原发性肝癌一氧化氮及一氧化氮合酶的研究

探讨一氧化氮 (Nitricoxide,NO)及诱导型一氧化氮合酶 (induciblenitricoxidesynthase ,iNOS)与原发性肝癌 (HCC)间的关系 ,用Griess反应测定 16 2例患者的血浆亚硝酸盐 /硝酸盐 (NO-2 /NO-3 )水平 ,其中HCC82例 ,非HCC80例 ,健康对照 36名。用免疫组化法检查组织中iNOS的含量 ,取正常肝脏组织 2 0例作对照 ,慢性肝炎 (CH)和肝硬化 (LC)的肝脏组织各 40例 ,HCC组织 48例。结果显示 ,正常人血浆NO-2 /NO-3 含量为 16 .8±4.9μmol/L,有HCC的CH(6 3 .4± 18.2 μmol)和LC(42 .2± 11.5 μmol/L)明显高于非HCC的患者 (CH :2 8.5±8.7μmol/L;LC :2 4.7± 6 .2 μmol/L .P <0 .0 1) ,患CH的HCC患者血浆NO-2 /NO-3 水平明显高于LC基础上的HCC患者 (P <0 .0 5 )。正常肝组织iNOS阴性 ,LC有 2 5例 (6 2 .5 % )阳性 ,CH 36例 (90 % )阳性 ,HCC 46例 (95 8% )阳性 ;且CH (P <0 .0 2 5 )及HCC(P <0 .0 0 1)的表达水平明显高于LC。提示HCC患者有NO分泌的增加 ,NO可能参与HCC的发生发展的病理过程     

iNOS在乙肝肝硬化组织中的表达及与肝细胞增殖凋亡的关系

采用免疫组化法检测诱导型一氧化氮合酶（iNOS）及增殖细胞核抗原（PCNA））在乙肝肝硬化组织（观察组）和正常肝组织（对照组）中的表达,并与TUNEL染色凋亡细胞进行相关性分析。结果观察组iNOS表达阳性率100%,对照组未见iNOS表达,两组iNOS水平有显著差异（P〈0.05）;PCNA阳性细胞数和凋亡细胞数均与iNOS表达呈正相关,P〈0.01。认为iNOS可能参与了肝硬化的发生、发展,机理为促进细胞增殖。     

HIF-1α与iNOS在胃癌组织中的表达及意义

目的研究胃癌组织中缺氧诱导因子1α（HIF-1α）和诱导型一氧化氮合酶（iNOS）的表达及意义,探讨HIF-1α与iNOS的相关性。方法胃癌手术标本95例,采用免疫组化法检测HIF-1α和iNOS在胃癌组织和癌旁正常胃黏膜组织中的表达。结果HIF-1α和iNOS在胃癌组织中的表达水平明显高于癌旁正常组织（P均〈0．05）;HIF—1α和iNOS的表达与胃癌的TNM分期、浸润程度和淋巴结转移有关（P〈0．05）;HIF-1α与iNOS正相关（r=0．710,P〈0．05）。结论HIF-1α与iNOS联合检测可作为反映胃癌生物学行为的指标。     

肝硬化时各系统内源性NO变化的研究进展

作为终末期肝病的肝硬化常常引起多系统损害。内源性NO是一种具有多种病理生理学效应的活性介质,它对疾病发生发展的影响是近年来的研究热点,本文就肝硬化时各系统内源性NO变化的研究进展作一综述。     

iNOS在老年NSCLC患者中表达及其与患者临床病理特征和预后的关系

目的 探讨诱导型一氧化氮合酶(iNOS)在老年非小细胞肺癌(NSCLC)患者中表达及其与患者临床病理特征和预后的关系。方法 选择行手术切除的老年NSCLC患者100例,取患者癌组织及其癌旁正常组织。收集患者临床病理特征,包括性别、吸烟史、肿瘤大小、病理类型、组织学分化、TNM分期和淋巴结转移。比较癌组织与癌旁正常组织iNOS阳性表达情况;比较不同病理特征iNOS阳性表达情况;生存曲线分析NSCLC患者iNOS表达与预后关系;采用多因素Logistic回归分析影响预后独立危险因素。结果 癌组织iNOS阳性表达显著高于癌旁组织(P<0.001)。不同性别、吸烟史、肿瘤大小、组织学分化iNOS阳性表达比较差异无统计学意义(P>0.05);Ⅲ～Ⅳ期iNOS阳性表达显著高于Ⅰ～Ⅱ期(P<0.05);淋巴结转移iNOS阳性表达显著高于无淋巴结转移(P<0.05)。NSCLC患者iNOS阳性表达总生存期(OS)时间显著短于iNOS阴性表达(P<0.05)。经多因素Logistic回归分析显示,Ⅲ～Ⅳ期、淋巴结转移和iNOS阳性表达为影响预后独立危险因素。结论 老年NSC...     

PAF受体拮抗剂对内毒素血症幼年大鼠胃黏膜NO含量及iNOS表达的影响

目的:探讨一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)及PAF受体拮抗剂在内毒素(LPS)腹腔注射诱导的幼年大鼠急性胃黏膜损伤中的作用.方法:Wistar大鼠, 随机分为对照组、LPS组、PAF受体拮抗剂预防组和治疗组. 用内毒素(O55:B5脂多糖)5 mg/kg ip制备幼年大鼠内毒素血症模型. 预防组和治疗组分别于内毒素ip前及ip后0.5 h, 应用血小板活化因子(PAF)受体拮抗剂BN52021(GinkgolideB)5 mg/kg ip, 同等量生理盐水ip为对照组. 于LPS注射后1.5,3, 6, 24, 48, 72 h处死动物, 大体及光学显微镜下观察胃黏膜损伤情况, 采用硝酸还原酶的化学比色法测定胃黏膜NO含量;免疫组织化学S-P方法测定胃黏膜iNOS蛋白的表达, 半定量RT-PCR法测定胃黏膜iNOS mRNA的表达.结果:LPS组6 h胃黏膜损伤最重, 黏膜内有出血, 核碎裂、固缩, 凋亡小体出现;预防组和治疗组改变轻微. LPS组腹腔注射内毒素后6 h胃黏膜NO含量最高, 此时LPS组较对照组NO含量明显增高(84.37±5.44 vs 37.37±1.90,P<0.01), 预防组和治疗组(40.07±3.42, 48.63±3.24)较LPS组明显降低( P<0.01);预防组与治疗组较对照组明显增高( P<0.05). 对照组胃黏膜组织未见iNOS蛋白及mRNA的表达;LPS组腹腔注射内毒素后1.5 h胃黏膜组织胞质iNOS蛋白表达, 6 h明显增高, 24 h最高, 48 h下降, 72 h仍未恢复正常;预防组和治疗组3 hiNOS蛋白表达, 6 h明显增高, 48 h下降, 72 h同对照组. iNOS mRNA水平的表达变化与iNOS蛋白表达变化趋势相同.结论:PAF受体拮抗剂可下调iNOS mRNA表达水平, 减少iNOS蛋白表达, 使NO含量下降.从而使NO和iNOS对胃黏膜发挥保护作用.     

MIF和iNOS在胎膜早破产妇胎膜组织中的表达及意义

目的探讨巨噬细胞移动抑制因子（MIF）与诱导型一氧化氮合酶（iNOS）在胎膜早破（PROM）发生、发展中的作用。方法研究对象为90例剖宫产的产妇;其中早产胎膜早破（p PROM）30例（p PROM组）,足月胎膜早破（t PROM）30例（t PROM组）,正常足月妊娠30例（正常组）。三组均于剖宫产术中胎盘娩出后于距离胎膜破口〉2 cm处取1 cm×1 cm的胎膜组织制作切片。采用HE染色方法检测绒毛膜羊膜炎（中性粒细胞中、重度浸润）发生情况;采用免疫组化SP法检测MIF、iNOS表达;分析三组及绒毛膜羊膜炎、非绒毛膜羊膜炎产妇MIF、iNOS表达情况。结果三组共检出绒毛膜羊膜炎27例;MIF和iNOS主要表达于羊膜上皮细胞的胞质。p PROM组和t PROM组MIF表达强度明显高于对照组（P〈0.01）;p PROM组iNOS表达强度明显高于对照组（P〈0.01）;绒毛膜羊膜炎产妇胎膜中MIF和iNOS表达强度明显高于非绒毛膜羊膜炎产妇（P〈0.05）。结论 PROM产妇胎膜组织中MIF、iNOS表达上调。MIF、iNOS高表达可促进胎膜早破及绒毛膜羊膜炎的发生。     

吡喹酮对血吸虫病肝纤维化小鼠血清NO及肝内iNOS活性的影响

目的:研究吡喹酮治疗血吸虫病肝纤维化小鼠前后血清一氧化氮及肝内诱导型一氧化氮合酶活性的变化。方法:制备血吸虫病肝纤维化小鼠模型,应用免疫组化染色方法和多媒体彩色病理图文分析系统定量观察吡喹酮治疗前后肝脏诱导型一氧化氮合酶含量的变化,并通过化学还原反应检测血清一氧化氮的水平变化。结果:吡喹酮治疗组肝内iNOS和血清NO的含量均低于感染组,P均小于0.01。结论:吡喹酮可显著降低血吸虫病肝纤维化小鼠肝脏诱导型一氧化氮合酶活性及血清NO含量而发挥抗血吸虫病肝纤维化的作用。     

慢性肝炎和肝硬化患者血浆一氧化氨与GMP—140水平及其意义

目前 ,不少学者认为一氧化氮 (ＮＯ)在肝脏疾病中起重要作用 ,ＮＯ水平与各种肝脏疾病程度有关。近年来许多研究还表明 ,肝脏疾病的发生、发展亦与粘附分子的表达有关 ,抗细胞粘附治疗正发展成为治疗肝细胞坏死的新战略。我们测定了慢性肝炎及活动性肝硬化患者血浆ＮＯ-2 /ＮＯ-3 和可溶性α 颗粒膜蛋白 (ＧＭＰ 14 0 )水平 ,以探讨其临床意义。材料与方法一、研究对象全部病例为本院附属医院 1997年～ 1998年收治的住院患者 ,诊断按 1995年北京全国传染病与寄生虫病学术会议修订的标准。 12 6例患者 ,男 83例 ,女 43例 ,年龄 19～ 75岁 ,平…     

Temporal expression of hepatic inducible nitric oxide synthase in liver cirrhosis

AIM: Nitric oxide (NO) has been implicated in the pathogenesis of liver cirrhosis. We have found inducible nitric oxide synthase (iNOS) can be induced in hepatocytes of cirrhotic liver. This study further investigated the temporal expression and activity of hepatic iNOS in cirrhosis development. METHODS: Cirrhosis was induced in rats by chronic bile duet ligatjon (BDL). At different time points after the operation, samples were collected to examine NO concentration, liver function, and morphological changes. Hepatocytes were isolated for determination of iNOS mRNA, protein and enzymatic activity. RESULTS: Histological examination showed early cirrhosis 1-2 wk after BDL, with advanced cirrhosis at 3-4 wk. Bilirubin increased dramatically 3 d after BDL, but decreased by 47% on d 14. Three weeks after BDL, it elevated again. Systemic NO concentration did not increase significantly until 4 wk after BDL, when ascites developed. Hepatocyte iNOS mRNA expression was identified 3 d after BDL, and enhanced with time to 3 wk, but reduced thereafter. iNOS protein showed a similar pattern to mRNA expression. iNOS activity decreased from d 3 to d 7, but increased again thereafter till d 21. CONCLUSION: Hepatic iNOS can be induced in the early stage, which increases with time as cirrhosis develops. lts enzymatic activity is significantly correlated with protein expression and histological alterations of the liver, but not with systemic NO levels, nor with absolute values of liver function markers.     

诱导型一氧化氮合酶在反流性食管炎患者食管黏膜中的表达

目的探讨iNOS在RE发病机制中的作用。方法本研究收集77例反流性食管炎（RE）及20例对照组的食管下段括约肌处的食管黏膜。以免疫组化SP法结合图像分析检测食管黏膜中诱导型一氧化氮合酶（iNOS）的免疫反应阳性产物的平均光密度值。结果免疫组化结果显示：iNOS在RE患者中的表达明显强于对照组（P〈0.05）。结论iNOS在RE患者中的食管黏膜中的表达异常,表明NO在RE的发病机制中可能起重要作用。     

诱导型和内皮型一氧化氮合酶基因多态性与肝硬化门静脉高压的相关性

目的探讨诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)基因多态性与肝硬化门静脉高压的相关性。方法采用病例对照和聚合酶链反应-限制性片段长度多态性技术,检测106 例乙型肝炎后肝硬化患者(其中门静脉高压症65例)和108名健康对照者iNOS基因启动子-969G→C多态性及eNOS基因第七外显子894G→多态性,比较等位基因及基因型频率,并进行综合分析。结果在iNOS启动子969G→C多态性中,门静脉高压症组C等位基因和GC基因型频率比对照组显著升高(x2- 5.93,P<0.05)。GC基因型启动子的活性比GG基因型活性强。在eNOS的894G→T多态性中,T等位基因频率明显高于对照组(x2-3.91,P<0.05)。采用Logistic多元回归显示iNOS基因启动子-969G→C多态性及cNOS的894G→T多态性是门静脉高压症新的危险因素。结论iNOS启动子969G→C多态性和eNOS的894G→T多态性与门静脉高压症相关,是形成门静脉高压症新的危险因素。iNOS启动子-969G→C多态性可导致该基因启动子功能活性增强。     

Glycyrrhizinate reduces portal hypertension in isolated perfused rat livers with chronic hepatitis

AIM:To investigate the effects of diammonium glycyrrhizinate(Gly)on portal hypertension(PHT)in isolated portal perfused rat liver(IPPRL)with carbon tetrachloride(CCl4)-induced chronic hepatitis.METHODS:PHT model was replicated with CCl4 in rats for 84 d.Model was identified by measuring the ascetic amounts,hepatic function,portal pressure in vivo,splenic index,and pathological alterations.Inducible nitric oxide synthase(iNOS)in liver was assessed by immunohistochemistry.IPPRLs were performed at d0,d28,d56,and d84.After phenylephrine-induced constriction,Gly was geometrically used to reduce PHT.Gly action was expressed as median effective concentration(EC50)and area under the curve(AUC).Underlying mechanism was exploited by linear correlation between AUC values of Gly and existed iNOS in portal triads.RESULTS:PHT model was confirmed with ascites,splenomegaly,serum biomarkers of hepatic injury,and elevated portal pressure.Pathological findings had shown normal hepatic structure at d0,degenerations at d28,fibrosis at d56,cirrhosis at d84in PHT rats.Pseudo lobule ratios decreased and collagen ratios increased progressively along with PHT development.Gly does dose-dependently reduce PHT in IPPRLs with CCl4-induced chronic hepatitis.Gly potencies were increased gradually along with PHT development,characterized with its EC50at 2.80×10-10,3.03×10-11,3.77×10-11and 4.65×10-11mol/L at d0,d28,d56and d84,respectively.Existed iNOS was located at hepatocyte at d0,stellate cells at d28,stellate cells and macrophages at d56,and macrophages in portal triads at d84.Macrophages infiltrated more into portal triads and expressed more iNOS along with PHT development.AUC values of Gly were positively correlated with existed iNOS levels in portal triads.CONCLUSION:Gly reduces indirectly PHT in IPPRL with CCl4-induced chronic hepatitis.The underlying mechanisms may relate to rescue NO bioavailability from macrophage-derived peroxynitrite in portal triads.     

iNOS在肺癌中的表达及其与肿瘤病理,临床分期的关系

采用逆转录－聚合酶链反应技术检测４０例不同肿瘤病理，临床分期的肺癌组织中诱生型一氧化氮合酶ｍＲＮＡ的表达情况。结果表明，肺癌组ｉＮＯＳｍＲＮＡ阳性表达率为７２．５０％，较对照组明显增高。     

氟骨症患者血清一氧化氮含量和外周血白细胞诱导型一氧化氮合酶mRNA表达的意义

目的　探讨氟骨症患者患者外周血白细胞诱导型一氧化氮合酶(induciblenitricoxidesynthaseiNOS)mRNA表达的意义。方法　选择3 5例氟骨症患者患者及3 0例正常人的外周血白细胞。iNOS -mRNA表达采用原位杂交方法。血浆一氧化氮(nitricoxideNO)水平采用硝酸还原酶比色法测定。结果　正常人外周血白细胞未见iNOS -mRNA表达,而氟骨症患者外周血白细胞iNOS -mRNA表达高度增强,其阳性率达40 .82 %。AR组血清NO水平显著高于对照组(t =2 7.89,P <0 .0 1)。结论　氟骨症患者病人血清NO水平与外周血白细胞iNOS -mRNA表达高度增强有关。提示iNOS-NO通路在氟骨症的发病过程中可能起重要作用。本研究为检测体内某些信号提供了简便易行的原位杂交试验方法。     

抑郁症对大鼠结肠iNOS、NO和COX-2的影响

目的　观察抑郁症对大鼠结肠组织诱导型一氧化氮合酶 (iNOS)、一氧化氮 (NO)含量和环氧合酶 2 (COX 2 )表达的影响。方法　制备经典的大鼠抑郁模型。采用生物化学试剂盒检测大鼠结肠组织中iNOS和NO的含量 ,免疫组化SP方法检测结肠组织COX 2的表达。结果　抑郁模型大鼠结肠组织中iNOS和NO的含量均升高 (P <0 .0 1) ;COX 2在抑郁大鼠结肠中表达升高 (P<0 .0 1)。结论　抑郁症对大鼠结肠组织的损伤可能通过改变iNOS和NO的含量 ,及COX 2的表达实现的     

New progress in understanding roles of nitric oxide during hepatic ischemia-reperfusion injury

Hepatic ischemia-reperfusion injury (HIRI) is a major clinical cause of morbidity and mortality in liver surgery and transplantation. Many studies have found that nitric oxide (NO) plays an important role in the HIRI and its increase or decrease can affect the progression and outcome of HIRI. However, the role of NO in HIRI is controversial and complicated. NO derived by endothelial NO synthase (eNOS) shows a protective role in HIRI, while excessive NO derived by inducible NO synthase (iNOS) accelerates inflammation and increases oxidative stress, further aggravating HIRI. Nevertheless, the overexpression of eNOS may exacerbate HIRI and iNOS-derived NO in some cases reduces HIRI. Here we review the new progress in the understanding of the roles of NO during HIRI: (1) NO possesses different roles in HIRI by increasing NO bioavailability, down-regulating leukotriene C4 synthase, inhibiting the activation of the nuclear factorκB (NFκB) pathway, enhancing cell autophagy, and reducing inflammatory cytokines and reactive oxygen species (ROS). And NO has both protective and deleterious effects by regulating apoptotic factors; (2) eNOS promotes NO production and suppresses its own overexpression, exerting a hepatoprotective effect reversely. Its activation is regulated by the PI3K/Akt and KLF2/AMPK pathways; and (3) iNOS derived NO mainly has deteriorating effects on HIRI, while it may have a protective function under some conditions. Their expression should reach a balance to reduce the adverse side and make NO protective in the treatment of HIRI. Thus, it can be inferred that NO modulating drugs may be a new direction in the treatment of HIRI or may be used as an adjunct to mitigate HIRI for the purpose of protecting the liver.     

iNOS Expression in Oral and Gastrointestinal Tract Mucosa

It is known that the overproduction of nitric oxide (NO) by nitric oxide synthase (NOS) occurs during the progression of various inflammatory diseases in intestinal tract. NOS inhibitors or inducible nitric oxide synthase (iNOS) gene expression inhibitors should be considered as potential anti-inflammatory agents, as NO synthesized by iNOS is related to various pathophysiological processes including inflammation. In order to understand the relationship between iNOS and pathological reactions such as the inflammatory process and malign transformation clearly, the existence and amount of constitutive expression should be determined. It is crucial to comprehend the harmful and protective amounts of iNOS expressions in order to clarify the relationship between iNOS and pathological processes. Evidently, only after this inspection is it possible to utilize iNOS as a marker and treatment instrument during the diagnosis and treatment of malign transformation and the inflammatory process.     

内皮型一氧化氮合酶基因G894T突变与肝硬化门脉高压的相关性研究

目的　探讨内皮型一氧化氮合酶 (endothelialnitricoxidesynthase ,eNOS)基因第 7外显子G894T点突变与肝硬化门脉高压症之间的关系。方法　采用病例对照和聚合酶链反应 限制性片段长度多态性 (PCR RFLP)方法 ,检测10 6例乙型肝炎后肝硬化患者和 10 8名健康对照者eNOS基因第 7外显子G894T点突变频率和外周血NO 2 /NO 3 含量 ,比较各组间基因型频率与等位基因频率。结果　①中国汉族正常人eNOS基因G894T突变GG、GT和TT基因型频率分别为 86.1%、11.1%和 2 .8%;G、T等位基因频率分别为 91.7%和 8.3 %。②乙型肝炎后肝硬化组GT TT基因型频率高于对照组 ( 2 4.5 %比 3 .9%) ,差异有显著性。③门脉高压症组T等位基因频率明显高于对照组 ,差异有显著性( 17.7%比 8.3 ,P <0 .0 5 ) ,相关分析呈正相关 (r =0 .2 )。携带T等位基因者发生门脉高压症的危险性高于非T等位基因携带者 1.76倍 (OR =2 .76)。结论　eNOS基因第 7外显子G894T突变与肝硬化门脉高压症形成相关 ,T等位基因可能是中国人群门脉高压症的遗传易感基因型