抗黏附治疗对大鼠急性肾损伤中树突状细胞迁移聚集的影响

Effect of Anti-adhesive Treatment on Dendritic Cell Immigration and Accumulation in Rat Renal Ischemia-reperfasion Injury

目的 探讨黏附分子P-选择素、ICAM-1及树突状细胞(DC)在大鼠肾缺血再灌注损伤中作用,以及抗P-选择素单抗的抗黏附抑制及其防治效果。方法 建立肾缺血再灌注损伤大鼠模型,随机分为P-选择素单抗治疗组(n=20)和非治疗组(n=20)。按不同再灌注时间(1,3,6和24h)再分为4组,另设假手术组(n=5)作为对照。采用免疫组化LSAB法和免疫双染与荧光图像分析法,分别观察大鼠肾组织中P-选择素、ICAM-1表达及CD1a^+CD80^+DC分布变化;同时观察丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性以及细胞凋亡等改变。结果 ①缺血再灌注1h起,P-选择素即以肾小管上皮细胞为主于肾内广泛高表达,至24h仍维持一定量水平;ICAM-1于缺血再灌注6h起持续以肾血管为主表达上调和明显增强。②CD1a^+CD80^+DC自再灌注1h起以肾小管间质为主分布数量逐渐增多,至24h达峰,并与大鼠血尿素氮和肌酐水平密切相关。③此外缺血再灌注后,大鼠肾组织MDA含量增加,SOD活性下降,出现明显的肾组织细胞凋亡。④经抗P-选择素单抗处理后,大鼠肾组织P-选择素和ICAM-1表达受到抑制,CD1a^+CD80^+DC分布及数量减少,MDA含量降低和SOD活性上升,细胞凋亡减少,以及肾组织病理损伤和肾功能也相应减轻和改善。结论 本研究提示。

Objective To investigate the role of P-selectin,intercellular adhesive molecule-1(ICAM -1) and dendritic cells(DCs) in rat kidney with renal ischemia-reperfusion injury, as well as the preventive effect of anti-P-seleclin lectin-ECF domain monoclonal antibody (PsL-ECFmAb) .Methods Rat models of renal ischemia - reperfusion were established, which were divided into treated group with PsL - EGF mAb( n = 20) and untreated group(n = 20), which included 4 groups repectively according to reperfusion time (1, 3, 6, 24h) ,sham group(n = 5)served as control. DCs were observed by microscopy image method, while P-selectin and ICAM - 1 were analysed by immunohistochemistry LSAB, and MDA contents, SOD activities and cell apoptosis were detected at the same time. Results (1)P-selectin presented was increased evidently in renal tubular epithelial cell alter ischemia-reperfusion 1h, and the expression of ICAM - 1 was up - regulated in renal vessels after 6h.(2)CD1a+CD80+DC was gradually found in renal tubules and interstitium from ischemia - reperfusion 1h , and number of DCs of 24h group is greater than others. The localization of DCs was associated with rat blood uria nitrogen and serum creatinine(P < 0.05) .(3)Increased MDA contents, declined SOD activities and cell apoptosis were found in renal tissues following rat renal ischemia-reperfusion injury.(4)These changes became less conspicuous in rat treated with PsL - EGFmAb. Conclusions DCs play an important role in immune pathogenesis of renal ischemia-reperiusion injury. PsL - ECFmAb may adjust and inhibit local DCs immigration and accumulation of kidney.