遺伝性高低比重リポ蛋白コレステロール血症の分子遺伝学的研究

取得学位 : 博士（医学）, 学位授与番号 : 医博甲第1345号，学位授与年月日：平成11年3月31日,学位授与年：199

核内受容体PPAR-α遺伝子変異体機能解析と脂質代謝異常

金沢大学附属病院脂質代謝を制御している核内受容体の多型が高脂血症の遺伝的背景に与えている影響を明らかにするため以下の検討を行った.PCR-DGGE法および直接塩基配列決定法を用い高脂血症患者100例を対象とした検討から脂質代謝を調節する核内受容体であるPPARα遺伝子多型D140NおよびG395E変異,PPARγ2遺伝子P12A変異,PPARδ遺伝子ではThr411(silent C to T)多型を同定した.次にこれらの変異について,一般人口中での影響を検討した.一般人口男性298例中においてPPARα遺伝子G395Eを6例に,D140Nを2例に認めた.血清脂質値においてG395E保持者は有意に総コレステロール値およびLDLコレステロール値の上昇を認めたが,中性脂肪およびHDLコレステロール値の有意の変化は認めなかった.D140Nでは有意の血清脂質値の変化を認めなかった.PPARγ2遺伝子P12A変異は一般人の7%に存在し有意にHDL-C低値であった.PPARδ遺伝子ではThr411(Silent C to T)多型は特に血清脂質値などに影響は与えていなかった.ヒト肝癌細胞を由来とするHepG2細胞を用い,PPARαの発現ベクターと,その転写活性を評価するためのレポーターベクターをリポフェクション法にて細胞に導入した.レポーターベクターはホタルルシフェラーゼの上流にPPRE配列を挿入し,発現されたルシフェラーゼ量として測定できるようにしたものを用い,内部コントロールとしてウミシイタケルシフェラーゼを発現するベクターを用いることで,デュアルルシフェラーゼレポーターアッセイシステムとして発現量を測定した.結果in vitroの機能解析においてPPARα遺伝子G395EおよびD140N変異はいずれもwild typeと比較し有意に機能亢進を示した.高トリグリセライド血症においてPPARαを活性化するフィブラート剤を使用するとLDLコレステロール値が上昇する場合があるが,類似のメカニズムからPPARα機能亢進多型が高LDL血症をきたす遺伝素因となっている可能性が示唆された.研究課題/領域番号:13770643, 研究期間(年度):2001-2002出典：「核内受容体PPAR-α遺伝子変異体機能解析と脂質代謝異常」研究成果報告書　課題番号13770643（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-13770643/)を加工して作

代謝関連核内受容体遺伝子変異機能解析と肥満および脂質代謝異常

金沢大学医薬保健研究域医学系核内受容体は特有のリガンド存在下に多数の遺伝子群を協調的に発現制御する転写因子で,フィブラート剤チアゾリジン剤は,それぞれ核内受容体PPARαおよびγを標的としている.PPARsに限らずRetinoid-X-receptor (RXR)とヘテロダイマーを形成して作用する核内受容体は,近年代謝に重要な多数の遺伝子の制御が示され,MetSに代表される代謝疾患治療戦略の重要な標的となっている.しかし,動物実験では明らかでもヒトでも同じとは限らない.この点で機能的遺伝子変異の同定は重要な知見を与えるものであり,また核内受容体を標的とする治療の開発には必須の情報でもある.我々は高脂血症患者を対象としてPPARα,γ,δ,LXRα,Farnesoid-X-receptor (FXR),RXRγ遺伝子を検索,これまでに4個のミスセンス型(PPARαG395E, PPARαD140N, PPARγ2 P12A, RXRγG14S)と5\u27端非翻訳領域の多型1個(FXR -1g->t)を同定した.RXRγ S14多型保持者は一般人(5%)と比較しFCHLに有意に高頻度(15%)で,TG高値およびHDL-C低値であり,冠動脈造影施行105例でcoronary stenosis indexが有意に高値であった.機能解析でRXRγS14は機能亢進型であり,リポ蛋白リパーゼプロモーター配列の転写をより強く抑制した.これらの結果よりRXRγ遺伝子はFCHLの遺伝子背景に何らかの寄与をしていると考えられた.FXR遺伝子-1g->t多型では-1t保持者は有意にBMI高値,TC低値,拡張期血圧低値,腹部USで脂肪肝は低頻度と多彩な影響がみられた.またスタチンを投与した症例の解析では,FXR-1t保持者は-1gg保持者に比べより強くTchが低下,治療効果の予測にも重要と考えられた.研究課題/領域番号:17790603, 研究期間(年度):2005 – 2006出典：「代謝関連核内受容体遺伝子変異機能解析と肥満および脂質代謝異常」研究成果報告書　課題番号17790603（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-17790603/)を加工して作

Comparison of the effects of losartan vs. ramipril on several adipocytokines and vascular remodeling biomarkers

金沢大学医学系研究

Severe Renal Hemorrhage in a Pregnant Woman Complicated with Antiphospholipid Syndrome: A Case Report

Antiphospholipid syndrome is a systemic autoimmune disease with thrombotic tendency. Consensus guidelines for pregnancy with antiphospholipid syndrome recommend low-dose aspirin combined with unfractionated or low-molecular-weight heparin because antiphospholipid syndrome causes habitual abortion. We report a 36-year-old pregnant woman diagnosed with antiphospholipid syndrome receiving anticoagulation treatment. The patient developed left abdominal pain and gross hematuria at week 20 of pregnancy. An initial diagnosis of left ureteral calculus was made. Subsequently abdominal-pelvic computed tomography was required for diagnosis because of the appearance of severe contralateral pain. Computed tomography revealed serious renal hemorrhage, and ureteral stent placement and pain control by patient-controlled analgesia were required. After treatment, continuance of pregnancy was possible and vaginal delivery was performed safely. This is the first case report of serious renal hemorrhage in a pregnant woman with antiphospholipid syndrome receiving anticoagulation treatment and is an instructive case for urological and obstetrical practitioners

Serum lipoprotein lipase mass: Clinical significance of its measurement

金沢大学大学院医学系研究科Lipoprotein lipase (LPL) is a lipolytic enzyme involved in catalyzing hydrolysis of triglycerides (TG) in chylomicrons and very low-density lipoprotein (VLDL) particles. Over the last decade, increasing attention has been paid to the clinical significance of measuring serum LPL protein mass without heparin injection to the study subjects. In earlier studies, this marker was utilized to classify LPL deficient subjects, which is an extremely rare metabolic disorder with a frequency of one in one million. Later, researchers paid more attention to the clinical significance of measuring this parameter in more common metabolic disorders. Studies have shown that pre-heparin plasma or serum LPL mass has significant relationships with serum lipids and lipoproteins, visceral fat area, insulin resistance, and even the development of coronary atherosclerosis in cross-sectional studies, although this might be a metabolic surrogate marker with almost no catalytic activities, which does not appear to be involved in catalyzing hydrolysis of TG in TG-rich lipoproteins. Recently, a prospective study has demonstrated that low serum LPL concentration predicts future coronary events. Taken together, we suggest that pre-heparin LPL mass in plasma or sera provide us with useful and important information on the development of metabolic disorders leading to atherosclerotic disease. © 2006 Elsevier B.V. All rights reserved

Efficacy of colestimide coadministered with atorvastatin in Japanese patients with heterozygous familial hypercholesterolemia (FH)

金沢大学大学院医学系研究科　Background: Colestimide, a 2-methylimidazole-epichlorohydrin polymer, is a new bile-acid-sequestering resin, that is 4-fold as powerful at lowering low-density lipoprotein cholesterol (LDL-C) as the conventional resin (cholestyramine). Moreover, colestimide has excellent patient compliance because it is available in tablet form. Methods and Results: The clinical efficacy of colestimide coadministered with atorvastatin on lipid and apolipoprotein concentrations was examined in 15 patients (M/F = 10/5, mean±SE age=54±9 years) with heterozygous familial hypercholesterolemia (FH). After a period of wash-out of any lipid-lowering drugs, atorvastatin (20-40mg) was administered to patients for at least 8 weeks, and then 3 g of colestimide was administered for a further 8 weeks. Total and LDL-C significantly (<0.0001) decreased by 35% from 361 to 233mg/dl and 41% from 274 to 161 mg/dl, respectively. Addition of colestimide caused a further significant 12% and 20% reduction, respectively, from the initial values to 205 and 129 mg/dl, respectively. Colestimide was also effective in reducing serum LDL-C concentrations in heterozygous FH patients with hypertriglyceridemia (triglycerides ≥150mg/dl). Conclusions: When monotherapy with atorvastatin is insufficient to treat severely hypercholesterolemic patients, such as those with heterozygous FH, colestimide acts to reinforce the action of statins

Severe renal hemorrhage in a pregnant woman complicated with antiphospholipid syndrome: A case report

金沢大学医薬保健研究域医学系Antiphospholipid syndrome is a systemic autoimmune disease with thrombotic tendency. Consensus guidelines for pregnancy with antiphospholipid syndrome recommend low-dose aspirin combined with unfractionated or low-molecular-weight heparin because antiphospholipid syndrome causes habitual abortion. We report a 36-year-old pregnant woman diagnosed with antiphospholipid syndrome receiving anticoagulation treatment. The patient developed left abdominal pain and gross hematuria at week 20 of pregnancy. An initial diagnosis of left ureteral calculus was made. Subsequently abdominal-pelvic computed tomography was required for diagnosis because of the appearance of severe contralateral pain. Computed tomography revealed serious renal hemorrhage, and ureteral stent placement and pain control by patient-controlled analgesia were required. After treatment, continuance of pregnancy was possible and vaginal delivery was performed safely. This is the first case report of serious renal hemorrhage in a pregnant woman with antiphospholipid syndrome receiving anticoagulation treatment and is an instructive case for urological and obstetrical practitioners. Copyright © 2011 Shohei Kawaguchi et al

Chronological urodynamic evaluation of changing bladder and urethral functions after robot-assisted radical prostatectomy

Objective To examine chronological changes in urethral and bladder functions before, immediately after, and 1 year after robot-assisted radical prostatectomy (RARP), urodynamic studies were prospectively performed. Methods Sixty-three consecutive patients underwent pressure-flow studies, urethral pressure profiles, and abdominal leak point pressure (ALPP) tests 1-2 days before, immediately after, and 1 year after RARP. Results The mean bladder compliance was 28.3 mL/cm H2O before RARP; it worsened to 16.3 mL/cm H2O immediately after RARP and recovered to 27.1 mL/cm H2O at 1 year. The mean detrusor pressure at maximum flow rate was 61.9 cm H2O before RARP; it decreased to 34.3 cm H2O immediately after RARP and remained at 35.6 cm H2O at 1 year. The mean maximum urethral closure pressure was 84.2 cm H2O before RARP; it decreased to 33.4 cm H2O immediately after RARP and recovered to 63.0 cm H2O at 1 year. Intrinsic sphincter deficiency (ISD) evaluated by the ALPP test was observed in 53 patients immediately after RARP, although no patient showed ISD before RARP. ISD remained in 7 patients at 1 year. Both ALPP and maximum urethral closure pressure at 1 year were significant factors for continence in multivariate analysis. Conclusion Urethral sphincter and bladder function worsen immediately after RARP and recover over time. The bladder storage function after RARP returns to almost the same level before RARP, and the voiding function improves compared with the condition before RARP; however, the urethral sphincter function does not return to its preoperative level. Urethral sphincter dysfunction is considered the main factor for urinary incontinence after RARP. © 2015 Elsevier Inc.Embargo Period 12 month

Decreased ADP-Ribosyl Cyclase Activity in Peripheral Blood Mononuclear Cells from Diabetic Patients with Nephropathy

Aims/hypothesis. ADP-ribosyl-cyclase activity (ADPRCA) of CD38 and other ectoenzymes mainly generate cyclic adenosine 5’diphosphate-(ADP-) ribose (cADPR) as a second messenger in various mammalian cells, including pancreatic beta cells and peripheral blood mononuclear cells (PBMCs). Since PBMCs contribute to the pathogenesis of diabetic nephropathy, ADPRCA of PBMCs could serve as a clinical prognostic marker for diabetic nephropathy. This study aimed to investigate the connection between ADPRCA in PBMCs and diabetic complications. Methods. PBMCs from 60 diabetic patients (10 for type 1 and 50 for type 2) and 15 nondiabetic controls were fluorometrically measured for ADPRCA based on the conversion of nicotinamide guanine dinucleotide (NGD+) into cyclic GDP-ribose. Results. ADPRCA negatively correlated with the level of HbA1c (P = .040, R2 = .073), although ADPRCA showed no significant correlation with gender, age, BMI, blood pressure, level of fasting plasma glucose and lipid levels, as well as type, duration, or medication of diabetes. Interestingly, patients with nephropathy, but not other complications, presented significantly lower ADPRCA than those without nephropathy (P = .0198) and diabetes (P = .0332). ANCOVA analysis adjusted for HbA1c showed no significant correlation between ADPRCA and nephropathy. However, logistic regression analyses revealed that determinants for nephropathy were systolic blood pressure and ADPRCA, not HbA1c. Conclusion/interpretation. Decreased ADPRCA significantly correlated with diabetic nephropathy. ADPRCA in PBMCs would be an important marker associated with diabetic nephropathy