DataSheet1_Analysis of exosome-derived microRNAs as early biomarkers of lipopolysaccharide-induced acute kidney injury in rats.docx

Sepsis contributes to the high prevalence of acute kidney injury (AKI), which mainly occurs in hospitalized patients. The delay in AKI detection is a risk factor for death and chronicity; thus, early diagnosis is essential for initiating proper treatment strategies. Although serum creatinine is used as biomarker, it is increased in plasma serum creatinine only at late stages of AKI. MicroRNAs (miRNAs), a class of noncoding RNAs responsible for gene regulation, can be found in biological fluids within vesicles such as exosomes and may be promising tools for the early detection of AKI. We aimed to identify potential blood miRNAs that can be used as early biomarkers of sepsis-induced AKI in rats. Adult male Wistar rats received a single dose of lipopolysaccharide (LPS). The earliest significant increase in serum creatinine was detected 4 h after LPS administration. To evaluate whether miRNAs could act as early biomarkers, blood samples were collected before and 2 h after LPS infusion. Serum NGAL levels were used as a comparative marker. Serum miRNAs were derived from exosomes, and their expression were evaluated by the PCR array. miR-181a-5p and miR-23b-3p showed higher expression in LPS-treated rats than in the control animals (p < 0.05). Bioinformatics analysis showed that both miRNAs target molecules associated with transcription factors that regulate genes related to proinflammatory cytokines. Considering that LPS activates transcription factors that lead to the production of proinflammatory cytokines, possible premature changes in the serum levels of miR-181a-5p and miR-23b-3p may be used to identify sepsis-induced AKI earlier.

脓毒症（Sepsis）是急性肾损伤（acute kidney injury, AKI）高患病率的重要诱因，该病主要累及住院患者。AKI检测延迟是导致患者死亡及病情慢性化的风险因素，因此早期诊断对于启动合理治疗方案至关重要。尽管目前临床以血清肌酐作为生物标志物，但该指标仅在AKI晚期阶段才会出现水平升高。微小RNA（MicroRNAs, miRNAs）是一类参与基因调控的非编码RNA，可存在于外泌体等囊泡包裹的生物体液中，有望成为AKI早期检测的潜在工具。本研究旨在筛选可作为大鼠脓毒症诱导型AKI早期生物标志物的潜在血液miRNAs。成年雄性Wistar大鼠单次注射脂多糖（lipopolysaccharide, LPS）。在LPS给药后4小时，首次检测到血清肌酐出现显著升高。为评估miRNAs是否可作为早期生物标志物，我们在LPS输注前及输注后2小时采集血液样本，并以血清中性粒细胞明胶酶相关脂质运载蛋白（Neutrophil Gelatinase-Associated Lipocalin, NGAL）水平作为对照标志物。血清miRNAs提取自外泌体，通过PCR芯片（PCR array）检测其表达水平。与对照组大鼠相比，LPS处理组大鼠的miR-181a-5p与miR-23b-3p表达水平显著升高（p < 0.05）。生物信息学分析表明，这两种miRNAs的靶分子均与调控促炎细胞因子相关基因的转录因子有关。鉴于LPS可激活介导促炎细胞因子产生的转录因子，因此血清miR-181a-5p与miR-23b-3p水平的早期变化，或可用于更早识别脓毒症诱导型AKI。