miR-34b-5p 通过抑制水通道蛋白-2 促进脓毒症诱导的急性肾损伤中的肾细胞炎症和凋亡。
miR-34b-5p promotes renal cell inflammation and apoptosis by inhibiting aquaporin-2 in sepsis-induced acute kidney injury.
机构信息
Department of Critical Care Medicine, Fujian Provincial Hospital, Fuzhou, P.R. China.
School of Clinical Medicine, Fujian Medical University, Fuzhou, P.R. China.
出版信息
Ren Fail. 2021 Dec;43(1):291-301. doi: 10.1080/0886022X.2021.1871922.
OBJECTIVE
This study was designed to uncover the mechanism of miR-34b-5p-mediated aquaporin-2 (AQP2) in sepsis-induced injury using human renal tubular epithelial cells (HK-2).
METHODS
Serum levels of miR-34b-5p, TNF-α, IL-1β, IL-6, serum creatinine (SCr), and blood urea nitrogen (BUN) in septic patients with acute kidney injury (AKI) and healthy controls were detected. Lipopolysaccharide (LPS) was used to induce sepsis in HK-2 cells. LPS-induced HK-2 cells were transfected with miR-34b-5p inhibitor, miR-34b-5p mimic, pcDNA3.1-AQP2, si-AQP2, miR-34b-5p inhibitor + si-NC, or miR-34b-5p inhibitor + si-AQP2. The expressions of miR-34b-5p, AQP2, Bax, Bcl-2, cleaved caspase-3, TNF-α, IL-1β, and IL-6 in HK-2 cells were detected. TUNEL staining revealed the apoptosis of HK-2 cells. Dual-luciferase reporter assay verified the binding between miR-34b-5p and AQP2.
RESULTS
The expression of miR-34b-5p and the inflammatory responses were augmented in septic AKI patients. miR-34b-5p was up-regulated and AQP2 was down-regulated in LPS-induced HK-2 cells. miR-34b-5p inhibition or AQP2 overexpression ameliorated apoptosis and inflammation in LPS-induced HK-2 cells. In contrast, overexpressing miR-34b-5p deteriorated LPS-induced injury in HK-2 cells. AQP2 was a downstream target of miR-34b-5p. AQP2 silencing abolished the suppressive effects of miR-34b-5p inhibition on LPS-induced apoptosis and inflammatory response in HK-2 cells.
CONCLUSION
miR-34b-5p inhibits AQP2 to promote LPS-induced injury in HK-2 cells.
目的
本研究旨在利用人肾小管上皮细胞（HK-2）揭示 miR-34b-5p 介导水通道蛋白-2（AQP2）在脓毒症诱导损伤中的作用机制。
方法
检测脓毒症合并急性肾损伤（AKI）患者和健康对照者血清中 miR-34b-5p、TNF-α、IL-1β、IL-6、血清肌酐（SCr）和血尿素氮（BUN）水平。用脂多糖（LPS）诱导 HK-2 细胞发生脓毒症。将 miR-34b-5p 抑制剂、miR-34b-5p 模拟物、pcDNA3.1-AQP2、si-AQP2、miR-34b-5p 抑制剂+si-NC 或 miR-34b-5p 抑制剂+si-AQP2 转染 LPS 诱导的 HK-2 细胞。检测 HK-2 细胞中 miR-34b-5p、AQP2、Bax、Bcl-2、cleaved caspase-3、TNF-α、IL-1β 和 IL-6 的表达。TUNEL 染色显示 HK-2 细胞的凋亡情况。双荧光素酶报告基因实验验证 miR-34b-5p 与 AQP2 之间的结合。
结果
脓毒症 AKI 患者中 miR-34b-5p 的表达和炎症反应增强。LPS 诱导的 HK-2 细胞中 miR-34b-5p 上调，AQP2 下调。miR-34b-5p 抑制或 AQP2 过表达可改善 LPS 诱导的 HK-2 细胞凋亡和炎症。相反，过表达 miR-34b-5p 加重 LPS 诱导的 HK-2 细胞损伤。AQP2 是 miR-34b-5p 的下游靶标。AQP2 沉默消除了 miR-34b-5p 抑制对 LPS 诱导的 HK-2 细胞凋亡和炎症反应的抑制作用。
结论
miR-34b-5p 通过抑制 AQP2 促进 LPS 诱导的 HK-2 细胞损伤。
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