坏死性凋亡在大鼠 CKD 模型对比剂诱导肾病中的作用。

Involvement of necroptosis in contrast-induced nephropathy in a rat CKD model.

机构信息

Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, S-1, W-16, Chuo-ku, Sapporo, 060-8543, Japan.

出版信息

Clin Exp Nephrol. 2021 Jul;25(7):708-717. doi: 10.1007/s10157-021-02048-1. Epub 2021 Mar 16.

DOI:10.1007/s10157-021-02048-1

PMID:33728555

Abstract

BACKGROUND

The risk of contrast-induced nephropathy (CIN) is high in patients with chronic kidney disease (CKD). However, the mechanism of CIN in CKD is not fully understood. Here, we prepared a clinically relevant model of CIN and examined the role of necroptosis, which potentially cross-talks with autophagy, in CIN.

METHODS

In Sprague-Dawley rats, CKD was induced by subtotal nephrectomy (SNx, 5/6 nephrectomy) 4 weeks before induction of CIN. CIN was induced by administration of a contrast medium (CM), iohexol, following administration of indomethacin and N-omega-Nitro-L-arginine methyl ester. Renal function and tissue injuries were assessed 48 h after CM injection.

RESULTS

Serum creatinine (s-Cre) and BUN were increased from 0.28 ± 0.01 to 0.52 ± 0.02 mg/dl and from 15.1 ± 0.7 to 29.2 ± 1.2 mg/dl, respectively, after SNx alone. CM further increased s-Cre and BUN to 0.69 ± 0.03 and 37.2 ± 2.1, respectively. In the renal tissue after CM injection, protein levels of receptor-interacting serine/threonine-protein kinase (RIP) 1, RIP3, cleaved caspase 3, and caspase 8 were increased by 64 ~ 212%, while there was reduction in LC3-II and accumulation of p62. Necrostatin-1, an RIP1 inhibitor, administered before and 24 h after CM injection significantly suppressed elevation of s-Cre, BUN and urinary albumin levels, kidney injury molecule-1 expression and infiltration of CD68-positive macrophages in renal tissues after CM injection.

CONCLUSION

The results suggest that necroptosis of proximal tubular cells contributes to CIN in CKD and that suppression of protective autophagy by pro-necroptotic signaling may also be involved.

摘要

背景

患有慢性肾病（CKD）的患者发生造影剂诱导的肾病（CIN）的风险很高。然而，CKD 中 CIN 的机制尚未完全阐明。在这里，我们制备了一种临床相关的 CIN 模型，并研究了潜在与自噬相互作用的坏死性凋亡在 CIN 中的作用。

方法

在 Sprague-Dawley 大鼠中，在诱导 CIN 前 4 周通过 5/6 肾切除术（SNx）诱导 CKD。在给予吲哚美辛和 Nω-硝基-L-精氨酸甲酯后，给予造影剂（碘海醇）诱导 CIN。在 CM 注射后 48 小时评估肾功能和组织损伤。

结果

SNx 单独治疗后，血清肌酐（s-Cre）和 BUN 从 0.28±0.01mg/dl 增加到 0.52±0.02mg/dl 和 15.1±0.7mg/dl 增加到 29.2±1.2mg/dl。CM 进一步将 s-Cre 和 BUN 增加到 0.69±0.03mg/dl 和 37.2±2.1mg/dl。在 CM 注射后的肾组织中，受体相互作用丝氨酸/苏氨酸蛋白激酶（RIP）1、RIP3、裂解的半胱天冬酶 3 和半胱天冬酶 8 的蛋白水平增加了 64%至 212%，而 LC3-II 减少，p62 积累。在 CM 注射前和注射后 24 小时给予 RIP1 抑制剂 Necrostatin-1，可显著抑制 CM 注射后 s-Cre、BUN 和尿白蛋白水平的升高，肾损伤分子-1 表达和 CD68 阳性巨噬细胞在肾组织中的浸润。