大鼠对氟西泮抗痫耐受和依赖时海马中一氧化氮合酶的变化

目的通过测定大鼠对氟西泮抗痫耐受性和依赖性时海马中一氧化氮合酶(NOS)蛋白表达及活性的变化,探讨一氧化氮(NO)在此过程中可能的作用。方法建立大鼠对氟西泮抗痫耐受性和依赖性的模型。运用免疫印迹及免疫组化法检测海马中NOS蛋白表达,以及比色法检测NOS活性的变化。结果大鼠对氟西泮抗痫耐受组的海马中NOS蛋白表达明显高于对照组;而对氟西泮抗痫依赖组则与对照组差别无统计学意义。两组NOS活性均显著高于各自的对照组。结论NO可能是介导氟西泮抗痫耐受性和依赖性的因素之一。     

大鼠脑出血周边组织NO含量和NOS活性变化及对细胞凋亡的影响

目的：研究大鼠脑出血周边组织一氧化氮(NO)含量和一氧化氮合酶(NOS)活性变化及与细胞凋亡的关系。方法：①Wistar大鼠104只,随机分为对照组、脑出血组、脑出血 氮硝基左旋精氨酸(NNLA)组,后两组各分为(4h、6h、12h、1d、3d、7d)6个时间点。②测定出血周边组织NO含量、NOS活性及神经细胞凋亡。结果：①大鼠脑出血周边组织NO、诱导型一氧化氮合酶(iNOS),4h开始升高,3di NOS、NO达峰值。②大鼠脑出血周边组织6h出现凋亡,细胞3d凋亡细胞达峰值,与iNOS峰值对应,7d时仍存在较多凋亡细胞。③NNLA干预后NO含量、iNOS活性及凋亡细胞数量与脑出血组对应时间点比较显著下降,差异显著。结论：大鼠脑出血周边组织神经细胞存在长时间凋亡,NO、iNOS可以促进其凋亡,NOS抑制剂减少大鼠脑出血周边组织神经细胞凋亡。     

荆花胃康胶丸对溃疡大鼠胃黏膜NO,NOS和ET含量的影响

目的:观察荆花胃康胶丸对溃疡大鼠胃黏膜内皮素(ET)、一氧化氮(NO)、一氧化氮合酶(NOS)含量的影响。方法:采用Okabe氏法造成大鼠胃溃疡模型,随机将模型动物分为蒸馏水对照组、荆花胃康胶丸30,20, 10 mg·kg~(-1)·d~(-1)组、硫糖铝10 mg·kg~(-1)·d~(-1)及法莫替丁5 mg·kg~(-1)·d~(-1)组,每组8只。各组均灌胃给药,qd,连续10 d。末次给药后次日,测定各组溃疡面积,并测定溃疡周围组织NO,NOS及ET含量。结果:与对照组相比,荆花胃康胶丸30,20,10 mg·kg~(-1)·d~(-1)组的溃疡面积显著降低,溃疡周围组织NO及NOS的含量明显增高,ET的含量明显降低(P＜0．01),且呈现剂量依赖性;荆花胃康胶丸30 mg·kg~(-1)·d~(-1)组的保护作用优于硫糖铝及法莫替丁组(P＜0．05)。结论:荆花胃康胶丸可能通过增加胃溃疡组织NO及NOS的含量及降低ET的含量来发挥胃黏膜修复作用。     

Involvement of nitric oxide in the inhibition of angiogenesis by interleukin-2

Interleukin-2 (IL-2), an immunoregulatory cytokine possessing antitumour activity, is an inducer of nitric oxide (NO) synthesis in mice and man. In this study, the possibility that IL-2 possesses antiangiogenic properties that account for its antitumour effects in vivo was examined. IL-2 caused a dose-dependent inhibition of angiogenesis in the chick embryo chorioallantoic membrane (CAM). This inhibition was completely reversed by the NO synthase inhibitor N^G-nitro-L-arginine methylester (L-NAME). Furthermore, IL-2 was capable of stimulating NO synthase activity in the CAM in vitro and this effect was suppressed by L-NAME. Addition of IL-2 to human umbilical vein endothelial cells (HUVECs) in culture, had no effect on their growth characteristics. These results suggest that IL-2 may be an important antiangiogenic molecule causing its effect via nitric oxide synthesis. The antiangiogenic activity of IL-2 may be, at least in part, responsible for its antitumour properties.     

四逆汤对失血性休克家兔一氧化氮（NO）和一氧化氮合酶（NOS）的影响

目的：探讨中药四逆汤对失血性休克家兔一氧化氮的影响。方法：采用动物分组对照实验，测量不同状态下动物血浆一氧化氮和一氧化氮合酶(NOS)的变化。结果：与休克前比较，休克组的家兔血浆一氧化氮水平各时段均明显提高(P＜0．01)，而治疗组仅在治疗后30min家兔一氧化氮及NOS水平明显提高(P＜0．01)，其他时段与休克前比较，未见显著差异(P＜0．01)，且休克后lh、4h、8h休克组一氧化氮水平显著高于治疗组(P＜0．01)。经药物治疗的失血性休克家兔血浆一氧化氮水平明显降低。     

鬼针草提取物对实验性高脂血症大鼠血脂和NO及NOS的影响

目的研究鬼针草不同提取物对实验性高脂血症大鼠血脂和NO及NOS的影响。方法健康雄性SD大鼠随机分为5组：①正常对照组;②模型对照组;③洛伐他汀对照组,洛伐他汀2mg/kg·d）灌胃;④鬼针草乙酸乙酯萃取物组,5g／（kg·d）灌胃;⑤鬼针草水提取物组,5g／（kg·d）灌胃,正常组和模型组每日给予等体积生理盐水。除正常组喂饲普通饲料外,其余各组均喂饲高脂饲料。10周末处死动物,分离血清,检测血脂、MDA、SOD、NO、NOS。制备腹主动脉切片,HE染色观察形态学改变。结果与模型组比较,鬼针草提取物组TC、LDL—C、MDA、NO和NOS显著降低,HDL—C、SOD明显升高。鬼针草提取物组HDL-C明显高于洛伐他汀组。模型组大鼠主动脉血管内壁不光滑,血管内膜脂质沉积,鬼针草提取物组血管内膜无明显的脂质沉积。结论鬼针草提取物能够降低高脂血症大鼠血脂水平并保护血管内皮,具有较好的防治动脉粥样硬化的作用。     

芍药苷对缺氧损伤内皮细胞产生NO、eNOS和细胞粘附分子的影响

目的:观察芍药苷对缺氧损伤的人脐静脉内皮细胞(HUVEC)产生一氧化氮(NO)、内皮型一氧化氮合酶(eNOS)和细胞粘附分子(ICAM-1、VCAM-1)的影响。方法:体外培养HUVEC,传至3代后,以不同浓度的芍药苷分别作用于HUVEC,同时进行缺氧处理。以硝酸还原酶法测定培养液上清中的NO,免疫细胞化学法检测内皮细胞eNOS的表达,细胞ELISA法测定细胞表面ICAM-1和VCAM-1的含量。结果:HUVEC在缺氧48h后产生NO的量显著减少(P<0.001),eNOS表达下调,而ICAM-1、VCAM-1表达上调;芍药苷可以剂量依赖性的增加内皮细胞NO生成量,上调eNOS的表达,下调ICAM-1、VCAM-1表达。结论:芍药苷可能通过增加HUVEC eNOS的表达增加NO的释放、抑制ICAM-1及VCAM-1的表达等途径对内皮细胞起保护作用。     

硒和/或维生素E对实验性高脂血症大鼠心、肝、肾及血清中一氧化氮及一氧化氮合酶的影响

目的 :观察硒和/或维生素E(VE)对实验性高脂血症大鼠心、肝、肾、血清一氧化氮 (NO)及一氧化氮合酶 (NOS)的影响。方法 :对大鼠喂以高脂饲料致实验性高脂血症 ,然后分别分组给予硒和/或VE ,4wk后取血及心、肝、肾组织匀浆 ,采用硝酸还原酶等方法测定上述组织的NO和NOS含量。结果 :高脂饲料可致血清、心、肝、肾组织中NO含量及NOS活性降低 ;硒和/或VE能不同程度地增加这些组织中NO含量及心、肝、肾中的NOS活性 (P<0 05或P<0 01) ,且两者合用比单用作用更明显。结论 :硒和/或VE可致实验性高脂血症大鼠心、肝、肾及血清中的NO和NOS发生改变。     

复荣通脉胶囊对糖尿病大鼠血脂代谢及eNOS的影响

目的:探讨复荣通脉胶囊对糖尿病大鼠血脂及内皮型一氧化氮合酶(eNOS)表达的影响。方法:采用腹腔注射链脲佐菌素法建立糖尿病大鼠模型,按完全随机原则分为正常组、模型组与复荣通脉胶囊低剂量治疗(FRL)组、中剂量治疗(FRM)组、高剂量治疗(FRH)组。连续治疗8周后,腹主动脉采血检测胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C);免疫组化法检测心肌eNOS的表达。结果:造模各组与正常组相比,TG水平明显升高(P<0.01),TC、LDL-C、HDL-C差异无统计学意义(P>0.05);复荣通脉胶囊各治疗组与糖尿病模型组比较,TC、TG、LDL-C、HDL-C水平差异无统计学意义(P>0.05),复荣通脉胶囊各治疗组间TC、TG、LDL-C、HDL-C水平差异无统计学意义(P>0.05)。造模各组与正常组相比,心肌eNOS表达明显降低(P<0.01);FRL组与模型组比较,心肌eNOS表达差异无统计学意义(P>0.05);FRM组、FRH组与模型组、FRL组相比,心肌eNOS均明显增高(P<0.01),但FRM组与FRH组比较,差异无统计学意义(P>0.05)。结论:复荣通脉胶囊并非通过纠正糖脂代谢、减轻体质量起作用,而是通过减轻内皮细胞损伤与改善心肌微血管内皮细胞功能,以减少糖尿病心肌病的发生与发展。     

Inhibitory Effects of Ginsenoside-Rb2 on Nicotinic Stimulation-Evoked Catecholamine Secretion

The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 (3~30 µM), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 (10 µM) also time-dependently inhibited the CA secretion evoked by DMPP (100 µM, a selective neuronal nicotinic receptor agonist) and high K⁺ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 (50 µg/mL), the secretory responses of CA evoked by veratridine (a selective Na⁺ channel activator (50 µM), Bay-K-8644 (an L-type dihydropyridine Ca²⁺ channel activator, 10 µM), and cyclopiazonic acid (a cytoplasmic Ca²⁺-ATPase inhibitor, 10 µM) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 (10 µM) and L-NAME (an inhibitor of NO synthase, 30 µM), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 (10 µM) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of Ca²⁺ and Na⁺ into the adrenomedullary chromaffin cells and also by suppressing the release of Ca²⁺ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.     

Drug Discovery for Overcoming Chronic Kidney Disease (CKD): The Endothelin ETB Receptor / Nitric Oxide System Functions as a Protective Factor in CKD

Accelerated cardiovascular disease (CVD) is a frequent complication of renal disease. Chronic kidney disease (CKD) develops hypertension and dyslipidemia, which in turn can contribute to the progression of renal failure. There is general agreement that endothelin-1 (ET-1), which acts through the two subtypes of receptor ET_A and ET_B, plays important physiological roles in the regulation of normal cardiovascular function and that excessive ET-1 production is linked to CVD and CKD. Although selective ET_A or nonselective ET_A/ET_B receptor antagonisms have been recognized as a potential strategy for treatment of several cardiovascular disease, it remains unclear which of the antagonisms is suitable for the individuals with CKD because upregulation of the nitric oxide (NO) system via ET_B receptor is responsible for renal function such as natriuresis, diuresis, and glomerular hemodynamics. Our findings clearly indicate that the blockade of ET receptors, in particular ET_A-receptor antagonism, not only produces a potential renoprotective effect in CKD but also reduces the risk of CVD. In contrast, pharmacological blockade or genetic deficiency of ET_B receptor seems to aggravate CKD and CVD in several experimental models of rats. Moreover, preliminary evidence in patients with CKD also suggests that both selective ET_A- and nonselective ET_A/ET_B-receptor blockade decreases blood pressure but that selective ET_A blockade has additional desirable effects on renal hemodynamics. Thus, at least in CKD, these findings support the notion that ET_B receptor– mediated actions produce a renoprotective effect and that nonselective ET_A/ET_B-receptors blockade seem to offer no advantage over selective ET_A antagonism, and if anything may potentially reduce the benefits.     

Cigarette smoke-inhibition of neurogenic bronchoconstriction in guinea-pigs in vivo: involvement of exogenous and endogenous nitric oxide

1. We investigated the effect of acute inhalation of cigarette smoke on subsequent non-adrenergic, non-cholinergic (NANC) neural bronchoconstriction in anaesthetized guinea-pigs in vivo by use of pulmonary insufflation pressure (PIP) as an index of airway tone. The contribution of endogenous nitric oxide (NO) was investigated with the NO synthase inhibitor N^G-nitro-L-arginine methyl ester (L-NAME). The contribution of plasma exudation to the response was investigated with Evans blue dye as a plasma marker.
2. Inhalation of 50 tidal volumes of cigarette smoke or air had no significant effect on baseline PIP. In the presence of propranolol and atropine (1 mg kg⁻¹ each), electrical stimulation of the vagus nerves in animals given air 30 min previously induced a frequency-dependent increase in PIP above sham stimulated controls (16 fold increase at 2.5 Hz, 24 fold increase at 10 Hz). In contrast, in smoke-exposed animals, the increase in subsequent vagally-induced PIP was markedly less than in the air controls (90% less at 2.5 Hz, 76% less at 10 Hz).
3. L-NAME (10 mg kg⁻¹), given 10 min before air or smoke, potentiated subsequent vagally-induced (2.5 Hz) NANC bronchoconstriction by 338% in smoke-exposed animals, but had no significant effect in air-exposed animals. The inactive enantiomer D-NAME (10 mg kg⁻¹) had no effect, and the potentiation by L-NAME was partially reversed by the NO-precursor L-arginine (100 mg kg⁻¹). Vagal stimulation did not affect the magnitude of vagally-induced bronchoconstriction 30 min later.
4. Cigarette smoke exposure reduced the magnitude of subsequent bronchoconstriction induced by neurokinin A (NKA) by 37% compared with the effect of NKA in air-exposed animals. L-NAME had no significant effect on the smoke-induced inhibition of NKA-induced bronchoconstriction.
5. Vagally-induced plasma exudation in the main bronchi was greater in smoke-exposed animals compared with air-exposed animals (120% greater at 2.5 Hz, 82% greater at 10 Hz).
6. We conclude that cigarette smoke-induced inhibition of subsequent NANC neurogenic bronchoconstriction is not associated with inhibition of airway plasma exudation and is mediated in part via exogenous smoke-derived NO, or another bronchoprotective molecule, and by endogenous NO.

     

前列地尔对局灶性脑缺血模型大鼠大脑皮层中iNOS和eNOS表达的影响

目的:探讨前列地尔对局灶性脑缺血模型大鼠大脑皮层中诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)表达的影响。方法:取大鼠随机分为假手术组、模型组、阳性对照(尼莫地平1mg.kg-1)组和前列地尔低、中、高剂量(1.25、2.50、5.00μg.kg-1)组,每组10只,腹腔注射相应药物6d,每日1次,末次给药后建立局灶性脑缺血模型,大鼠清醒后进行神经功能缺损评分,采用免疫组化法和免疫印迹法检测各组大鼠建模2h后iNOS、eNOS的表达。结果:与假手术组比较,其余各组神经功能缺损评分均明显增加,iNOS阳性细胞数和表达量均明显增加,除模型组外其余各组eNOS阳性表达量均增加(P均<0.05);与模型组比较,阳性对照组和前列地尔中、高剂量组神经功能缺损评分均明显降低,阳性对照组和前列地尔3个剂量组iNOS阳性细胞数和表达量均明显降低、eNOS阳性细胞数和表达量均明显增加(P均<0.05)。结论:前列地尔预处理对局灶性脑缺血模型大鼠具有保护作用,其机制可能与下调大脑皮层iNOS、上调eNOS的表达有关。     

Effects of Nicorandil on the cAMP-Dependent Cl− Current in Guinea-Pig Ventricular Cells

In guinea-pig cardiomyocytes, a cAMP-dependent Cl^? current (I_Cl,cAMP) flows through a cardiac isoform of the cystic fibrosis transmembrane conductance regulator (CFTR), which belongs to a family of the ATP-binding cassette (ABC) proteins. Although several K⁺-channel openers and sulfonylurea ATP-sensitive K⁺ (K_ATP)–channel blockers reportedly inhibit I_Cl,cAMP, effects of nicorandil on the Cl^? current have not been evaluated. This study was conducted to examine the effects of nicorandil on I_Cl,cAMP in isolated guinea-pig ventricular cells using patch clamp techniques. Nicorandil in concentrations higher than 300 μM enhanced the I_Cl,cAMP preactivated by 0.1 μM isoproterenol. The isoproterenol-induced I_Cl,cAMP was inhibited by 100 μM glibenclamide, but not by 100 μM pinacidil. SNAP (S-nitroso-N-acetyl-d,l-penicillamine, 10 μM), a nitric oxide (NO) donor, similarly enhanced the isoproterenol-induced I_Cl,cAMP. However, SG-86, a denitrated metabolite possessing K⁺ channel–opening action, failed to enhance the Cl^? current. When the I_Cl,cAMP was activated by 3-isobutyl-1-methylxanthine (IBMX, 30 μM), either nicorandil or SNAP failed to enhance the isoproterenol-induced I_Cl,cAMP. Thus, nicorandil enhances I_Cl,cAMP in guinea-pig cardiomyocytes through an increase in intracellular cGMP, although direct modulation of I_Cl,cAMP by NO cannot be completely excluded.     

杏仁核中一氧化氮对睡眠-觉醒的影响

目的研究杏仁核中一氧化氮（ＮＯ）对大鼠睡眠 觉醒的影响,并分析其作用机制。方法多导睡眠描记和杏仁核微量注射。结果一氧化氮合酶抑制剂Ｌ 硝基精氨酸（Ｌ ＮＮＡ）可增加慢波睡眠（ＳＷＳ）和减少觉醒（Ｗ）,而一氧化氮（ＮＯ）供体硝普钠（ＳＮＰ）可增加Ｗ、减少ＳＷＳ,并可对抗Ｌ ＮＮＡ的促睡眠效应;ＮＯ前体Ｌ 精氨酸（Ｌ Ａｒｇ）对睡眠 觉醒无直接影响,但可对抗Ｌ ＮＮＡ的促睡眠效应。环磷酸鸟苷（ｃＧＭＰ）具有明显的增加Ｗ和减少ＳＷＳ效应,而鸟苷酸环化酶抑制剂亚甲蓝（ＭＢ）增加睡眠、减少觉醒,并可阻断ＳＮＰ的促睡眠效应。结论杏仁核参与睡眠 觉醒调节,杏仁核中ＮＯ具有促进Ｗ、抑制ＳＷＳ效应,这一作用是通过激活鸟苷酸环化酶使ｃＧＭＰ增多实现的。     

Challenges and opportunities with modelling and simulation in drug discovery and drug development

The benefits of modelling and simulation at the pre-clinical stage of drug development can be realized through formal and realistic integration of data on physicochemical properties, pharmacokinetics, pharmacodynamics, formulation and safety. Such data integration and the powerful combination of physiologically based pharmacokinetic (PBPK) with pharmacokinetic–pharmacodynamic relationship (PK/PD) models provides the basis for quantitative outputs allowing comparisons across compounds and resulting in improved decision-making during the selection process. Such PBPK/PD evaluations provide crucial information on the potency and safety of drug candidates in vivo and the bridging of the PK/PD concept established during the pre-clinical phase to clinical studies. Modelling and simulation is required to address a number of key questions at the various stages of the drug-discovery and -development process. Such questions include the following. (1) What is the expected human PK profile for potential clinical candidate(s)? (2) Is this profile and its associated PD adequate for the given indication? (3) What is the optimal dosing schedule with respect to safety and efficacy? (4) Is a food effect expected? (5) How can formulation be improved and what is the potential benefit? (6) What is the expected variability and uncertainty in the predictions?     

重组人生长激素对败血症大鼠急性肺损伤的治疗及其机制

目的　观察重组人生长激素(rhGH)对败血症大鼠急性肺损伤(ALI)的治疗效果并探讨其作用机制。方法　采用鸵鸟株E .coli复制败血症大鼠ALI模型。观测对照组、ALI组及rhGH治疗组大鼠的肺组织病理评分、支气管肺泡灌洗液(BALF)中NO水平及肺组织诱生型一氧化氮合酶(iNOS)蛋白的表达情况。结果　ALI组大鼠肺组织病理评分、BALF中NO水平及肺组织iNOS蛋白表达水平明显高于对照组,且NO与iNOS蛋白表达水平呈明显正相关。rhGH能明显降低败血症大鼠BALF中NO及肺组织iNOS水平,减轻肺损伤程度。结论　由iNOS诱导产生的过量NO可促进败血症大鼠ALI的发生;rhGH对败血症大鼠ALI具有较理想的治疗效果,其机制可能与rhGH通过影响iNOS表达来降低肺组织NO水平等有关。     

复方花刺参黏多糖对球囊血管成形术兔血浆内皮素及血清一氧化氮的影响

目的 探讨复方花剌参黏多糖对家兔髂动脉腔内成形术(TA)后血浆内皮素及血清一氧化氮的影响。方法 50只新西兰家兔随机分为4组：花剌参组、辛伐他汀组、模型组和正常组。花刺参组、辛伐他汀组和模型组用球囊导管剥脱损伤髂动脉内皮后。喂高脂饲料6wk，髂动脉造影显示形成粥样硬化狭窄．行TA后，即日均停喂高脂饲料改普通饲料，花刺参组和辛伐他汀组经胃管给药，模型组给等量生理盐水，分笼喂养，自由饮水。正常组仅给予普通饮食及假手术。各组动物于TA后4wk空腹颈动脉同步采血行血浆内皮素、血清一氧化氮浓度测定。结果 TA后4wk，模型组与正常组相比血浆ET浓度明显升高。而血清NO浓度明显降低，花刺参组和辛伐他汀组血浆ET浓度显著低于模型组．而血清NO浓度显著高于模型组。结论 家兔髂动脉TA后ET增多，NO减少，存在血管内皮功能失调，而复方花刺参黏多糖通过调整ET和NO的平衡改善血管内皮功能。     

当归多糖对AD模型小鼠抗衰老作用研究

目的观察当归多糖对AD模型小鼠学习记忆功能,测定脑组织中NO含量、p16蛋白含量、NOS及AchE活性,探讨当归多糖治疗衰老的作用机制。方法 KM小鼠72只,采用Y型迷宫实验测定小鼠的学习记忆能力,应用生化法测定脑组织NO含量、NOS及AchE活性,ELISA法测定p16蛋白含量。结果当归多糖各剂量组小鼠学习记忆能力明显得到改善,脑组织中NO浓度、NOS活性、AchE活性和p16蛋白含量与对照组比较显著降低。结论当归多糖具有延缓衰老的作用,可能机制是通过抗氧化作用和调控细胞周期实现的。     

弥漫性颅脑损伤对大鼠海马一氧化氮合成酶活性及表达的影响

目的 :研究大鼠弥漫性颅脑损伤后海马结构一氧化氮合成酶 (NOS)活力及表达 ,分析海马不同亚区NOS活力变化与时间的关系。方法 :制作Wistar大鼠弥漫性脑损伤模型 ,并分对照组、假手术组及伤后6、12、24、48h组 ,于不同时间点获取脑组织 ;用NADPH 黄递酶 (NADPH d)组织化学法和免疫组化法检测NOS的活性及表达情况。结果 :NADPH d组织化学法显示 ,弥漫性脑损伤后 ,海马结构NOS阳性神经元数量在伤后6h最多 ,以后逐渐减少 ,伤后24、48h组明显低于假手术组 (P<0 05) ;免疫组化结果显示 ,在CA1、CA3区和齿状回 (DG) ,伤后24、48h组阳性细胞数均低于假手术组 (P<0 01)。结论 :大鼠弥漫性脑损伤后 ,可引起海马各区NOS的变化 ,该变化可能是造成脑组织继发性损害机制之一。