Suramin prevents the development of diabetic kidney disease by inhibiting NLRP3 inflammasome activation in KK-Ay mice

Aims/Introduction Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasomes produce IL-18 upon being activated by various stimuli via the P2 receptors. Previously, we showed that serum and urine IL-18 levels are positively associated with albuminuria in patients with type 2 diabetes, indicating the involvement of inflammasome activation in the pathogenesis of diabetic kidney disease (DKD). In the present study, we investigated whether the administration of suramin, a nonselective antagonist of the P2 receptors, protects diabetic KK.Cg-A(y)/TaJcl (KK-Ay) mice against DKD progression. Materials and Methods Suramin or saline was administered i.p. to KK-Ay and C57BL/6J mice once every 2 weeks for a period of 8 weeks. Mouse mesangial cells (MMCs) were stimulated with ATP in the presence or absence of suramin. Results Suramin treatment significantly suppressed the increase in the urinary albumin-to-creatinine ratio, glomerular hypertrophy, mesangial matrix expansion, and glomerular fibrosis in KK-Ay mice. Suramin also suppressed the upregulation of NLRP3 inflammasome-related genes and proteins in the renal cortex of KK-Ay mice. P2X4 and P2X7 receptors were significantly upregulated in the isolated glomeruli of KK-Ay mice and mainly distributed in the glomerular mesangial cells of KK-Ay mice. Although neither ATP nor suramin affected NLRP3 expression in MMCs, suramin inhibited ATP-induced NLRP3 complex formation and the downstream expression of caspase-1 and IL-18 in MMCs. Conclusions These results suggest that the NLRP3 inflammasome is activated in a diabetic kidney and that inhibition of the NLRP3 inflammasome with suramin protects against the progression of early stage DKD

PDEIV阻害剤を骨粗鬆症治療薬として開発するための基礎的研究

金沢大学大学院自然科学研究科急速に高齢化社会が進行している今日、骨粗髭症の効果的な予防法および治療法を早急に確立する必要がある。我々は、キサンチン誘導体からcyclic AMP-specific phosphodiesterase(PDEIV)に対する選択的阻害剤を誘導し、PDEIV阻害剤の新しい薬理作用として、in vitroにおいてもin vivoにおいても骨芽細胞を活性化して化骨形成を促進することを見いだした。この成果をさらに発展させて以下のような成果を得た。1) キサンチン構造を電子論的に解析し、化学構造をデザインし、合成、試験した結果、既存のPDEIV阻害剤より強く、かつ選択的なPDEIV阻害作用を示す3,4-dipropyl-4,5,7,8-tetrahydro-3H-imidazo[1,2-i]purin 5-One(XT-611)を得た。さらにXT-611は100mg/kgを経口投与しても催吐作用を初めとする副作用を示さないことも確認した。2) PDEIV阻害剤は、in vitroにおいてもin vivoでも、明らかな骨代謝改善作用を有するため新規な骨粗鬆症治療薬として有望であることを示した。3) 骨組織には特有なペプチドのアミノ酸配列を詳細に検討した結果、これらにはいずれも酸性アミノ酸が集中している配列の存在することに着目し、種々検討を重ねた結果、アスパラギン酸(Asp)が6個連続した小ペプチドがハイドロキシアパタイトとの結合親和性が高く、骨組織集積性の高く、骨組織での滞留時間も極めて長いため、骨粗鬆症治療薬の骨組織ターゲッティングのためのキャリアーとして有望であることを示した。4)ラット乳癌Walker256/Sは、自身がLH-RHを過剰に産生・分泌するため、宿主ラットは閉経状態となり、骨粗鬆症様骨変化を起こすことを明らかにした。従って、Walker256/S担癌ラットは、閉経後骨粗髭症のモデルとなること提示した。We previously developed new xanthine derivatives having selective PDE IV inhibitory activity, and suggested that they exhibited osteoblastogenic and a1ti- osteoclastogenic actions. We attemped to further develop new anti-osteoporosis drugs arid osteoporosis animal model in the research project and obtained the following results ;1)A new heterocycle-condensed purine, 3 , 4-dipropyl-4, 5,7, 8-tetrahydro-3H-imidazo [l, 2-i]purin 5-one (XT-611), which shows selective and potent PDE lV inhibitory activity without emetic action, was developed according to the analysis of the structural and electronic properties of alkylxanthine derivatives.2)Our developed compounds and other known PDE IV inhibitors showed significant anabolic actions in the in vitro and in viva experimental systems.3)We newly developed an acidic peptide [(Asp)6], which was a good carrier of drug for bone targeting.4)We showed that Walker256/S mammary carcinoma caused osteoporisis-like changes in rats and ectopically secreted LH-RH, resulting inhibition of sex hormone secretion and stopping the sex cycle. Then, this tumor may be a useful animal model for postmenopausal osteoprosis.研究課題/領域番号:08838025, 研究期間(年度):1996 – 1998出典：研究課題「PDEIV阻害剤を骨粗鬆症治療薬として開発するための基礎的研究」課題番号08838025（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-08838025/088380251998kenkyu_seika_hokoku_gaiyo/）を加工して作

LH-RH遺伝子導入癌細胞による閉経後骨粗鬆症モデル動物の作成

金沢大学医学部・附属病院平成11年度1)ラット乳癌細胞Walker256/S細胞は、PTHrPをはじめとする骨吸収性のホルモンを産生分泌することなしに、LH-RHを過剰分泌しており、このため、脳下垂体からのエストロゲン分泌調節機構が破壊された結果、低エストロゲン血症となる。さらに、腫瘍移植に反応してTNF-αなどの骨吸収性のサイトカインの血中濃度が上昇した。このことから、LH-RH分泌能を有する腫瘍細胞を動物に移植することで骨粗鬆症モデルを作成する本研究目的の妥当性が明らかとなった。2)Walker256/S細胞DNAからLH-RH遺伝子をクローニングし、pBK-CMV発現ベクターに組み込み、まず、ラット腹水肝癌細胞AH66にトランスフェクトしたが、LH-RH蛋白分泌能を有するトランスフェクタントは得られなかった。また、AH66細胞はPTHrPを分泌していることも明らかとなった。平成12年度3)PTHrP非産生細胞を探索したが可移植性動物癌細胞のほとんどがPTHrPを産生分泌していることが明らかとなった。そこで、PTHrP産生能のないことが確認されたマウス培養細胞NIH3T3へのLH-RH関連遺伝子フラグメントの導入を試みた。しかし、遺伝子導入が成功した細胞にあっても当該mRNA発現は認められるものの、LH-RH蛋白を分泌する細胞を得ることはできなかった。4)この原因を追求したところ、NIH3T3細胞はLH-RH切り出し酵素を持たないためであることが示唆された。結果として、本研究の計画年度内に所期の目的を達成することはかなわなかった。しかし、現在は、Walker256が特殊な細胞であるのか、あるいは他の内分泌系の癌細胞の中にLH-RH切り出し酵素を保有しているものがあるかを検討中である。研究課題/領域番号:11877383, 研究期間(年度):1999 – 2000出典：研究課題「LH-RH遺伝子導入癌細胞による閉経後骨粗鬆症モデル動物の作成」課題番号11877383（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11877383/）を加工して作

Phosphodiesterase 4 inhibitor rolipram potentiates the inhibitory effect of calcitonin on osteoclastogenesis

金沢大学医学部附属病院薬剤部To assess the combination effect of calcitonin and the phosphodiesterase 4 inhibitor rolipram on osteoclastogenesis, adherent cell-depleted bone marrow cells from mouse tibia and femur (ACD-BMCs), which were cultured in the presence of 25 ng/ml colony-stimulating factor 1 (CSF-1) and 100 ng/ml soluble receptor activator of NF-κB ligand (sRANKL), were utilized. Calcitonin inhibited formation of tartrate-resistant acid phosphatase-positive multinucleated cells, as mature osteoclasts, by 70% even at 20 pM, whereas rolipram (10 μM) scarcely affected osteoclast formation; in contrast, the combination of both agents led to significant inhibition of multinucleation and pit formation ability of osteoclasts. The combined administration of calcitonin and rolipram attenuated calcitonin receptor mRNA expression in comparison to treatment with either agent alone, whereas expression of RANK and CSF-1 receptor mRNAs was unchanged. Alone, these agents scarcely elevated intracellular cyclic AMP (cAMP) concentration; however, combination treatment with both agents significantly increased cAMP concentration in osteoclast progenitors and osteoclasts. The combination effect was abolished by H-89, an inhibitor of protein kinase A. It appears that rolipram inhibited hydrolysis of cAMP formed by calcitonin in cells and potentiated the inhibitory effect of calcitonin on osteoclastogenesis. The escape phenomenon following calcitonin treatment may also be prevented by concomitant treatment with the phosphodiesterase 4 inhibitor. © Springer-Verlag Tokyo 2006

Issues and Prospect on the Theory of Regional Economics

廣田司朗教授古稀記念特

Affinity selection of DNA-binding protein complexes using mRNA display

Comprehensive analysis of DNA–protein interactions is important for mapping transcriptional regulatory networks on a genome-wide level. Here we present a new application of mRNA display for in vitro selection of DNA-binding protein heterodimeric complexes. Under improved selection conditions using a TPA-responsive element (TRE) as a bait DNA, known interactors c-fos and c-jun were simultaneously enriched about 100-fold from a model library (a 1:1:20 000 mixture of c-fos, c-jun and gst genes) after one round of selection. Furthermore, almost all kinds of the AP-1 family genes including c-jun, c-fos, junD, junB, atf2 and b-atf were successfully selected from an mRNA display library constructed from a mouse brain poly A(+) RNA after six rounds of selection. These results indicate that the mRNA display selection system can identify a variety of DNA-binding protein complexes in a single experiment. Since almost all transcription factors form heterooligomeric complexes to bind with their target DNA, this method should be most useful to search for DNA-binding transcription factor complexes

Investigation of actual use of laxatives and application to the active drug information offer in drug control guidance

金沢大学医学部附属病院薬剤部On the therapy of constipation, there are few reports that surveyed the actual circumstance of patients taking laxatives and the effectiveness of the drug. In this study, we investigated the contents in prescription, the actual conditions of administration of laxatives, and the subjective symptoms of inpatients in all wards of Kanazawa University hospital (2000. 12.1-2000. 12. 10). As a result of the investigation, the percentage of patients prescribed some laxatives was found to be about 31.5% (274) of all hospitalized patients (871), and among them about 16% (43) of the patients had some problems in defecation control. One of the causes leading to the problems for some patients (11) among them seemed to be due to the lesser amount of magnesium oxide taken per day. Then, in the ophthalmology ward we further investigated the proper use of laxatives and the defecation condition of the patient using a questionnaire paper and a record paper (20 cases). Patients became constipated by hospitalization, and although being administered some laxatives, they were often insufficient to control the defecation. Some patients (7) with changeable defecation control could be improved by change or addition of prescription. Pharmacist concerned 5 cases of patients with change or addition of prescription. This study indicates that pharmacists should actively offer information and rational usage of laxatives to doctors and patients

Obesity-induced increase of CYP2E1 activity and its effect on disposition kinetics of chlorzoxazone in Zucker rats

金沢大学医学部附属病院薬剤部This study was designed to investigate the induction of CYP2E1 in obese Zucker rats and its effect on the disposition kinetics of chlorzoxazone (CZX). CZX 20 mg/kg was administered to three groups of rats: normal Zucker rats fed a normal diet (ND), normal Zucker rats fed a high-fat diet (HF), and genetically obese Zucker rats fed a normal diet (OB). The values of the area under the plasma concentration–time curve from 0 to 1 (AUC1) of CZX were in the order of ND > HF > OB rats. The AUC1 values of total 6-hydroxychlorzoxazone (6OHCZX-T), which is considered to be a CYP2E1 metabolic marker, were in the opposite order. The values of the AUC1 ratio (6OHCZX–T/CZX) in ND, HF and OB rats were approximately 0.2, 0.3 and 0.4, respectively. The CZX concentration in fat was much higher than the concentrations in plasma, liver and kidney in all groups. Induction of CYP2E1 protein was greater in both liver and fat of OB rats than in those of HF rats. Microsomal activity of CYP2E1 in liver and fat was also in the order of OB > HF > NMrats. These results suggest that CYP2E1 may be induced in liver and fat of obese patients, thereby potentially altering the disposition kinetics of not only CZX, but also other lipophilic drugs metabolized by CYP2E1.©2006 Published by Elsevier Inc

シュウガクテキ チリョウ ガ ソウコウ シタ シンコウ チョウカンマク アクセイ リンパシュ ノ 1レイ

Mesenteric malignant lymphoma is comparative rare and has a poor prognosis in anadvanced case.Here, we report a patient with intestinal obstruction due to mesenteric malignantlymphoma treated successfully by gastrointestinal bypass operation, chemo-and radiotherapy.A 67-year-old man was admitted to our hospital because of abdominal pain andnausea. An abdominal CT scan revealed a huge tumorous lesion with a soft tissue density,which involved descending colon. Since the symptoms was persisted after conservativetreatment, gastrointestinal bypass procedures were surgically formed. During the laparotomy,an open incisional biopsy was performed. Histological examination showed non-Hodgkinlymphoma with diffuse, large-sized and B-cell type. This case was classified into high-risk groupacccording to International Prognostic Index (IPI). After the operation, 5 courses of combinedchemotherapy (THP-COP) and 30 Gy radiation were performed. These multidisciplinarytreatment is considered to be very effective. Since then complete response and goodpatient’s QOL has been attained for these 3.5 years

6U CubeSat for Ultraviolet Time-Domain Astronomy

A wide-field ultraviolet observatory for time-domain astronomy utilizing 6U CubeSat is presented. Ultraviolet waveband is one of the unexplored fields in astronomy. Potential targets are short duration transient sources in UV-band: early-phase emission from gravitational wave sources, supernovae shock-breakouts, tidal disruption events around super massive blackholes, etc. The telescope was designed for covering the large error circle of GW detectors, FoV~100 deg2. Thanks to the high quantum efficiency of “delta-doping” detector, the detection limit achieves 20 mag (AB) for 1800 s exposure in NUV band, which is sufficient to detect UV emission from a binary neutron star merger within 200 Mpc from the earth. The satellite has a high-performance on-board computer for on-orbit analysis to detect transient sources and measure the magnitude and the accurate position of the target. The obtained information is required to be transferred to the ground within 30 min from the detection to start multi-messenger follow-up observations utilizing ground-based observatories and astronomical satellites. In this presentation we show the mission overview and conceptual design of the satellite system