NO与肝癌形成

目的研究一氧化氮(NO)在肝癌发生发展的作用。方法用免疫组化的方法对21例肝癌及癌旁组织中的3种一氧化氮合酶(NOS)及血管内皮细胞生长因子(VEGF)的表达进行原位检测和观察,结果紧邻癌细胞的肝硬化组织或慢性肝炎组织iNOS呈强阳性,远离癌组织的肝硬化组织或慢性肝炎组织多呈阴性或弥漫弱阳性;iNOS在周边癌组织及侵入纤维组织中的癌细胞呈阳性,癌组织核心多呈阴性或弥漫弱阳性。VEGF、nNOS的分布与iNOS相似。eNOS主要分布在肝癌细胞小血管壁内皮及其肌层组织。结论 NOS表达与肝组织癌变及肝癌侵润能力有关,与癌组织获得血管形成和转移表型有关。     

p53与一氧化氮的关系及其在肝癌发生中的作用

目的：研究p53与一氧化氮（NO）的关系及其在肝癌发生中的作用。方法：用免疫组化法对21全肝癌标本中p53、诱导型一氧化氮合酶（iNOS)及增殖细胞核抗原（PCNA）的表达进行原位检测和观察。结果：癌旁组织（距癌组织边缘＜1．5cm)iNOS表达多呈阳性,非癌组织（距离组织边缘＞1．5cm)多呈阴性或弥漫弱阳性;iNOS在周边癌组织及侵入纤维组织的癌细胞中呈阳性,癌组织核心多呈阴性或弥漫弱阳性。p53阳性率为47．6％。p53表达阳性区,iNOS表达也呈阳性。PCNA的表达与p53及iNOS一致。结论：肝癌旁组织中p53的表达与NO相关,与肝癌的发生、发展有关。     

NOS表达与肝癌增殖凋亡的关系研究

目的 研究一氧化氮合酶（NOS）表达与肝癌增殖凋亡的关系。方法 用免疫组化的方法对21例肝癌及癌旁组织中的诱导型一氧化氮合酶(iNOS)及增殖细胞核抗原（PCNA）的表达进行原位检测和观察,并与脱氧核糖核酸末端转移酶介导的制品末端标记（TUNEL）的凋亡细胞进行比较。结果 癌旁组织iNOS阳性,非癌组织多阴性或弥漫弱阳性;iNOS在周边的癌组织及侵入纤维组织中的癌细胞阳性,癌组织核心多阴结性或弥漫弱阳性。PCNA表达与iNOS相似,iNOS表达阳性区PCNA表达阳性率也高。TUNEL染色凋亡细胞阳率高区也是PCNA阳性率高区。结论 iNOS表达与肝癌的增殖凋亡有关,在肝癌的发生发展中起重要作用。     

一氧化氮在大鼠肝诱癌过程中的表达及其意义

目的：研究一氧化氮在大鼠肝诱癌过程中的表达及其意义。方法：用二乙基亚硝胺诱发大鼠肝癌。铜镉还原法测定诱癌不同阶段大鼠血中NO含量，用免疫组化方法对大鼠肝诱癌过程中肝硬化组织、癌及癌旁组织中诱导型一氧化氮合酶（iNOS)的表达进行原位检测和观察。结果：肝硬化组和肝癌组的血清中NO含量增加。正常对照组肝组织中未见iNOS阳性染色；诱癌硬化期肝组织iNOS染色阳性；肝癌形成期部分癌组织iNOS染色阳性，肝癌旁组织iNOS染色阳性。结论：二乙基亚硝胺诱发大鼠肝癌过程中一氧化氮起重要作用。     

一氧化氮合酶表达与肝癌增殖凋亡的关系研究

目的 研究一氧化氮合酶（ＮＯＳ）表达与肝癌增殖凋亡的关系。方法 用免疫组化的方法对２１世纪肝癌及癌旁组织中的诱导型一氧化氮合酶（ｉＮＯＳ）及增殖细胞核抗原（ＰＣＮＡ）的表达进行原位检测和观察,并与脱氧核糖核酸末端转移酶介导的缺口末端标记（ＴＵＮＥＬ）的凋亡细胞进行比较。结果 癌旁组织ｉＮＯＳ阳性,非癌组织多阴性或弥浊弱阳性;ｉＮＯＳ在周边的癌组织及侵入纤维组织中的癌细胞阳性,癌组织核心多阴性或弥温     

肝癌组织中诱导型一氧化氮合酶的表达

为了解诱导型一氧化氮合酶(induce nitricoxide synthase,iNOS)在肝癌发病中的作用,用免疫组织化学方法检测了肝癌组织中iNOS的表达。32例肝癌和10例正常肝组织进行HE染色、iNOS检测。10例正常肝组织内均未见iNOS阳性信号。肝癌组织iNOS表达高于正常对照肝组织(26/32vs 0/10,x2=31.328,P<0.01)。肝癌组织和癌旁肝组织均表达iNOS(23/32vs 25/32,x2=0.333,P=0.564)。iNOS通过多种途径参与肝癌的发病。     

缺氧性肺动脉高压大鼠肺iNOS mRNA及蛋白表达的实验研究

目的 :通过观察慢性缺氧大鼠肺组织诱生型一氧化氮合酶 (iNOS)表达的变化 ,探讨iNOS在肺动脉高压发病中的作用。方法 :运用血清学检验及免疫组织化学斑点杂交等方法观察血一氧化氮(NO)、肺组织iNOSmRNA及蛋白表达的变化。结果 :正常组大鼠NOS表达呈弱阳性 ,慢性缺氧后肺组织iNOSmRNA升高、蛋白NOS表达呈强阳性 ,血一氧化氮水平升高 (P <0 0 1)。结论 :诱生型一氧化氮合酶、一氧化氮的增加与慢性缺氧性肺动脉高压发病有关。     

诱生型一氧化合酶在肝癌中表达的意义

为证实诱生型一氧化氮合酶（iNOS）在肝癌发生、发展中的作用,采用免疫组化方法对50例原发性肝细胞癌（HCC）及癌旁组织（PCHT）中的iNOS表达情况进行了检测。结果显示,iNOS位于肝细胞细胞浆／膜中,在HCC中的表达强度变化较大,在PCHT中呈强表达;正常肝组织中呈弱表达;淋巴细胞中无表达。证实iNOS的异常表达可导致HCC患者体内NO生成异常及机体对癌细胞的免疫异常。提示提高体内NO合成,可望提高HCC患者的抗癌能力。     

人胃癌组织中一氧化氮合酶的表达

目的探讨NOS与胃癌的关系.方法用NADPH-d组织化学法测定了正常胃组织、癌旁组织和癌组织中一氧化氮合酶(NOS)表达水平.结果正常胃组织中粘膜上皮细胞、各种有分泌功能的细胞及肌层神经纤维中均有NOS表达,测一个视野NOS阳性细胞的平均灰度,正常胃组织为112、癌旁组织为120、胃癌组织为145.各组间差异有显著意义.表明正常胃组织NOS活性最高,胃癌组织NOS活性最低.结论①正常胃组织有广泛的NOS分布,提示NO对维持正常胃功能具有重要作用;②胃粘膜细胞癌变过程中,NOS活性明显降低,提示NOS活性与胃粘膜细胞癌变有高度相关性.     

胎盘型谷胱甘肽S-转移酶在人乙型肝炎肝硬化肝癌中的表达及其意义

应用抗胎盘型谷胱甘肽S-转移酶(GST-π)单克隆抗体,PAP免疫组化染色,对肝癌、肝硬化及乙型肝炎进行了研究。42例肝癌(包括肝细胞型肝癌和胆管型肝癌),GST-π阳性率为85.7%,乙型肝炎和肝硬化阳性率分别为30%,65%。高分化肝细胞癌和胆管型肝癌全部呈阳性表达,而低分化肝细胞癌呈阴性表达。正常肝组织GST-π仅在部分胆管上皮呈弱阳性表达。结果提示:GST-π可能是高分化肝细胞型肝癌和胆管型肝癌的标志酶,GST-π用来鉴别低分化肝细胞癌和胆管型肝癌可能有一定价值。     

The plasma level of nitric oxide and the expression of inducible nitric oxidesynthase in human hepatocellular carcinoma

AIM To study the relationship between nitric oxide (NO), nitric oxide synthase (NOS) and humanhepatocellular carcinoma (HCC).METHODS Plsama NO2-/NO3- was measured by Griess reaction in 122 patients with chronic hepatitis(CH) and compensated liver cirrhosis (LC), among which 62 patients were complicated with HCC(CH = 28, LC = 34), and the rest 60 patients were not (CH = 29, LC = 31). Thirty healthy persons served asnormal controls (NC). There were no prominent differences among the groups in sex, age and the ratio ofCH to LC. The expression of inducible nitric oxide synthase (iNOS) in HCC (n = 40), CH (n = 30) and LC(n = 30) samples obtained from liver biopsy or operation was compared with that in normal liver tissues byusing immunohistochemistry. Ten normal liver tissue samples obtained from liver operation served as normalcontrols. The samples were fixed in formalin and embeded in paraffin. Anti-iNOS antibody (Santacruzcompany) was served as antibody-Ⅰ in immunohistochemical assay of iNOS in tissue.RESULTS Plasma NO2-/NO3- level in normal was 11.5 μmol/L±4.2μmol/L. The plasma level ofNO2 /NO3- in CH (58.6±17.4 μmol/L) and LC (38.7±10.6μmol/L) accompanied with HCC wasnotably higher than in those patients without HCC (CH: 24.8±9.4 μmol/L; LC: 22.3±8.7μmol/L,t=2.901, 2.756, P<0.01). Plasma NO2-/NO3- level in HCC accompanied with CH was significantlyhigher than in those accompanied with LC ( t = 2.216, P<0.05). Positive rate of iNOS in HCC, CH and LCwas 95%, 93% and 57% respectively. iNOS was not expressed in normal liver tissues. The expression level ofiNOS in HCC (χ2=17.4, P<0.001) and CH (χ2=11.64, P<0.025) was much higher than in LC.CONCLUSION Plasma NO2 / NO3- level significantly increased in patients with HCC and theimmunohistochemical staining of iNOS was positive. This suggests that the liver secrets NO in the higherlevel may participate in the carcinogenesis and progression of HCC.     

iNOS/NO体系在慢性肝炎及肝硬化患者中的表达

目的:探讨内源性NO在慢性病毒性肝炎-肝硬化发展进程中的作用.方法:采用硝酸还原酶法比色测定外周静脉及门静脉血浆中iNOS活性,并用免疫组织化学和RT-PCR方法观察肝组织iNOS蛋白及RNA的表达.结果:在慢性肝炎患者及肝硬化患者门静脉血与外周血中iNOS活性与对照组相比均明显升高(F=102.793,25.052,P<0.01),且门静脉血iNOS活性增高更为明显.慢性肝炎组及肝硬化组中iNOS蛋白的灰阶值均低于正常对照组(F=46.796,P<0.05),表明iNOS表达增强.慢性肝炎组、肝硬化组iNOS mRNA的表达均分别显著高于正常对照组(F=26.832,P<0.01),且随着肝脏病变的加重表达逐渐增加.结论:iNOS/NO体系在慢性肝炎-肝硬化发生发展中起着保持血管舒张状态的重要作用.     

Diabetic ketosis activates lymphomonocyte-inducible nitric oxide synthase.

AIMS: Inappropriate production of nitric oxide (NO) may be responsible for the haemodynamic disturbances of diabetic ketoacidosis. We investigated whether this metabolic condition is associated with increased plasma nitrate (the stable oxidation product of NO) levels and NO synthase gene expression in lymphomonocytes. RESEARCH DESIGN AND METHODS: Plasma nitrate concentrations, lymphomonocyte-inducible nitric oxide synthase (iNOS) gene expression, tumour necrosis factor-alpha (TNF-alpha) and soluble thrombomodulin were measured in 11 Type 1 diabetic patients at baseline, during mild ketosis and after euglycaemia was re-established. RESULTS: During diabetic ketosis plasma nitrate concentrations were higher (18 (16-21) vs. 9 (7-11) micro mol/l; (95% lower-upper confidence interval) P < 0.05) than at baseline. At baseline lymphomonocyte iNOS mRNA expression and iNOS protein levels were undetectable, but in ketosis both were increased (both at P < 0.0001). After recovery from ketosis, NO3 concentration, iNOS mRNA, and iNOS expression (270 +/- 36%, mean +/- sd) decreased but not significantly. No significant changes were observed in either TNF-alpha or soluble thrombomodulin levels between the three conditions. CONCLUSIONS: Diabetic ketosis is associated with increased nitrate levels and the activation of iNOS expression in circulating lymphomonocytes, but it does not affect either the proinflammatory cytokine TNF-alpha or a marker of endothelial dysfunction such as thrombomodulin. Our data support the hypothesis that, during diabetic ketosis, alterations in NO homeostasis are present in circulating lymphomonocytes.     

Production of reactive oxygen species and expression of inducible nitric oxide synthase in rat isolated Kupffer cells stimulated by Leptospira interrogans and Borrelia burgdorferi

AIM: To evaluate the production of reactive oxygen species (ROS) and the expression of inducible nitric oxide synthase (iNOS) in rat isolated Kupffer cells (KCs) stimulated by Leptospira interrogans and Borrelia burgdorferi. METHODS: Rat Kupffer cells were separated by perfusion of the liver with 0.05% collagenase, and purified by Percoll gradients. Purified Kupffer cells were tested in vitro with alive L. interogans and B. burgdorferi preparations. The production of ROS was determined by chemiluminescence, whereas iNOS protein expression was evaluated by Western blot assay using anti-iNOS antibodies. RESULTS: B. burgdorferi and to a less extent L. interrogans induced ROS production with a peak 35 min after infection. The chemiluminescence signal progressively diminished and was undetectable by 180 min of incubation. Leptospirae and borreliae induced an increased iNOS expression in Kupffer cells that peaked at 6 hours and was still evident 22 h after infection. CONCLUSION: Both genera of spirochetes induced ROS and iNOS production in rat Kupffer cells. Since the cause of liver damage both in leptospiral as well as in borrelial infections are still unknown, we suggest that leptospira and borrelia damage of the liver can be initially mediated by oxygen radicals, and is then maintained at least in part by nitric oxide.