ABSTRACT

This study explored the mechanism of NEAT1 in sepsis-induced AKI rats. Cecal ligation punctures (CLP)-induced AKI rats were injected with siRNA-NEAT1 lentivirus. Kidney histopathology and apoptosis were evaluated via hematoxylin-eosin and TUNEL staining, respectively. ELISA determined the levels of Blood urea nitrogen (BUN), serum creatinine (SCr), neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), TNF-α, Interleukin (IL)-1β, and IL-6. Colorimetry measured malondialdehyde (MDA), superoxide dismutase (SOD) activities. qPCR analyzed NEAT1, miR-27a-3p, TAB3, Bcl-2, and Bax expressions. siNEAT1 reversed the promotive effect of CLP on kidney histopathological injury, and BUN, SCr, NGAL, KIM-1, TNF-α, IL-1β, IL-6, MDA, and Bax levels and apoptosis, but raised CLP-downregulated SOD and Bcl-2 levels. NEAT1 sponged miR-27a-3p which targeted TAB3. siNEAT1 upregulated miR-27a-3p and downregulated TAB3 expression. TAB3 overexpression reversed the inhibitory effect of siNEAT1 on the LPS-induced apoptosis of HK-2 cells. siNEAT1 alleviated sepsis-induced AKI in rats and LPS-induced sepsis of cells via miR-27a-3p/TAB3 axis.

摘要

本研究探讨了 NEAT1 在脓毒症诱导的 AKI 大鼠中的作用机制。盲肠结扎穿刺 (CLP) 诱导的 AKI 大鼠被注射 siRNA-NEAT1 慢病毒。肾组织病理学和细胞凋亡分别通过苏木精-伊红和 TUNEL 染色进行评估。ELISA 测定血尿素氮 (BUN)、血清肌酐 (SCr)、中性粒细胞明胶酶相关载脂蛋白 (NGAL)、肾损伤分子-1 (KIM-1)、TNF-α、白细胞介素 (IL)-1β 和IL-6。比色法测量丙二醛 (MDA)、超氧化物歧化酶 (SOD) 的活性。qPCR 分析了 NEAT1、miR-27a-3p、TAB3、Bcl-2 和 Bax 的表达。siNEAT1 逆转 CLP 对肾脏组织病理学损伤的促进作用，以及 BUN、SCr、NGAL、KIM-1、TNF-α、IL-1β、IL-6、MDA 和 Bax 水平和细胞凋亡，但提高 CLP 下调的 SOD 和Bcl-2 水平。NEAT1 吸收靶向 TAB3 的 miR-27a-3p。siNEAT1 上调 miR-27a-3p 并下调 TAB3 表达。TAB3 过表达逆转了 siNEAT1 对 LPS 诱导的 HK-2 细胞凋亡的抑制作用。siNEAT1 通过 miR-27a-3p/TAB3 轴减轻大鼠脓毒症诱导的 AKI 和 LPS 诱导的细胞脓毒症。