Department of Emergency Intensive Care Unit, Zhejiang Province People's Hospital, Hangzhou, Zhejiang, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5592-5603. doi: 10.26355/eurrev_202005_21345.
Lipopolysaccharide (LPS)-induced inflammation and dysfunction in the kidney may be the major risk factors for subsequent acute kidney injury (AKI). Previous studies have reported that up-regulation of notch receptor 3 (NOTCH3) expression is accompanied with renal epithelium and podocyte damage. Herein, we aimed to investigate whether NOTCH3 was involved in lipopolysaccharide (LPS)-induced AKI and renal cell dysfunction.
Septic mice were established using LPS (20 mg/kg) intraperitoneally. mRNA and protein expression in the kidney and renal cell was performed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting, respectively. Cell counting kit-8 (CCK8) and flow cytometry were used to measure cell viability and apoptosis, respectively. Bioinformatics algorithm and Luciferase reporter gene assay were performed to validate whether NOTCH3 was a direct target of miR-201-5p.
Up-regulation of NOTCH3 and down-regulation of miR-201-5p were observed in the kidney of LPS-induced septic mice. LPS-stimulated TCMK-1 and MPC5 cells led to an increase in NOTCH3 and a decrease in miR-201-5p expression levels. Bioinformatics algorithm and experimental measurements validated that NOTCH3 was a direct target of miR-201-5p. Overexpression of miR-201-5p protected against LPS-induced renal cell growth inhibition, apoptosis and inflammatory response via the suppression of toll-like receptor 4 (TLR4)/NOTCH3 signaling pathway.
The novel role of miR-201-5p via the inhibition of LPS-activated TLR4/NOTCH3 might provide a potential therapeutic strategy for the treatment of LPS-induced AKI.
脂多糖（LPS）诱导的肾脏炎症和功能障碍可能是随后发生急性肾损伤（AKI）的主要危险因素。先前的研究报告称，NOTCH3 受体表达上调伴随着肾上皮细胞和足细胞损伤。在此，我们旨在研究 NOTCH3 是否参与 LPS 诱导的 AKI 和肾细胞功能障碍。
通过 LPS（20mg/kg）腹腔内注射建立脓毒症小鼠模型。采用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot 分别检测肾脏和肾细胞中的 mRNA 和蛋白表达。细胞计数试剂盒-8（CCK8）和流式细胞术分别用于测量细胞活力和细胞凋亡。生物信息学算法和荧光素酶报告基因检测用于验证 NOTCH3 是否是 miR-201-5p 的直接靶标。
在 LPS 诱导的脓毒症小鼠肾脏中观察到 NOTCH3 的上调和 miR-201-5p 的下调。LPS 刺激 TCMK-1 和 MPC5 细胞导致 NOTCH3 增加和 miR-201-5p 表达水平降低。生物信息学算法和实验测量验证了 NOTCH3 是 miR-201-5p 的直接靶标。miR-201-5p 的过表达通过抑制 Toll 样受体 4（TLR4）/NOTCH3 信号通路，可防止 LPS 诱导的肾细胞生长抑制、凋亡和炎症反应。
miR-201-5p 通过抑制 LPS 激活的 TLR4/NOTCH3 的新作用可能为治疗 LPS 诱导的 AKI 提供一种潜在的治疗策略。
