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中性粒细胞明胶酶相关载脂蛋白通过凋亡调节蛋白保护缺血/再灌注损伤大鼠肾小管上皮细胞。

Neutrophil gelatinase-associated lipocalin protects renal tubular epithelial cell in ischemic/reperfusion injury rats via apoptosis-regulating proteins.

机构信息

Department of Nephrology, Shanghai First People's Hospital Affiliated to Jiaotong University, Shanghai, PR China.

出版信息

Ren Fail. 2012;34(6):777-83. doi: 10.3109/0886022X.2012.678173. Epub 2012 Apr 16.

DOI:10.3109/0886022X.2012.678173
PMID:22500534
Abstract

We investigated the role of neutrophil gelatinase-associated lipocalin (NGAL) on renal tubular epithelial cell in the renal ischemia/reperfusion injury (IRI) rats. Male Sprague-Dawley rats were randomly assigned to three groups. The control group (n = 5) underwent left nephrectomy. The ischemia/reperfusion (I/R) + normal saline (NS) (n = 5) and I/R + NGAL groups (n = 5) were subjected to 45 min right renal ischemia followed by 48 h of reperfusion after left nephrectomy. The pathological changes of kidney tissues were investigated using hematoxylin-eosin staining; renal tubular epithelial cell apoptosis was detected using terminal dUTP nick-labeling method; expression of apoptosis-regulating protein Fas and Bcl-2 was measured using real-time polymerase chain reaction, Western blot, and immunohistochemical staining. Compared with I/R + NS group, kidney tissues from I/R + NGAL group revealed reduced histological damage and a decreased number of renal tubular epithelial cell apoptosis (9.2 ± 2.53 nuclei or 4.0 ± 0.7 per high-power field vs. 20.3 ± 3.7 nuclei or 8.1 ± 0.3 per high-power field); rats with NGAL showed downregulated fas mRNA (2.34 ± 0.51 vs. 6.84 ± 2.34), fas protein (0.65 ± 0.05 vs. 0.95 ± 0.08), and upregulated bcl-2 protein (0.33 ± 0.05 vs. 0.24 ± 0.03). The results had statistical significance (p < 0.05). We think NGAL could protect against renal IRI and might be related to decreasing tubular epithelial cell apoptosis via adjusting the expression of apoptosis-regulating proteins.

摘要

我们研究了中性粒细胞明胶酶相关脂质运载蛋白(NGAL)在肾缺血/再灌注损伤(IRI)大鼠肾小管上皮细胞中的作用。雄性 Sprague-Dawley 大鼠随机分为三组。对照组(n = 5)行左肾切除术。缺血/再灌注(I / R)+生理盐水(NS)组(n = 5)和 I / R + NGAL 组(n = 5)在左肾切除后 45 分钟行右肾缺血,再灌注 48 小时。苏木精-伊红染色法观察肾组织病理变化;末端脱氧核苷酸转移酶介导的缺口末端标记法检测肾小管上皮细胞凋亡;实时聚合酶链反应、Western blot 和免疫组织化学染色法检测凋亡调节蛋白 Fas 和 Bcl-2 的表达。与 I / R + NS 组相比,I / R + NGAL 组肾组织损伤减轻,肾小管上皮细胞凋亡减少(9.2 ± 2.53 个核或 4.0 ± 0.7 个/高倍视野比 20.3 ± 3.7 个核或 8.1 ± 0.3 个/高倍视野);NGAL 组 fas mRNA(2.34 ± 0.51 比 6.84 ± 2.34)、fas 蛋白(0.65 ± 0.05 比 0.95 ± 0.08)下调,bcl-2 蛋白(0.33 ± 0.05 比 0.24 ± 0.03)上调,差异均有统计学意义(p < 0.05)。我们认为 NGAL 可能通过调节凋亡调节蛋白的表达来保护肾脏免受 IRI,可能与减少肾小管上皮细胞凋亡有关。

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