Department of Anesthesiology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Department of Hand Surgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
J Gene Med. 2021 Aug;23(8):e3349. doi: 10.1002/jgm.3349. Epub 2021 Jun 25.
Propofol (PPF) plays a protective role in ischemia-reperfusion (I/R) in multiple organs, including renal ischemia-reperfusion injury (RIRI). The present study aimed to investigate the underlying mechanisms by which PPF exerts its protective functions in RIRI.
BALB/c mice were employed for the construction of RIRI animal model. PPF pre-treatment was carried out before I/R. An in vitro I/R model was established with HK-2 cells after hypoxia/reoxygenation (H/R) culture, and PPF was utilized to treat the cells before H/R. A quantitative-polymerase chain reaction (qPCR) was conducted to detect long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and miR-126-5p expression levels. Flow cytometry was adopted to detect the apoptosis of HK-2 cells. Bioinformatics analysis, qPCR, a luciferase reporter gene experiment and a RNA immunoprecipitation experiment were used to determine the regulatory relationship between MALAT1 and miR-126-5p. The expression level of vascular endothelial growth factor A (VEGFA) was examined by western blotting.
MALAT1 expression was augmented and miR-126-5p was decreased in RIRI models. PPF pre-treatment remarkably reduced creatinine and urea nitrogen levels in the serum of BALB/c mice with RIRI, and diminished the apoptosis of HK-2 cells treated with H/R. In addition, PPF pre-treatment markedly restrained the expression of MALAT1 in both in vivo and in vitro models and up-regulated miR-126-5p expression. MALAT1 could adsorb miR-126-5p to repress it and up-regulate VEGFA. MALAT1 overexpression reversed the protective effects of PPF on RIRI.
PPF protects the kidney against RIRI by inhibiting MALAT1 and up-regulating miR-126-5p expression, as well as indirectly inhibiting the expression of VEGFA.
丙泊酚（PPF）在包括肾缺血再灌注损伤（RIRI）在内的多种器官的缺血再灌注（I/R）中发挥保护作用。本研究旨在探讨 PPF 发挥其在 RIRI 中保护作用的潜在机制。
使用 BALB/c 小鼠构建 RIRI 动物模型。在 I/R 前进行 PPF 预处理。用缺氧/复氧（H/R）培养后的 HK-2 细胞建立体外 I/R 模型，并在 H/R 前用 PPF 处理细胞。采用定量聚合酶链反应（qPCR）检测长链非编码 RNA 转移相关肺腺癌转录物 1（MALAT1）和 miR-126-5p 的表达水平。采用流式细胞术检测 HK-2 细胞的凋亡情况。通过生物信息学分析、qPCR、荧光素酶报告基因实验和 RNA 免疫沉淀实验确定 MALAT1 和 miR-126-5p 之间的调控关系。采用 Western blot 检测血管内皮生长因子 A（VEGFA）的表达水平。
在 RIRI 模型中，MALAT1 的表达增加，miR-126-5p 的表达减少。PPF 预处理可显著降低 RIRI 模型 BALB/c 小鼠血清中肌酐和尿素氮水平，并减少 H/R 处理的 HK-2 细胞凋亡。此外，PPF 预处理可显著抑制体内和体外模型中 MALAT1 的表达并上调 miR-126-5p 的表达。MALAT1 可以吸附 miR-126-5p 来抑制它并上调 VEGFA。MALAT1 的过表达逆转了 PPF 对 RIRI 的保护作用。
PPF 通过抑制 MALAT1 和上调 miR-126-5p 的表达，以及间接抑制 VEGFA 的表达，来保护肾脏免受 RIRI 损伤。
