长链非编码 RNA-MIAT 通过调控 miR-29a 表达对脓毒症大鼠肾损伤的影响。
Effect of lncRNA-MIAT on kidney injury in sepsis rats via regulating miR-29a expression.
机构信息
Department of Critical Care Medicine, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Dec;23(24):10942-10949. doi: 10.26355/eurrev_201912_19797.
OBJECTIVE
The long non-coding ribonucleic acid (lncRNA)-myocardial infarction associated transcript (MIAT) has been demonstrated to serve as a key regulator in various physiological and pathological processes. This study aims to explore whether lncRNA-MIAT regulates the expression of micro RNA (miR)-29a to affect kidney's injury in sepsis rats.
MATERIALS AND METHODS
A total of 30 healthy male Sprague-Dawley (SD) rats were randomly divided into experimental group and control group. Rats in the experimental group were injected with lipopolysaccharide (LPS) through the tail vein to prepare the model of sepsis-induced kidney injury, while those in the control group with the equal volume of normal saline. After the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) in rats were determined to ascertain successful modeling, fluorescence quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to measure the expression levels of the lncRNA-MIAT and miR-29a messenger RNAs (mRNAs) in renal tissues. The normal rat kidney epithelial (NRK-52E) cell line was cultured in vitro, and the model was established in vitro via LPS to study the influences of lncRNA-MIAT and miR-29a on the kidney injury in sepsis rats. Moreover, cell apoptosis was detected using Western blotting.
RESULTS
According to the results of the rat in vivo experiment and NRK-52E cell line in vitro experiment, the model of kidney injury was established successfully, and compared with the control group, experiment group had significantly raised SCr and BUN levels (p<0.01) and a remarkably increased lincRNA-MIAT gene expression level (p<0.01), but a substantially decreased miR-29a gene expression level (p<0.01). Additionally, when the expression of lncRNA-MIAT was up-regulated, the expression of miR-29a was prominently decreased (p<0.01), but that of the cell apoptosis gene cysteine-aspartic proteases (Caspase)-8 protein was remarkably increased (p<0.01). However, the expression of Caspase-8 protein was significantly lowered (p<0.01) once the expression of miR-29a was up-regulated.
CONCLUSIONS
LncRNA-MIAT may bind to miR-29a to participate in sepsis-related kidney injury.
目的
长链非编码核糖核酸（lncRNA）-心肌梗塞相关转录物（MIAT）已被证明在各种生理和病理过程中作为关键调节剂发挥作用。本研究旨在探讨 lncRNA-MIAT 是否通过调节 microRNA（miR）-29a 的表达来影响脓毒症大鼠的肾脏损伤。
材料和方法
将 30 只健康雄性 Sprague-Dawley（SD）大鼠随机分为实验组和对照组。实验组大鼠通过尾静脉注射脂多糖（LPS）制备脓毒症诱导的肾损伤模型，而对照组大鼠注射等体积的生理盐水。大鼠血清肌酐（SCr）和血尿素氮（BUN）水平确定模型成功建立后，采用荧光定量实时聚合酶链反应（qRT-PCR）测定肾组织中 lncRNA-MIAT 和 miR-29a 信使 RNA（mRNA）的表达水平。体外培养正常大鼠肾上皮（NRK-52E）细胞系，通过 LPS 建立体外模型，研究 lncRNA-MIAT 和 miR-29a 对脓毒症大鼠肾损伤的影响。此外，还通过 Western blot 检测细胞凋亡。
结果
根据大鼠体内实验和 NRK-52E 细胞系体外实验结果，成功建立了肾损伤模型，与对照组相比，实验组大鼠的 SCr 和 BUN 水平明显升高（p<0.01），lncRNA-MIAT 基因表达水平明显升高（p<0.01），miR-29a 基因表达水平明显降低（p<0.01）。此外，上调 lncRNA-MIAT 的表达时，miR-29a 的表达明显降低（p<0.01），胱天蛋白酶-8 蛋白（Caspase）-8 基因的表达明显升高（p<0.01）。然而，上调 miR-29a 的表达后，Caspase-8 蛋白的表达明显降低（p<0.01）。
结论
lncRNA-MIAT 可能通过与 miR-29a 结合参与脓毒症相关的肾损伤。
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