Chatterjee M, Saluja R, Kanneganti S, Chinta S, Dikshit M
Cardiovascular Pharmacology Unit, Division of Pharmacology, Central Drug Research Institute, Lucknow, India.
Cell Mol Biol (Noisy-le-grand). 2007 Apr 15;53(1):84-93.
Resting neutrophils generate NO, while activation leads to the production of reactive oxygen and nitrogen species. Nowadays cardiovascular pathological conditions such as hypertension, cardiac ischemia, reperfusion and heart failure are associated with inflammation. This project explores the respiratory burst potential and NO generation status in the neutrophils, plasma, aorta, and kidneys from normotensive Wistar and spontaneously hypertensive rats (SHR). Total and protein associated nitrite content was quantitated using Griess reagent following cadmium reduction and mercuric chloride treatment respectively. NO and superoxide generation evaluated by Flowcytometry and peroxynitrite by spectrofluorimetric method. Expression of NOS isoforms was analyzed by RT-PCR. NO generation from SHR neutrophils was significantly augmented in comparison to normotensive counterparts. Neutrophils activated in response to arachidonic acid, PMA, fMLP or E. coli generated more superoxide radicals among SHR, and consequentially peroxynitrite. Expression of iNOS was significantly more in the SHR neutrophils, while that of nNOS remained unaffected. Results suggest that NO generated in SHR is utilized in scavenging superoxide radicals thereby limiting its bioavailability. Thus induction of NOS in neutrophils combined with augmented oxidative stress might influence its association with endothelium and contribute to inflammatory responses under hypertensive condition.
静息中性粒细胞会产生一氧化氮（NO），而激活则会导致活性氧和氮物种的产生。如今，诸如高血压、心脏缺血、再灌注和心力衰竭等心血管病理状况都与炎症相关。本项目探究了正常血压的Wistar大鼠和自发性高血压大鼠（SHR）的中性粒细胞、血浆、主动脉和肾脏中的呼吸爆发潜能以及NO生成状态。分别在镉还原和氯化汞处理后，使用格里斯试剂对总亚硝酸盐含量和蛋白质相关亚硝酸盐含量进行定量。通过流式细胞术评估NO和超氧化物的生成，通过荧光分光光度法评估过氧亚硝酸盐的生成。通过逆转录聚合酶链反应（RT-PCR）分析一氧化氮合酶（NOS）亚型的表达。与正常血压的对应物相比，SHR中性粒细胞产生的NO显著增加。在SHR中，响应花生四烯酸、佛波酯（PMA）、N-甲酰甲硫氨酸-亮氨酸-苯丙氨酸（fMLP）或大肠杆菌而被激活的中性粒细胞会产生更多的超氧自由基，进而产生更多过氧亚硝酸盐。SHR中性粒细胞中诱导型一氧化氮合酶（iNOS）的表达显著更高，而神经元型一氧化氮合酶（nNOS）的表达则未受影响。结果表明，SHR中产生的NO被用于清除超氧自由基，从而限制了其生物利用度。因此，中性粒细胞中NOS的诱导与氧化应激增强相结合，可能会影响其与内皮的关联，并导致高血压状态下的炎症反应。
