Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Engineering Research Center of Natural Polymer-based Medical Materials in Hubei Province, Wuhan, China.
Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Engineering Research Center of Natural Polymer-based Medical Materials in Hubei Province, Wuhan, China.
Life Sci. 2020 Jul 15;253:117705. doi: 10.1016/j.lfs.2020.117705. Epub 2020 Apr 22.
Ischemia-reperfusion injury (IRI) is harmful to patients following kidney transplantation. Hypothermic machine perfusion (HMP) can be adopted to preserve grafts and reduce consequential injury. We hypothesized that aldehyde dehydrogenase 2 (ALDH2) partly mitigates kidney IRI via regulating excessive autophagy in HMP.
The rabbits were assigned to 5 groups: Normal, HMP, HMP + Alda-1, HMP + CYA and cold storage (CS). After the rabbit autologous kidney transplantation, renal pathology and function were evaluated by histological analysis, glomerular related proteins (desmin, nephrin), tubular injury factors (NGAL, Ki67), serum creatinine (Cr) and blood urea nitrogen (BUN). Oxidative stress molecular Malondialdehyde (MDA) and superoxide dismutase (SOD2) expression, as well as inflammatory cytokines (TNF-α, IL-6, IL-10) were assessed by immunohistochemistry. The expression of LC3, p62, ALDH2, p-Akt, mTOR, PTEN, p-PTEN, and 4-HNE were measured by immunohistochemistry, RT-PCR, Western blot analysis or ELISA.
HMP was more effective than CS for kidney preservation, with p- ALDH2 expressed in greater quantities in HMP. The results of kidney pathology and function in HMP + Alda-1 were the best. The MDA & SOD2 and the Vyacheslav score were improved in HMP + CYA. ALDH2 reduced 4-HNE-induced oxidative stress, inflammatory infiltration, the expression of LC3, p62 and inhibited autophagy accompanied by activation of p-Akt and mTOR via p-PTEN/PTEN.
Akt-mTOR autophagy pathway is a novel target for ALDH2 to reduce renal IRI partly by inhibition of 4-HNE in HMP, then protecting the donated kidney received after cardiac death (DCD).
缺血再灌注损伤（IRI）对肾移植后的患者有害。低温机器灌注（HMP）可用于保存移植物并减少后续损伤。我们假设醛脱氢酶 2（ALDH2）通过调节 HMP 中过度的自噬来部分减轻肾 IRI。
兔子被分为 5 组：正常组、HMP 组、HMP+Alda-1 组、HMP+CYA 组和冷藏（CS）组。在兔自体肾移植后，通过组织学分析、肾小球相关蛋白（波形蛋白、nephrin）、肾小管损伤因子（NGAL、Ki67）、血清肌酐（Cr）和血尿素氮（BUN）评估肾脏病理和功能。通过免疫组织化学评估氧化应激分子丙二醛（MDA）和超氧化物歧化酶 2（SOD2）的表达，以及炎症细胞因子（TNF-α、IL-6、IL-10）。通过免疫组织化学、RT-PCR、Western blot 分析或 ELISA 测量 LC3、p62、ALDH2、p-Akt、mTOR、PTEN、p-PTEN 和 4-HNE 的表达。
HMP 比 CS 更有效地保存肾脏，HMP 中表达更多的 p-ALDH2。HMP+Alda-1 的肾脏病理和功能结果最好。HMP+CYA 改善了 MDA 和 SOD2 以及 Vyacheslav 评分。ALDH2 通过 p-PTEN/PTEN 减少 4-HNE 诱导的氧化应激、炎症浸润、LC3、p62 的表达，并抑制自噬，同时激活 p-Akt 和 mTOR。
Akt-mTOR 自噬途径是 ALDH2 的一个新靶点，通过抑制 HMP 中的 4-HNE，部分减轻心脏死亡（DCD）后接受的供肾的肾 IRI。
