Duan Guo-xian, Zhao Li-jun, Zhang Lian-yuan, Dong Shu-yun, Zhou Hong-xia, Zhang Na, Wang Yin-huan, Men Xiu-li, Li Hong-jie
Department of Pathophysiology of North China Coal Medical College, Tangshan 063000, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2007 Nov;23(4):456-61.
To probe into the affection and significance of NO on the expression of P-selectin in renal injury following hind limb ischemia/reperfusion in rats.
In accordance with the conventional approaches of our department, the model rats were prepared after they were made to undergo 4 hours or ischemia followed by 4 hours of reperfusion of hind limbs. The Wistar rats were divided into four groups randomly: Control group, LI/R group, L-Arg group and L-NAME group. And then in those four groups of Wistar rats, a series of values of measurement were determined such as: Plasma concentrations of nitric oxide (NO), blood urea nitrogen (BUN) and creatinine (Cr). Furthermore, biochemically there came to the assessment of the values including myeloperoxidase (MPO), NO, total nitric oxide synthase (tNOS), inducible NOS (iNOS) and constitutive NOS (cNOS) of renal tissue in different groups. By the methods of electrophoresis and biochemistry, the urine protein was mensurated. The immunohistochemical method was used to detect the expression of P-selectin protein. The morphologic changes were observed with a microscope.
After hind limbs had suffered from ischemia/reperfusion for 4 hours, there was the occurrence of a series of results such as in the following which were based on the comparison between plasm of LI/R group and control group. The values of NO, BUN and Cr increased significantly, and the trend of indexes such as NO in renal tissue was similar to that in plasma. The values of MPO, tNOS and iNOS in renal tissue all increased significantly after reperfusion, while cNOS decreased distinctly. The urine protein appeared, especially large molecular weight protein. Renal pathology revealed that after LI/R there were edema and infiltration of polymorphonuclear neutrophil (PMN). Immunohistochemically, the expression of P-selectin was upregulated significantly. Compared with LI/R rats, all injury changes were alleviated in L-Arg group. Morphologic changes were mild. Both the content of urine protein and the percentage of apoptosis cell decreased. The expression of P-selectin was downregulated. In L-NAME group, all injury changes got worse. Immunohistochemical results showed strong positive staining of P-selectin.
The renal injury after LI/R may relate to the strong expression of P-selectin. NO may have protective affection by decreasing the expression of P-selectin and alleviating the adhesion, aggregation and infiltration of neutrophils.
探讨一氧化氮（NO）对大鼠后肢缺血/再灌注肾损伤中P-选择素表达的影响及其意义。
按照本科室常规方法，制备后肢缺血4小时再灌注4小时的模型大鼠。将Wistar大鼠随机分为四组：对照组、缺血/再灌注组（LI/R组）、L-精氨酸组（L-Arg组）和L-硝基精氨酸甲酯组（L-NAME组）。然后测定四组Wistar大鼠的一系列测量值，如血浆一氧化氮（NO）、血尿素氮（BUN）和肌酐（Cr）浓度。此外，对不同组肾组织的髓过氧化物酶（MPO）、NO、总一氧化氮合酶（tNOS）、诱导型一氧化氮合酶（iNOS）和组成型一氧化氮合酶（cNOS）的值进行生化评估。采用电泳和生化方法测定尿蛋白。用免疫组织化学方法检测P-选择素蛋白的表达。用显微镜观察形态学变化。
后肢缺血/再灌注4小时后，基于LI/R组与对照组血浆比较出现了一系列结果。NO、BUN和Cr值显著升高，肾组织中NO等指标的变化趋势与血浆相似。再灌注后肾组织中MPO、tNOS和iNOS值均显著升高，而cNOS明显降低。出现尿蛋白，尤其是大分子蛋白。肾脏病理学显示，缺血/再灌注后有水肿和多形核中性粒细胞（PMN）浸润。免疫组织化学显示，P-选择素的表达显著上调。与LI/R大鼠相比，L-Arg组所有损伤变化均减轻。形态学变化轻微。尿蛋白含量和凋亡细胞百分比均降低。P-选择素的表达下调。在L-NAME组，所有损伤变化均加重。免疫组织化学结果显示P-选择素呈强阳性染色。
缺血/再灌注后肾损伤可能与P-选择素的强表达有关。NO可能通过降低P-选择素的表达，减轻中性粒细胞的黏附、聚集和浸润而具有保护作用。
