高血圧症における局所アンジオテンシンIIの冠動脈狭窄に果たす役割

金沢大学医学部附属病院1.高血圧の実験モデル動物とされる高血圧自然発症ラット(SHR)を用い血漿エンドセリン(ETー1)濃度および腸間膜動脈から放出されるETー1を測定し、対照ラットと比較した。その結果血漿ETー1濃度はSHRおよび対照ラットとも加齢と共に増加したがいずれの週齢に於てもSHRでは対照に比べ高値を呈した。特に高血圧の発症前(5週齢)において血漿ETー1濃度がSHRで有意に高値(20vs 13.8pg/ml)であったことは高血圧の維持のみならずその発症機構にも当該ペプチドの関与が示唆される。腸間膜灌流実験系に於てもSHRでETー1の放出量は多く血漿濃度を反映する結果であった。2.血管障害を来たしやすい糖尿病の実験モデル動物とされるストレプトゾトシン糖尿病ラット(STZーDM)においても同様の検討を行なった。その結果STZーDMでは対照ラットに比べ血漿ETー1濃度5.1vs3.0pg/mlP〈0.05)および腸間膜灌流液中のETー1量(35.8vs14.9pg/ml/h)は有意に高値であった。このことはSTZーDMにおいては血管内皮の障害が生ずる結果、ET産生が亢進する事を示しており、血漿ETー1濃度が血管障害のマ-カ-となる可能性を示唆する成績である。3.インタ-ロイキン(IL)はサイトカインの一種として血管内皮細胞においても細胞機能の調節に関与する可能性がある。腸間膜灌流実験系での検討では、ILー2がETの産生亢進することが示された。ILー2は抗腫瘍薬として臨床応用が期待されているが、血管内皮への影響を知るマ-カ-としてETの血漿濃度の測定が有用である。4.以上の結果をまとめると血管内皮および血管平滑筋細胞では局所的に産生されるペプチドが血管の緊張や他の機能を調節しており一部血管障害の病因にも関与する事が明かとなった。今後、ETの受容体拮抗剤を用いることにより、治療への展望に関しても実験を重ねたい。In spontaneously hypertensive rats, plasma endothelin concentration as well as the peptide released from the mesenteric arteries were significantly increased both at 5 and 10 weeks of age. This finding suggests that endothelin may contribute for the development of hypertension in this, model. We also studied the vascular endothelin production in the streptozotocin induced diabetic rats. In diabetic rats, endothelin was significantly increased as compared with the control rats indicating that endothelin may serve as a marker for the vsacular insults in diabetes associated with microangiopathy. We assume that endothelin was increased in diabetic rats as a result of functional and/of structural derangement caused by streptozotocin diabetes. In the last part of the experiment, we measured plasma and arterial endothelin in rats treated with interleukin 2, an agent reported to cause extravasation of fluid in the third space, presumably as a result of endothelial damage. We conclude that plasma endothelin will be used as a marker for the endothelial function in this condition. We plan to undertake further study as to the possibility of endothelin receptor antagonist for the therapeutic purpose.研究課題/領域番号:02454219, 研究期間(年度):1990 – 1991出典：研究課題「高血圧症における局所アンジオテンシンIIの冠動脈狭窄に果たす役割」課題番号02454219（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-02454219/024542191991kenkyu_seika_hokoku_gaiyo/）を加工して作

血管障害の発生および進展に果たす血管壁レニン・アンジオテンシンの役割り

金沢大学医学部附属病院流血中に存在するレニン及びアンジオテンシンは従来血圧調節に重要な循環因子として数多くの研究がなされてきた。近年、血管壁、副腎、精腺などの局所組織中にもレニン・アンジオテンシン系の各コンポネントが存在することが明らかになり、その病態生理的意義について解明されつつある。我々は、血管壁で産生されるアンジオテンシンIIの血流の調節や、血管障害に関与する可能性に注目し、主として動物実験を行なった。又、関連する血管作動性物質として、近年、血管内皮細胞で産生されることが明らかとなったエンドテリンについてもその局所血管作用について検討した。更に副腎ステロイドが内皮細胞に特異的受容体を有することから、21ーデオキシアルドステロンの作用についてもin vivo,in vitroの両面から検討した。その結果、動脈壁からは腸間脈動脈から、アンジオテンシンIIが産生され、病態の変化によりその産生量は異なることが明かとなった。脳血管障害の発生及び進展にこれらの生理活性物質がどの程度関与するかは依然検討すべき課題であるが、その可能性は否定できないと考えられる。Angiotensin II (ANG II) generation from the isolated mesenteric arteries was measured in control rats, and in hypertensive rat models. In order to elucidate the possible role of the vascular renin-angiotensin system (RAS) for the control of blood pressure, the effect of angiotensin converting enzyme (ACEI) on the ANG II production was also examined. The perfusion pressure (PP) response to endothelin (ET), a novel vasoconstrictor peptide, was also examined in the mesenteric artery preparation obtained from rats with various ANG II levels. In the control rats, 43.012.0 pg/h of ANG II was released in the perfusate, which was not significantly changed by nephrectomy (Nex), in spite of the decreased plasma renin activity (PRA) and plasma ANG II concentration in the circulation. ACEI treatment significantly decreased blood pressure (BP) in the control and Nex rats, in parallel with a decreased vascular ANG II generation. In deoxycorticosterone acetate-treated rats, PRA, plasma aidosterone and ANG II concentration were significantly suppressed, and ACEI administration induced a slight but significant decrease in BP. ET produced a sustained increase in PP in the control rat mesenteric arteries. The increase was more potent than ANG II on molar basis, and was not inhibited by ANG II analogue. In conclusion, the present results support that the ANG II is generated in the vascular wall independent of RAS in the circulation, and suggest that the vascular RAS is partly responsible for the antihypertensive action of ACEI. A lack of inhibition of pressure response to ET by ANG II analogue suggests that ET and ANG II possess their own specific receptor研究課題/領域番号:63440091, 研究期間(年度):1988 – 1989出典：研究課題「血管障害の発生および進展に果たす血管壁レニン・アンジオテンシンの役割り」課題番号63440091（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-63440091/634400911989kenkyu_seika_hokoku_gaiyo/）を加工して作

血管内皮細胞におけるアルドステロンの産生とその調節因子に関する研究

金沢大学医学部最近の分子生物学的手法によりレニン・アンジオテンシン(R-A)系を構成する各要素は腎、副腎、性腺系に存在することが明らかになり、循環血中R-A系と独立して作動していると考えられている。現在、心血管系に局在するR-A系についても研究が進められているが産生経路、調節因子、生理的意義の詳細は不明である。本研究で我々は、血管壁がR-A系の最終活性物質であるアルドステロンを産生することを見いだした。先ず初年度ではラット腸間膜動脈かん流実験からアルドステロンがガスクロマトグラフィー質量分析器(GC-MS、Schimaz QP1000、既存)により検出されることを報告した(Hypertension誌 1995 in press)。最終年度ではアルドステロン合成酵素(P450aldo)の発現実験を行い、分子生物学的手法を用いて局所における産生を証明した。即ち、培養ヒト肺動脈由来内皮細胞および平滑筋細胞からPoly(A)+mRNAを抽出し調製液、primer,Reverse Transcriptase添加後42℃60分でインキュベート。99℃、5分間でsingle strand cDNAを得た後2.5unitのTaq DNA polymeraseで増幅した。RT-PCRによって得られるproductのサイズは322bpであり、1.5%agarose gelで電気泳動しナイロン膜に写した。膜はSSC、SDSで洗浄、オートラジオグラムにかけ、イメージアナライザーで解析した。一方、培養液中の産生ステロイドの微量分析はHPLC(ファルマシア、既存),GC-MSを用いて分析した。その結果、副腎外組織である血管壁においてアルドステロンが産生されることが確認され、その調節因子についても新たな知見が得られた(Hatakeyama et al.JBC269:24316,1994)。その産生量は副腎に比べて1:50と極めて少ないが、autocrine/paracrineとして血管局所において生理作用を発揮していると考えられた。特に、アルドステロンがアンジオテンシンIIの血管平滑筋肥大作用に対して促進的に作用するという今回の結果は、世界に先駆けた概念でありVascular aldosterone systemと命名された。In the current study, we identified that aldosterone, a potent mineralocorticoid which is synthesized exclusively in the adrenal cortex, is also produced in the vascularture. In the first year, we undertook a perfusion experiment of the rat mesenteric arteries and identified an aldosterone like immunoreactivity in the HPLCseparated perfusate. This fraction was further analyzed by GCMS which confirmed that immunoreactive peak was identical to aldosterone molecule. In the second year, we performed gentic expression of P450aldo (aldosteone synthetase) mRNA in human endothelical cells. P450aldo mRNA was detected at a concentration of 1/50 compared with the adrenal gland. We hypothesized that vascular aldosterone may contribute to the vascular remodelling and to the pathogensis of hypertension in autocrine/paracrine fashion研究課題/領域番号:05454318, 研究期間(年度):1993 – 1994出典：研究課題「血管内皮細胞におけるアルドステロンの産生とその調節因子に関する研究」課題番号05454318（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-05454318/054543181994kenkyu_seika_hokoku_gaiyo/）を加工して作

プロスタサイクリン(PGI_2)の分泌調節に関する研究

金沢大学医学部研究課題/領域番号:58570995, 研究期間(年度):1983出典：研究課題「プロスタサイクリン(PGI_2)の分泌調節に関する研究」課題番号58570995（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-58570995/）を加工して作

A proximal direct repeat motif characterized as a negative regulatory element in the human renin gene

The regulation of renin gene expression is thought to be fundamental to regulation of the total renin-angiotensin system. The human renin gene contains a direct repeat (DR) motifAGGGGTCAC-AGGGCCA in the proximal region (-259/-245 bp), which contains similar sequence for nuclear receptor superfamily binding core motif, AGGTCA, and is the most similar to COUP-TFII consensus. The DR motif was evaluated as a functional cis-element with renal cortex and chorio-decidual cells by footprint assay, electromobility shift assay (EMSA) and reporter assay. The DR motif site was protected by footprint analysis with a clear hypersensitive and a minor hypersensitive region in good accordance with the DR of the consensus. One of the binding proteins was strongly suspected to be COUP-TFII-consensus-specific by EMSA. The DNA/protein complexes obtained with nuclear extract of renin producing cells could be completely blocked by homologous competitor and strongly blocked by the second-half mutant oligonucleotide of the DR motif but not by the first-half mutant oligonucleotide. Finally, the transcriptional activity of second-half mutant construct is slightly elevated and that first-half mutant construct is significantly stronger by twofold compared with wild type construct in reporter assay. These findings suggest that the DR motif site of the human renin gene functions as a negative regulatory element involved in a twofold repression of transcription and that member(s) of nucleic receptor superfamily bind the site and play important roles in the human renin gene expression with a possibility that one of the binding protein is COUP-TFII

A case report of primary sclerosing cholangitis with ulcerative colitis

金沢大学大学院医学系研究科がん細胞学A 24-year-old women was admitted to our hospital for evaluation of hepatic dysfunction. She was diagnosed as ulcerative colitis in our hospital in 1987 (at age of 16). She was treated with salazosulfapyridine and prednisolone, and she had been well with no relapse for four years. Endoscopic retrograde cholangiogram showed beaded appearances of the intrahepatic bile ducts and irregular stricture of the intra- and extrahepatic bile ducts. Hepatic needle biopsy revealed mild periductal fibrosis. She had no history of bile duct surgery and choledochlithiasis. From overall results, she was diagnosed as primary sclerosing cholangitis. We confirmed that the remission of ulcerative colitis by barium enema study and colonoscopy. Reported cases of primary sclerosing cholangitis associated with ulcerative colitis in Japan were reviewed. The most cases had pancolitis, and sclerosing change of intra- and extrahepatic bile ducts. Whether primary sclerosing cholangitis might precede or follow ulcerative colitis remains to be controversial. This case was daignosed before the development of irreversible fibrosis, and responsive to steroid therapy

Cromakalim, a vasodilator, differentially inhibits Ca2+ currents in NG108-15 neuroblastoma × glioma hybrid cells

AbstractExtracellular perfusion with the antihypertensive agent cromakalim produced an inhibition of 22–66% in the low-threshold transient Ca2+ (T-like) current in NG108-15 hybrid cells. Cromakalim suppressed the high-threshold and long-lasting Ba2+ current (L-like Ca2+ current) by 29–73%, but had almost no effect on the high-threshold and inactivating Ba2+ current (N-like Ca2+ current). IC50 for T-like and L-like currents was the same at about 100 μM. The inhibitory effect developed relatively fast and was reversible. These results indicate that cromakalim can selectively inhibit the activity of inward Ca2+ currents

カブトプリル投与中の分腎機能変化-腎血管性高血圧における診断的意義-

金沢大学大学院医学系研究

Changes in aromatase (CYP19) gene promoter usage in non-small cell lung cancer

金沢大学医薬保健研究域医学系In humans, aromatase (CYP19) gene expression is regulated via alternative promoters. Activation of each promoter gives rise to a CYP19 mRNA species with a unique 5′-untranslated region. Inhibition of aromatase has been reported to downregulate lung tumor growth. The genetic basis for CYP19 gene expression and aromatase activity in lung cancer remains poorly understood. We analyzed tissues from 15 patients with non-small cell lung cancer (NSCLC) to evaluate CYP19 promoter usage and promoter-specific aromatase mRNA levels in NSCLC tumor tissues and adjacent non-malignant tissues. CYP19 promoter usage was determined by multiplex RT-PCR and aromatase mRNA levels were measured with real-time RT-PCR. In non-malignant tissues, aromatase mRNA was primarily derived from activation of CYP19 promoter I.4. Although promoter I.4 usage was also dominant in tumor tissues, I.4 activation was significantly lower compared with adjacent non-malignant tissues. Activity of promoters I.3, I.1 and I.7 was significantly higher in tumor tissues compared with non-malignant tissues. In 4 of 15 cases of non-small cell lung cancer, switching from CYP19 promoter I.4 to the alternative promoters II, I.1 or I.7 was observed. In 9 cases, there were significantly higher levels of aromatase mRNA in lung tumor tissues compared with adjacent non-malignant tissues. These findings suggest aberrant activation of alternative CYP19 promoters that may lead to upregulation of local aromatase expression in some cases of NSCLC. Further studies are needed to examine the impact of alternative CYP19 promoter usage on local estrogen levels and lung tumor growth. © 2011 Elsevier Ireland Ltd. All rights reserved

Dual-time-point 18F-FDG PET imaging for diagnosis of disease type and disease activity in patients with idiopathic interstitial pneumonia

Purpose Individual clinical courses of idiopathic interstitial pneumonia (IIP) are variable and difficult to predict because the pathology and disease activity are contingent, and chest computed tomography (CT) provides little information about disease activity. In this study, we applied dual-time-point [18F]-fluoro-2-deoxy-D-glucose (18F-FDG) positron emission tomography (PET), commonly used for diagnosis of malignant tumors, to the differential diagnosis and prediction of disease progression in IIP patients. Methods Fifty patients with IIP, including idiopathic pulmonary fibrosis (IPF, n=21), nonspecific interstitial pneumonia (NSIP, n=18), and cryptogenic organizing pneumonia (COP, n=11), underwent 18F-FDG PET examinations at two time points: Scan 1 at 60 min (early imaging) and Scan 2 at 180 min (delayed imaging) after 18F-FDG injection. The standardized uptake values (SUV) at the two points and the retention index (RI-SUV) calculated from them were evaluated and compared with chest CT findings, disease progression, and disease types. To evaluate short term disease progression, all patients were examined pulmonary function test every 3 months for 1 year after 18F-FDG PET scanning. Results The early SUV for COP (2.47±0.74) was significantly higher than that for IPF (0.99±0.29, P=0.0002) or NSIP (1.22±0.44, P=0.0025). When an early SUV cut-off value of 1.5 and greater was used to distinguish COP from IPF and NSIP, the sensitivity, specificity, and accuracy were 90.9%, 94.3%, and 93.5%, respectively. The RI-SUV for IPF and NSIP lesions was significantly greater in patients with deteriorated pulmonary function after 1-year of follow-up (progressive group, 13.0±8.9%) than in cases without deterioration during the 1-year observation period (stable group, -16.8±5.9%, P<0.0001). However, the early SUV for all IIP types provided no additional information of disease progression. When an RI-SUV cut-off value of 0% and greater was used to distinguish progressive IIPs from stable IIPs, the sensitivity, specificity, and accuracy were 95.5%, 100%, and 97.8%, respectively. Conclusion Early-SUV and RI-SUV obtained from dual-time point 18F-FDG PET are useful parameter for the differential diagnosis and prediction of disease progression in patients with IIP