virus RNA in the infected cell was verified by quantitative real-time polymerase chain reaction. An equal inhibition effect was obtained when Hochu was preprocessed for normal cells and when they were made to act simultaneously with virus adsorption. The viral load at the cell surface following UV irradiation was higher in the Hochu-administered group as compared with that of the control. Moreover, the affinity of Hochu for the influenza virus was hundred times higher than its affinity for the host cell. The effect of entry obstruction by Hochu was observed via image analysis, where the amount of virus nucleocapsid protein (NP) invading the cell was visualized with FITC-labeled NP antibody. Hochu does not seem to have an effect on nucleic acid synthesis, viral release from infected cells, and on the subsequent second round of infection. In conclusion, Hochu binds to viral particles and forms complexes that can obstruct the entry of influenza virus into cells.

Current Status of Kampo Medicine Learning by Japanese Physicians for Cancer Treatment

a Aoyama Pharmacy, 2-10-9 Minami-Aoyama, Minato-ku, Tokyo 107-0062, Japan b Kanagawa Academy of Science and Technology, 3-2-1 Sakado Takatsu-ku, Kawasaki-shi, Kanagawa 213-0012, Japan c Keio Research Institute at SFC, 5322 Endo Hujisawa-shi, Kanagawa 252-8520, Japan d Shiba daimon Imazu Clinic, 1-1-14 Shibadaimon Minato-ku, Tokyo 105-0012, Japan e Center for Kampo Medicine, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan

Current Problems of the Compound Fee for Herbal Medicine

2006年度に改定された漢方生薬の調剤である湯薬調剤料が適正であるか否かを検討した。調剤時間を指標に湯薬調剤と一般調剤を比較した。1～15日処方で一般調剤が平均4．4分／枚のところ湯薬調剤は平均13．4分／枚と約3倍の時間を要し，30日を超過する処方では一般調剤に比べ約7倍の時間を要した。次に調剤業務1分間あたりの調剤報酬を湯薬調剤と一般調剤で比較検討した。1～15日処方では大きな差はなかったが，15日を超える処方日数の湯薬調剤では一般調剤に比べ，調剤報酬が約1／3から1／5ほど低い結果となった。大学病院における処方日数の変移の調査では，30日を超過する処方件数は2003年度では全体の2．7％であったが，2008年度の件数は約14倍に増えて全体の42％であった。長期投薬が増えつつある現状を考慮すると，湯薬調剤料も内服調剤料同様，処方日数に応じた調剤報酬にする必要がある。