一氧化氮在梗阻性黄疸大鼠肝、肾、肠组织中含量变化及意义

目的 了解一氧化氮（NO）在梗阻性黄疸大鼠肝，肾，肠组织中含量变化及意义。方法 大鼠胆总管结扎后，分别于第一周内和第三周内变化Aminoguanidine(AG)抑制NO合成，同时应用生理盐水（NS）作对照，检测不同时段抑制NO合成后大鼠肝，肾，肠组织中NO和丙二醛（MDA）含量，肌酐清除率（Ccr)，血清总胆红素（T－BIL）和丙氨酸氨基转移酶（ALT）含量及肠系膜淋巴结细菌移位（BT）率的变化。结果 胆总管结扎后，大鼠肝，肾，肠组织中NO含量明显升高，在胆总管结扎第一周抑制NO合成后，肝，肾肠组织中NO含量明显下降，MDA含量明显升高，血ALT明显升高，Ccr明显下降，肠系膜淋巴结BT率明显升高；而在胆总管结扎第三周抑制NO合成后，肝，肾，肠组织中NO和MDA含量明显下降，血ALT明显下降，Ccr明显升高，肠系膜淋巴结BT率明显下降。结论 NO在胆道梗阻引起的肝，肾，肠粘膜屏蔽功能障碍的发生机制中具有重要作用，既有保护作用，又有损害作用。梗阻早期表现为对组织的保护作用，后期表现为对组织的损害作用。     

梗阻性黄疸病人麻醉手术期间一氧化氮的变化

梗阻性黄疸病人麻醉手术期间一氧化氮的变化黄秀芳金烈烈陈必成１９８６年确定一氧化氮（ＮＯ）为血管内皮衍生松弛因子后，很快就认识到它在机体内有广泛的生理病理作用。连续硬膜外阻滞常致血压和心率降低，梗阻性黄疸病人的变化更为显著，本文旨在观察梗阻性黄疸病人麻...     

梗阻性黄疸大鼠血浆和肾组织内皮素含量的变化及与肾功能关系的研究

目的：了解血浆，肾组织内皮素（ET）在梗阻性黄疸（OJ）大鼠模型上不同梗阻时间的变化情况，探讨循环ET和肾内ET与梗阻性黄疸时肾功能损害的关系。方法：胆总管结扎及假手术组各32只Wistar大鼠分别于术后5，10，15，20d（每小时n=8)采用放射免疫法测定血浆和肾皮质，髓质细胞匀浆ET含量，同时测定血清肌酐（Cr)，尿素氮（BUN）和直接胆红素（DB）浓度，并对肾脏行光镜下病理形态学观察。结果；血浆ET浓度及肾皮质，髓质ET含量在梗阻5d即升高，且随OJ时间延长进一步升高，伴有血清Cr,BUN,DB的升高和肾脏病毒形态的进行性改变。血浆ET，肾皮质ET在大部分或部分时相与Cr,BUN呈明显正相关，血浆ET在大部分时相与DB呈明显正相关。结论：梗阻性黄疸时循环和肾内ET水平持续性升高；血浆和肾皮质ET的升高可能参与了肾脏滤过功能的损害；肾髓质ET的升高可能是导致OJ早期尿浓缩功能异常的介质之一；高胆红素血症可能是血浆ET升高的原因之一。     

梗阻性黄疸大鼠小肠肠系膜微循环变化及意义

目的以肠系膜为微循环观测窗，探讨梗阻性黄疸（简称梗黄）对肠系膜微循环的影响。方法将60只SD大鼠均分为对照组和梗黄组，每组分1、3、7d三个观测时相。将大鼠腹腔内一段回肠袢肠系膜平铺于恒温灌流盒作为微循环观察窗，用微循环显微镜及视频图像分析系统观测肠系膜微循环的动态变化并记录结果。结果梗黄组大鼠肠系膜微循环从微血管形态、微血管流态及微血管周围状态三个方面均发生明显变化，主要表现为毛细血管与微静脉扩张，血流减慢，红细胞聚集，微栓形成、毛细血管通透性增加及出血等，与对照组各相同时相比较差异均有统计学意义（P〈0．05）。结论梗阻性黄疸可引起大鼠肠系膜微循环障碍。     

肿瘤坏死因子、内皮素-1和一氧化氮在梗阻性黄疸大鼠血浆和肝组织中活性变化及甘氨酸治疗的研究

梗阻性黄疸 (以下简称梗黄 )时存在明显的内毒素血症[1 ] ,肿瘤坏死因子 (ANF α)、内皮素 1(ET 1)和一氧化氮(NO)的产生与内毒素关系密切。有研究报道 ,甘氨酸能有效抑制内毒素的生物学效应[2 4 ] ,本项研究探讨梗黄时TNF α、ET 1和NO的变化及甘氨酸对它们的影响。材料与方法1.材料 :同龄标准Wistar大鼠 90只 ,雌雄各半 ,体重2 5 0～ 30 0 g。鲎试剂盒由上海伊华医学科技有限公司提供。肿瘤坏死因子放免药盒、内皮素放免药盒和一氧化氮代谢产物硝酸盐测定试剂盒由解放军总医院放免研究所提供。甘氨酸由上海生物制品研究所提供。2 …     

大鼠梗阻性黄疸形成中血及肝组织内皮素的水平及意义

目的　了解梗阻性黄疸 (obstructivejaundice ,OJ)形成过程中内皮素 (endothelin ,ET)在血浆及肝组织中的水平及其在OJ肝损伤中的作用。方法　以大鼠胆总管结扎 (bileductligation ,BDL)为模型 ,采用放射免疫法 (radioimmunassay ,RIA)测定了BDL后 3、6、9、1 2、1 5d时血浆及肝组织中ET的含量 ,同时观测血清谷丙氨酸转氨酶 (ALT)含量变化 ,并与肝组织病理改变进行对照分析。结果　BDL后血清总胆红素 (totalbilirubin ,TBi)迅速升高 ,明显高于NC组和SO组 (P <0 0 1 ) ;BDL组血浆及肝组织中ET水平明显高于对照组 (P <0 0 1 ) ,BDL各时相组差异有显著性意义 (F =1 8 41 ,P <0 0 1和F =40 61 ,P <0 0 1 ) ,随梗阻时间延长呈进行性含量升高 ,1 5dET值最高 ,分别为 (2 94 98± 35 95) pg/ml和 (2 0 1 86± 2 4 45) pg/ 1 0 0mg ;二者与血清ALT均呈正相关 (r =0 8441 ,P <0 0 1和r=0 930 3 ,P <0 0 1 ) ,病理切片见肝细胞从肿胀、空泡变性到灶状片状坏死 ,损伤也呈进行性加重 ,与ET含量变化一致 ;病理切片还显示肝内有大量的小胆管及毛细胆管的增生 ,且随着胆管梗阻时间的延长 ,增生越来越明显。结论　实验结果提示在OJ形成中血及肝组织中ET含量上升 ,并随梗阻时间延长而加重 ;ET可能是参与OJ肝脏     

梗阻性黄疸机体补体变化研究及意义

目的 探讨梗阻性黄疸患者补体功能变化。方法 观测１４例ＡＯＳＣ急诊手术患者（Ａ组），２３例择期手术梗阻性黄疸患者（Ｂ组）和１２０例胆囊结石患者（Ｃ组）围手术期Ｃ３、Ｃ４、ＣＩＣ和ＣＨ５０变化。资料经方差分析。结果 术前Ａ、Ｂ两组Ｃ３显著低于Ｃ组（Ｐ〈０．０１）。术后Ｂ组Ｃ３显著增加，Ａ组无明显变化，尤其ＣＩＣ仍处于较高水平。结论 梗阻性黄疸时Ｃ３产生和ＣＩＣ清除降低，可能与内毒素对枯否细胞损害和门     

内皮素在梗阻性黄疸肝、肾、心组织损伤中的作用及血塞通干预

目的 探讨内皮素（ET）在梗阻性黄疸肝、肾、心组织损伤中的作用机理及血塞通注射液的保护作用。方法 健康日本大耳白兔72只，雌雄各半，采用完全随机分组方法随机均分为3组：胆总管结扎（BDL）组、BDL＋血塞通注射液组及假手术组。建立实验动物模型，分别于术后3、6、9及21d取材。用全自动生化分析仪测定各组动物血清总胆红素、谷丙转氨酶、尿素氮及肌酐；用放免法测定各组动物肝、肾、心组织及血浆中的ET水平。结果 BDL组和BDL＋血塞通注射液组与假手术组比较，动物血浆及肝、肾、心组织中的ET含量明显升高（P〈0．01），肝、肾功能损害更严重（P〈0．01）；BDL＋血塞通注射液组比BDL组血浆及组织中ET含量明显降低（P〈0．05），肝、肾功能损害亦减轻（P〈0．05）。结论梗阻性黄疸可导致血浆及肝、肾、心组织ET含量升高，且随梗阻时间延长升高愈明显；血塞通注射液通过降低血浆及组织的ET水平，发挥对梗阻性黄疸肝、肾、心损害的保护作用。     

梗阻性黄疸肠外营养支持的重要意义

梗阻性黄疸肠外营养支持的重要意义同济医科大学器官移植研究所（４３００３０）刘敦贵，夏穗生梗阻性黄疸（以下简称梗黄）术前和术后能否施行肠外营养（Ｐａｒｅｎｔｅｒａｌｎｕｔｒｉｔｉｏｎ，ＰＮ），至今尚有争议。本文拟就使黄的发生机理、病生特点和ＰＮ支持的必...     

Increased nitric oxide production in hepatocytes is involved in liver dysfunction following obstructive jaundice

BACKGROUND: Obstructive jaundice damages critical functions in the liver. However, the mechanisms involved in hepatic dysfunction are obscure. Nitric oxide is implicated in liver injury under various pathological conditions. We previously reported that proinflammatory cytokine interleukin-1beta (IL-1beta) stimulated the production of nitric oxide in hepatocytes, which was associated with mitochondrial dysfunction. Studies were performed to examine whether obstructive jaundice influences the production of nitric oxide in hepatocytes and alters hepatic energy metabolism. MATERIAL AND METHODS: Hepatocytes were isolated and cultured from a rat model of obstructive jaundice or sham control. Nitric oxide production, ATP content, and ketone body ratio (acetoacetate/beta-hydroxybutyrate; KBR) were compared between the two groups in the presence of IL-1beta. RESULTS: Hepatocytes obtained from obstructive jaundice rats markedly increased the levels of nitric oxide production stimulated by IL-1beta compared with those from sham control. Western blot analysis revealed that the enhancement of nitric oxide production was a posttranslational event, since protein levels of inducible nitric oxide synthase (NOS) were unchanged between the two groups. IL-1beta decreased cellular ATP content in obstructive jaundice but not in sham control. Further, the KBR, which is a marker of mitochondrial redox state, was lower in obstructive jaundice than in sham control. Addition of N(G)-monomethyl-L-arginine, an inhibitor of NOS, abolished the decreases in ATP content and KBR as well as the nitric oxide production. CONCLUSIONS: These results indicate that a priming of nitric oxide production following obstructive jaundice is associated with the alteration of hepatic energy metabolism in part through mitochondrial dysfunction. Regulation of nitric oxide production may be a useful therapy for preventing liver dysfunction in obstructive jaundice.     

Expression of nitric oxide and inducible nitric oxide synthase in acute renal allograft rejection in the rat

BACKGROUND: Recent studies have shown that nitric oxide (NO) synthases, particularly inducible nitric oxide synthase (i-NOS), are induced in acute rejection episodes following heart, liver, pancreas and kidney allotransplantation. Furthermore, tissue and cellular injury has been demonstrated to be mediated by peroxynitrite (ONOO-), a metabolite of NO as well as a potent oxidant. However, a detailed relationship between NO, i-NOS and graft injury in transplantation remains elusive. METHODS: The present study used the following models of renal transplantation in rats: allografts (n = 5, Brown-Norway to Lewis [LEW] rats), isografts (n = 5, LEW to LEW) and allografts treated with aminoguanidine (AG), an i-NOS inhibitor (n = 5). Blood urea nitrogen (BUN), serum creatinine (SCr) and urinary and serum nitrosocompounds (NOx) were measured on days 2, 4 and 7 post-transplant. Western blot analysis of i-NOS protein expression and measurement of i-NOS activity were carried out in grafts harvested on Day 7, along with immunohistochemical and histopathological examinations. RESULTS: In the allograft group, both BUN and SCr levels increased markedly on Day 7, in parallel with a sharp increase in NOx. A band stained by anti-i-NOS antibody was detected at approximately 130 kDa, along with high levels of i-NOS activity and diffusely distributed i-NOS-positive cells (macrophages). Histologically, an acute rejection episode was confirmed (Grade 3 according to Banff classifications). In the AG group, reduced renal function and graft injury were significantly less severe than in the allograft group. CONCLUSIONS: In rat renal allograft acute rejection, markedly increased levels of serum NOx were observed, along with enhanced tissue i-NOS activity, together resulting in graft injury. AG administration suppressed the increase of serum NOx levels, with concomitant mitigation of tissue injury and renal function impairment.     

Role of inducible nitric oxide synthase in pig liver transplantation

BACKGROUND: Previously, we clarified the role of inducible nitric oxide synthase (iNOS) and the protective effect of an iNOS inhibitor in warm ischemia and reperfusion model. In this study, we investigated whether the same effects would be obtained by iNOS inhibitor in liver transplantation model. MATERIAL AND METHODS: Orthotopic liver transplantation was performed in pigs in the usual manner after about 6 h of cold preservation in University of Wisconsin solution. Aminoguanidine hemisulfate (AG) was used as the iNOS inhibitor and AG was administered intraportally at the dose of 10 mg/kg just after reperfusion. Two experimental groups were subjected, control group (n = 10), and AG group (n = 10). We investigated changes of serum nitrite/nitrate (NOx) and aspartate aminotransferase (AST). Expression of iNOS was examined by immunohistochemistry, including a double immunofluorescnce technique in combination with cofocal laser scanning microscopy. RESULTS: Serum NOx and AST were significantly lower in the AG group. Histological hepatic damage and thrombocyte thrombi were attenuated in the AG group. Expression of iNOS was recognized strongly at Kupffer cells and neutrophils in the centrilobular region of liver after reperfusion by cofocal laser scanning microscopy. Moreover, iNOS staining was attenuated in AG group compared with control group. CONCLUSIONS: These results indicate that hepatic ischemia and reperfusion injury in liver transplantation might be triggered by iNOS expression of Kupffer cells and neutrophils, and attenuated by administration of an iNOS inhibitor. Moreover, AG showed down regulation of iNOS expression after reperfusion.     

内毒素引起阻塞性黄疸大鼠肾功能障碍的机制

目的 探讨阻塞性黄疸(obstructive jaundice,OJ)时内毒素引起肾功能障碍的机制.方法 SD大鼠60只,胆总管结扎后,分5 d(B1),10 d(B2),15 d(B3)三组,每组各10只,同时建立相应对照组(A1,A2,A3),另30只胆总管结扎后分3组(SHUD,LAC,NS),每组各10只,分别用2 ml舒胆合剂、乳果糖液、生理盐水灌胃,连用9 d.观察内毒素、血和肾组织中内皮素(endothelin,ET)、一氧化氮(nitric oxide,NO)的含量、一氧化氮合酶(nitric oxide synthase,NOS)活性及肝、肾功能的变化.结果 血内毒素与血、肾组织ET含量,ET/NO比值呈显著正相关(P＜0.05,r=0.630,0.438,0.496,0.453),与肌肝清除率(creatinine clearance,Ccr)和肾皮质血流量(renal cortical blood flow,BCBF)呈显著负相关(P＜0.05,r=-0.600,-0.410).血、肾组织ET/NO比值与Ccr,RCBF呈显著负相关(P＜0.05,r=-0.449,-0.558,-0.626,-0.731).血和肾组织内NO水平与内毒素水平呈负相关(P＜0.05,r=-0.518,-0.441),与Ccr、RCBF呈正相关(P＜0.05,r=0.422,0.496,0.400,0.659).SHUD组与LAC血内毒素、ET水平组明显降低,血和肾组织NO,NOS活性以及Ccr,RCBF较NS组明显升高.结论 OJ时内毒素可通过刺激ET的释放,提高ET/NO比值,使肾内缩血管因子与扩血管因子比例失调而损伤肾功能.     

原发性肝癌合并阻塞性黄疸的诊断和治疗

目的 探讨原发性肝癌合并阻塞性黄疸的诊断与治疗。方法 自1996年10月至2002年12月间我院收治原发性肝癌合并阻塞性黄疸70例,分成积极治疗组(n=61)和对症处理组(n=9);积极治疗组再分为单纯胆道引流组(n=43)和综合治疗组(n=18)。积极治疗组接受胆道引流、手术和肝动脉化疗栓塞等治疗。随访所有病人的生存时间。结果 随访50例,随访率71.4％。积极治疗组和对症处理组平均生存时间分别为9.6个月和2.3个月,相差显著(t’=3.45,P<0.05)。综合治疗组和单纯胆道引流组平均生存时间分别为21.9个月和4.1个月,相差显著(t’=4.11,P<0.05)。结论 原发性肝癌合并阻塞性黄疸的病人如能及时解除黄疸,积极治疗肿瘤,仍可显著延长病人生命。     

Changes in nitric oxide production in the rat kidney due to CaOx nephrolithiasis

AIMS: The aim of the study was to test the hypothesis that the renal nitric oxide (NO) system is involved in the animal model of nephrolithiasis by evaluating the relationship between nitric oxide synthase (NOS) and oxidative stress. METHODS: Deposition of renal calculi was induced by adding 0.75% ethylene glycol (EG) to the drinking water of male Wistar rats. After 42 days of treatment, urinary biochemistry and urinary levels of oxalate, NO metabolites (nitrate and nitrite), cGMP, and lipid peroxides, and markers for renal damage and oxidative stress in the kidney were examined. In the second part of the experiment, two diuretic stimuli (intrarenal infusion of l-arginine or saline loading) were applied to test the renal NO system response. Finally, levels of three isoforms of NOS in renal tissues were evaluated by immunostaining. RESULTS: In the EG-treated rats, increased urinary excretion of enzymes and lipid peroxides and increased nitrotyrosine levels and oxidative injury markers in the kidneys indicated that peroxynitrite formation occurred during oxidative stress, while the 24-hr urinary excretion of NO metabolites and cGMP remained unchanged. In contrast to control rats, urinary excretion and NO metabolites and cGMP excretion were unresponsive to intrarenal l-arginine infusion; in response to saline loading, an increase in these factors was seen, but the increase was only 50% of that seen in the identically treated control group. A significant decrease in eNOS expression and increase in iNOS expression were observed in the renal medulla of the EG-treated group, whereas expression of nNOS was not affected. CONCLUSIONS: Although basal renal NO production remained unchanged, excessive peroxynitrite formation in the kidney was noted in this model. A decreased response of the NOS system was noted when diuretic stimuli were applied. How the imbalance between eNOS and iNOS expression influences CaOx stone formation requires detailed evaluation.     

N-乙酰半胱氨酸对大鼠胆道梗阻肝功能损伤的保护作用及机制研究

目的 探讨N-乙酰半胱氨酸(N-acetylcysteine,NAC)对大鼠胆道梗阻所致肝功能损伤的保护作用及其机制。方法Wistar大鼠72只随机均分成3组：(1)胆道结扎+NAC组(DBL+NAC,n=24)：开腹结扎并切断胆总管,建模成功后经腹腔注射NAC（150 mg·kg-1·d-1）连续注射7 d;(2)胆道结扎组（DBL组,n=24）;(3)假手术组(SO组,n=24)：仅行开腹游离胆总管不予结扎和切断。建模成功后1、3、5、7d每组分别活杀6只大鼠,取静脉血及肝组织,检测肝功能、血浆肿瘤坏死因子α(TNF-α)在各时相点的变化并采用Griess法检测一氧化氮(NO)产生情况。结果 在DBL组、DBL+ NAC组谷-草转氨酶(AST)、血清谷丙转氨酶(ALT)、总胆红素(TBIL)、直接胆红素(DBIL)均随胆道梗阻时间延长而升高,但DBL组AST、ALT在各时间点均较DBL+NAC组明显升高(P＜0.05),而TBIL、DBIL在这两组间无明显差异(P＞0.05)。DBL组和DBL+ NAC组TNF-α、NO浓度变化也随梗阻时间延长而升高,但DBL组较DBL+ NAC组TNF-α、NO浓度升高更明显(P＜0.05)。结论N-乙酰半胱氨酸能有效改善胆道梗阻所致肝损害,并有可能是通过下调肝组织中TNF-α、NO的表达这一途径实现的。     

Effective inhibition of nitric oxide production by aminoguanidine does not reverse hypotension in endotoxaemic rats

BACKGROUND: Excess production of nitric oxide (NO) by the inducible NO synthase (iNOS) has been implicated in the pathophysiology of septic shock. Using methaemoglobin (metHb) and the stable NO metabolite nitrate as markers of NO formation, we assessed the effect of iNOS blockade by aminoguanidine (AG) on hypotension and NO formation in endotoxaemic rats. METHODS: In 32 male Wistar rats under chloralose anaesthesia, MetHb (at 15 and 330 min, respectively) and plasma nitrate (at 330 min) were determined. Mean arterial pressure, heart rate and haematocrit were monitored. The LPS group (n=8) received bacterial endotoxin (LPS), 3 mg kg(-1) i.v. and was subsequently monitored for 5 h. At 2 h after LPS, the LPS+AG20 group (n=8) received AG, 5 mg kg(-1), and 5 mg kg(-1) h(-1) for the remaining 3 h. The LPS+AG100 group (n=8) instead received 25 mg kg(-1), followed by 25 mg kg(-1) h(-1). The NaCl group (n=8) was given corresponding volumes of isotonic saline. RESULTS: AG decreased the LPS-induced rise in plasma nitrate by about 50% in the LPS+AG20 group. MetHb levels, however, were not appreciably reduced by this dose. Both NO metabolites reached control levels after the higher dose of AG. LPS caused a progressive decrease in haematocrit. AG did not influence the LPS-induced hypotension, tachycardia or haemodilution. CONCLUSION: AG inhibited NO formation in a dose-dependent way. Yet, AG had no haemodynamic effects, suggesting a minor cardiovascular influence of iNOS in this endotoxin model, in parallel to what has been found in microbial sepsis.