冠心康对实验性大鼠急性心肌缺血的保护作用

目的：探讨冠心康颗粒对大鼠急性心肌缺血的保护作用及其机理。方法：雄性大鼠32只，随机分为4组，每组8只，预防给药7d。灌胃结束后，采用在体大鼠冠状动脉左前降支结扎的方法建立心肌缺血模型，观察冠心康颗粒对大鼠急性心肌缺血后心肌中NO和NOS含量的影响，用免疫组化法检测Bcl-2蛋白在心肌中的表达。结果：模型组、冠心康组、地奥组与假手术组比较，心肌组织NO及NOS含量均有明显降低（P〈0．05）；而与模型组相比，冠心康组、地奥组的NO及NOS含量均有明显的恢复（P〈0．05），提示冠心康颗粒可明显提高大鼠急性心肌缺血后心肌中NO和NOS含量（P〈0．005，0．005），同时增加Bcl-2蛋白的表达，减少细胞凋亡。结论：冠心康颗粒对大鼠急性心肌缺血具有保护作用，可能与其提高心肌中NO和NOS含量，增加Bcl-2蛋白的表达有关。     

加味小柴胡汤对顺铂诱导大鼠肝BRL细胞过氧化损伤的保护作用

目的:探讨加味小柴胡汤对顺铂诱导大鼠肝BRL细胞氧化损伤的保护作用。方法:采用体外细胞培养方法,观察加味小柴胡汤对顺铂诱导的BRL细胞生长、SOD、GSH-px、NOS活性、MDA、GSH、T-AOC含量的影响。结果:顺铂各剂量处理细胞OD值皆显著下降,且随剂量而加重;加味小柴胡汤大小剂量细胞OD值皆明显升高;顺铂组SOD、GSH-px、T-AOC活性、GSH含量均明显降低;MDA含量及NOS活性明显升高;两实验组SOD、GSH-px、T-AOC及GSH均较顺铂组升高,MDA及NOS降低,而以1组效果更显著。结论:顺铂可明显抑制肝细胞生长,诱致细胞过氧化损伤,加味小柴胡汤可提高细胞生长率,其作用与其明显的抗氧化能力有关,且有一定的量效关系。     

枸杞多糖对染铅小鼠海马区NO和NOS活性的影响

目的:探讨枸杞多糖对染铅小鼠海马区NO和NOS活性的影响。方法:腹腔注射Pb(AC)2使小鼠染铅,枸杞多糖于染铅后第二天灌胃给药,于第6周后测全血铅浓度和小鼠海马区NO含量及NOS活性。结果:枸杞多糖组血铅浓度显著低于染铅模型对照组;枸杞多糖3.6g/kg组与染铅模型对照组比较,NO浓度和NOS活性显著升高。结论:枸杞多糖可促进小鼠体内铅的排出,提高小鼠海马区NO含量和NOS活性。     

药物血清对缺氧/复氧心肌细胞Ca~(2+)及NOS-NO系统的相关性研究

目的探讨双参通冠方药物血清对培养心肌细胞缺氧/复氧损伤的Ca2+超载、NOS-NO系统的影响。方法培养心肌细胞,建立缺氧/复氧损伤模型。运用血清药理学方法研究双参通冠方药物血清对缺氧/复氧心肌Ca2+超载、NOS-NO系统的影响。荧光分光光度法测定心肌细胞胞质[Ca2+]i,比色法检测心肌细胞培养上清NOS活性、NO含量。结果缺氧/复氧后[Ca2+]i增高,总NOS(T-NOS)及诱导型NOS(iNOS)活性增高,双参通冠方药物血清能降低缺氧/复氧心肌细胞[Ca2+]i,并能降低T-NOS、iNOS活性和NO含量(P<0.05)。结论双参通冠方药物血清可抑制缺氧/复氧损伤时心肌细胞Ca2+超载及NOS-NO系统的激活。     

乙醇诱导人脐静脉内皮细胞损伤及ADMA/NOS/NO信号机制

目的探讨乙醇诱导人脐静脉内皮细胞损伤效应及ADMA/NOS/NO信号机制。方法用不同浓度的乙醇与人脐静脉内皮细胞共培养,观察细胞形态和活性,通过检测细胞上清液丙二醛（MDA）、非对称性二甲基精氨酸（AD-MA）、一氧化氮（NO）的含量和一氧化氮合酶（NOS）的活性等指标,观察乙醇对人脐静脉内皮细胞损伤的量-效关系（乙醇处理细胞24 h）和时-效关系（终浓度为100 mmol/L的乙醇分别于处理0、3、6、12、24、48 h后观察检测指标）。实验分为正常对照组（加入与处理组等体积的D-Hank’s液,乙醇浓度为0 mmol/L）、乙醇处理组（终浓度分别为25、50、100、200mmol/L的乙醇）、乙醇＆VitC组（乙醇终浓度为100 mmol/L,VitC终浓度为100 mg/L）和阳性对照组（终浓度为500μmol/L的过氧化氢）。结果50-200 mmol/L乙醇能显著改变细胞形态、降低细胞活性（OD值：0.91±0.10-0.58±0.04,细胞生长抑制率：0.00±0.00-35.57±3.13,P〈0.01）;能显著性诱导内皮细胞MDA升高（258.72±7.36-524.07±8.25,P〈0.01）。50-200 mmol/L的乙醇还能显著性诱导：内皮型一氧化氮合酶（eNOS）活性下降（6.27±0.10-0.47±0.23,P〈0.05）及总NOS活性下降（10.69±0.54-3.77±0.32,P〈0.05）;NO含量降低（191.61±7.96-88.38±5.07,P〈0.05）。而且上述生物学效应均具有显著的时间依赖性,各处理组间两两比较也具有显著性（P〈0.05或P〈0.01）。100 mmol/L乙醇处理时细胞诱导型一氧化氮合酶（iNOS）活性及ADMA含量最高,且分别在处理后24 h到达最高峰。VitC均能扭转上述生物学效应,且具有显著性（P〈0.01）。结论50-200 mmol/L的乙醇能显著诱导人脐静脉内皮细胞损伤;ADMA在乙醇诱导人脐静脉内皮细胞损伤中有着重要作用。     

PPAR-α激动剂对大鼠内皮功能不全的作用及机制探讨

目的:观察PPAR-α激动剂对高脂血症模型大鼠内皮功能不全的作用,并初步探讨其作用机制。方法:称重后,32只SD雄性大鼠随机分成4组,每组8只:正常对照组(A组);正常+非诺贝特治疗组(B组);高脂组(C组);高脂+非诺贝特治疗组(D组)。每周称体重,按体重调整饲料及药物量。12周后,分别检测血脂4项(TC、TG、LDL-c、HDL-c)及血清中NO含量。分离胸主动脉,检测组织匀浆中总NOS活性及离体血管环的舒缩功能。结果:在调脂方面,PPAR-α激动剂非诺贝特能显著降低TG,并能在一定程度上降低LDL,同时显著升高HDL的作用。此外,PPAR-α激动剂非诺贝特具有显著增加总NOS活性及升高NO含量的作用,能显著改善内皮依赖性舒张功能,其作用并不完全依赖于血脂的降低。结论:PPAR-α激动剂非诺贝特能显著增加总NOS活性从而升高NO含量及改善血管内皮依赖性舒张功能,对内皮功能不全起到预防及治疗的作用,这可能是来自于调脂以外的作用。     

柚皮苷对经TNF-α诱导体外培养兔关节软骨细胞增殖及SOD NOS影响

目的:观察骨碎补柚皮苷对经肿瘤坏死因子(TNF-α)诱导体外培养兔关节软骨细胞增殖的影响.及对体外培养免关节软骨细胞超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)的影响.方法:原代分离软骨细胞经DMEM(含10?S)培养得到贴壁细胞并传代,行甲苯胺蓝染色及Ⅱ型胶原免疫组化鉴定;传代细胞经TNF-α诱导,用MTT法测增殖率,以及检测上清液中SOD,NOS的活性.结果:100ug/mL,50ug/mL,25ug/mL柚皮苷组对软骨细胞的增殖有明显的促进作用,与空白对照组相比有显著性差异(P<0.05),其中25ug/mL的促进作用最强,A值最高.各浓度组柚皮苷均可显著提高软骨细胞的SOD活性.与空白对照组和实验对照组相比均有显著性差异(P<0.05).不同浓度的柚皮苷组,软骨细胞NOS活性均高于空白对照组,低于实验对照组,与空白对照组相比,50ug/mL组有显著性差异(P<0.05)与实验对照组相比,各浓度组柚皮苷NOS活性差异有显著性(P<0.05).结论:骨碎补柚皮苷可促进软骨细胞的增殖,显著提高SOD的活性,降低NOS的活性,从而保护软骨细胞免受肿瘤坏死因子等细胞因子的损伤.     

Expression of inducible nitric oxide synthase in healthy pleura and in malignant mesothelioma

In this study we investigated the immunohistochemical expression of inducible nitric oxide synthase (iNOS) in a set of normal pleural mesothelial tissues, malignant mesotheliomas, mesothelioma cell lines and metastatic pleural adenocarcinomas. Furthermore, the expression of mRNA was assessed in four malignant mesothelioma cell lines in culture. Apoptosis and vascular density in malignant mesotheliomas was assessed by the TUNEL method and by immunohistochemistry with an antibody against FVIII-related antigen. Immunohistochemically mesothelial cells in non-neoplastic healthy pleural tissues were mostly negative for iNOS. Positivity for iNOS was observed in 28/38 (74%) and 24/25 (96%) of malignant mesotheliomas and metastatic pleural adenocarcinomas, respectively. Epithelial and mixed mesotheliomas expressed more often strong iNOS immunoreactivity compared to the sarcomatoid subtype (P = 0.023). Moreover, metastatic adenocarcinomas expressed more often iNOS positivity than mesotheliomas (P = 0.021). Experiments with the cell lines confirmed that malignant mesothelioma cells are capable of synthesizing iNOS. No significant association was found between iNOS expression and apoptosis or vascular density in malignant mesotheliomas. The higher expression of iNOS in the epithelial subtype of mesothelioma and pleural metastatic adenocarcinoma might be due to an increased sensitivity of these cell types to cytokine-mediated iNOS upregulation. The strong expression of iNOS suggests a putative role for NO in the growth and progression of these tumours.     

Nitric oxide synthase 1 is partly compensating for nitric oxide synthase 3 deficiency in nitric oxide synthase 3 knock-out mice and is elevated in murine and human cirrhosis.

BACKGROUND: The role of endothelial nitric oxide synthase 3 (NOS-3) in the hyperdynamic circulation associated with cirrhosis is established but not that of the neuronal (NOS-1) isoform. We therefore investigated aortic NOS-1 levels in NOS-3 knock-out (KO) and wildtype (WT) mice and in hepatic arteries of patients. METHODS: Mice rendered cirrhotic by bile duct ligation (BDL) were compared with sham-operated controls. Hepatic arteries of cirrhotic patients were collected during liver transplantation; donor vessels served as controls. mRNA levels were quantified by real-time PCR, protein levels by Western blotting and NO production by Nomega-nitro-L-arginine methyl ester inhibitable arginine-citrulline assay. RESULTS: Aortae of NOS-3 KO mice exhibited higher NOS-1mRNA (5.6-fold, P < 0.004) and protein levels (8.8-fold) compared with WT. NO production in aortae of NOS-3 KO mice was 52% compared with WT (P = 0.002). BDL increased NOS-1 mRNA (2.4-fold, P = 0.01) and protein (7.1-fold) levels in aortae of WT, but no further in the NOS-3 KO mice. Hepatic artery NOS-1 mRNA levels in cirrhotic patients were markedly increased compared with controls (24.5-fold, P = 0.0007). CONCLUSIONS: Increased NOS-1 mRNA and protein levels and partially maintained in vitro NO-production in aortae of NOS-3 KO mice suggest that NOS-1 may partially compensate for NOS-3 deficiency. BDL-induced increase in aortic NOS-1 mRNA and protein levels hint that not only NOS-3, but also NOS-1 may be involved in the regulation of systemic hyperdynamic circulation and portal hypertension. Upregulation of NOS-1 mRNA levels in hepatic arteries of portal hypertensive patients suggests possible clinical significance for these experimental findings.     

甲基强的松龙对大鼠颅脑损伤后一氧化氮合成酶活性的影响

目的观察甲基强的松龙对大鼠颅脑损伤后血、脑组织中NOS含量的影响,并探讨其作用机制。方法将45只SD大鼠随机分为3组:实验组(20只)、对照组(20只)、正常组(5只),均采用骨窗形成后硬膜外打击法造成鼠脑挫裂伤。正常组麻醉后,只行开颅手术,不作头颅打击,治疗组大鼠致伤后即刻腹腔内注射30mg/kg甲基强的松龙,对照组则注射30mg/kg生理盐水。对照组和治疗组大鼠分别在伤后1h、6h、12h、24h断头取脑,对大鼠脑外伤后脑组织中一氧化碳合成酶(NOS)含量进行检测。结果大鼠脑皮质中的NOS活性在伤后1h较正常组显著性升高(P <0.01),6h开始下降,12～24h降至基础水平。甲基强的松龙治疗组在伤后1h(P <0.01)、6h(P <0.05)NOS活性较损伤组显著性降低。结论颅脑损伤后,受损脑组织中NOS活性升高,甲基强的松龙可通过抑制损伤后NOS活性起到保护创伤神经元的作用。