Shokeir A A, Barakat N, Hussein A M, Awadalla A, Harraz A M, Khater S, Hemmaid K, Kamal A I
Urology and Nephrology Center, Mansoura University and Physiology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
Physiol Res. 2015;64(3):313-23. doi: 10.33549/physiolres.932834. Epub 2014 Dec 22.
Objectives of the study were to investigate impact of ischemic preconditioning (Ipre) and sulforaphane (SFN) and combination of them on nuclear factor 2 erythroid related factor 2 (Nrf2) gene and its dependent genes, heme oxygenase-1 (HO1) and NADPH-quinone oxidoreductase1 (NQO-1) and inflammatory cytokines TNF-alpha, IL1beta, and intercellular adhesion molecule-1 (ICAM1) and caspase-3 in renal ischemia/reperfusion (I/R) injury. Ninety male Sprague Dawely rats were classified into 5 groups (each consists of 18 rats): sham, control, Ipre, sulforaphane and Sulfo+Ipre. Each group was subdivided into 3 subgroups each containing 6 rats according to time of harvesting kidney and taking blood samples; 24 h, 48 h, and 7 days subgroups. Renal functions including serum creatinine, BUN were measured at basal conditions and by the end of experiment. Expression of Nrf2, HO-1, NQO-1, TNF-alpha, IL-1beta, and ICAM-1 was measured by real time PCR in kidney tissues by the end of experiment. Also, immunohistochemical localization of caspase-3 and chemical assay of malondialdehyde (MDA), GSH and SOD activity were measured in kidney tissues. Both Ipre and SFN improved kidney functions, enhanced the expression of Nrf2, HO-1, and NQO-1, attenuated the expression of inflammatory (TNF-alpha, IL-1, and ICAM-1) and apoptotic (caspase-3) markers. However, the effect of sulforaphane was more powerful than Ipre. Also, a combination of them caused more improvement in antioxidant genes expression and more attenuation in inflammatory genes but not caspase-3 than each one did separately. Sulforaphane showed more powerful effect in renoprotection against I/R injury than Ipre as well as there might be a synergism between them at the molecular but not at the function level.
本研究的目的是探讨缺血预处理（Ipre）、萝卜硫素（SFN）及其联合应用对核因子2红细胞相关因子2（Nrf2）基因及其依赖基因血红素加氧酶-1（HO1）和NADPH-醌氧化还原酶1（NQO-1）以及炎性细胞因子肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL1β）和细胞间黏附分子-1（ICAM1）及半胱天冬酶-3在肾缺血/再灌注（I/R）损伤中的影响。90只雄性Sprague Dawely大鼠被分为5组（每组18只）：假手术组、对照组、Ipre组、萝卜硫素组和萝卜硫素+Ipre组。根据取肾和采血时间，每组再分为3个亚组，每组6只大鼠；分别为24小时亚组、48小时亚组和7天亚组。在基础状态和实验结束时测量包括血清肌酐、血尿素氮在内的肾功能。实验结束时，通过实时聚合酶链反应（PCR）检测肾组织中Nrf2、HO-1、NQO-1、TNF-α、IL-1β和ICAM-1的表达。此外，还检测了肾组织中半胱天冬酶-3的免疫组化定位以及丙二醛（MDA）、谷胱甘肽（GSH）和超氧化物歧化酶（SOD）活性的化学分析。Ipre和SFN均改善了肾功能，增强了Nrf2、HO-1和NQO-1的表达，减弱了炎性（TNF-α、IL-1和ICAM-1）和凋亡（半胱天冬酶-3）标志物的表达。然而，萝卜硫素的作用比Ipre更强。此外，与单独使用相比，二者联合应用对抗氧化基因表达的改善作用更大，对炎性基因的减弱作用更强，但对半胱天冬酶-3无此作用。萝卜硫素在肾保护方面对I/R损伤的作用比Ipre更强，并且它们在分子水平上可能存在协同作用，但在功能水平上不存在协同作用。
