Department of Critical care medicine, Chengdu Second People's Hospital; Sichuan, China.
Department of Critical care medicine, The second Affiliated Hospital of Chongqing Medical University; Chongqing, China.
J Surg Res. 2021 Sep;265:223-232. doi: 10.1016/j.jss.2021.03.047. Epub 2021 May 3.
Long non-coding RNAs (lncRNAs) have been demonstrated to be involved in the progression of sepsis-induced acute kidney injury (AKI). In this study, we aimed to explore the functions of lncRNA cancer susceptibility candidate 2 (CASC2) in sepsis-induced AKI.
The sepsis cell models were established by exposing HK2 and HEK293 cells into lipopolysaccharide (LPS). Quantitative real-time polymerase chain reaction (qRT-PCR) assay was conducted to determine the expression of CASC2, miR-545-3p and peroxisome proliferator-activated receptor-α (PPARA) mRNA. Cell Counting Kit-8 (CCK-8) assay, flow cytometry analysis and wound healing assay were employed for cell viability, apoptosis and migration, respectively. Western blot assay was conducted for the protein levels of E-cadherin, α-SMA and PPARA. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by specific kits. The relationship between miR-545-3p and CASC2 or PPARA was verified by dual-luciferase reporter assay.
CASC2 level was decreased in sepsis patients' serums and LPS-treated HK2 and HEK293 cells. CASC2 overexpression facilitated cell viability and restrained cell apoptosis, migration, epithelial-mesenchymal transition (EMT) and oxidative stress in LPS-triggered HK2 and HEK293 cells. CASC2 was identified as a sponge for miR-545-3p to regulate PPARA expression. MiR-545-3p overexpression restored the impact of CASC2 on LPS-induced injury in HK2 and HEK293 cells. Moreover, miR-545-3p overexpression aggravated LPS-induced cell injury in HK2 and HEK293 cells by targeting PPARA.
CASC2 overexpression relieved the damage of HK2 and HEK293 cells mediated by LPS treatment through regulating miR-545-3p/PPARA axis.
长链非编码 RNA（lncRNA）已被证明参与了脓毒症诱导的急性肾损伤（AKI）的进展。在这项研究中，我们旨在探讨 lncRNA 癌症易感性候选物 2（CASC2）在脓毒症诱导的 AKI 中的作用。
通过将 HK2 和 HEK293 细胞暴露于脂多糖（LPS）来建立脓毒症细胞模型。通过定量实时聚合酶链反应（qRT-PCR）测定 CASC2、miR-545-3p 和过氧化物酶体增殖物激活受体-α（PPARA）mRNA 的表达。通过细胞计数试剂盒-8（CCK-8）测定、流式细胞术分析和划痕愈合试验分别测定细胞活力、凋亡和迁移。通过 Western blot 测定 E-钙黏蛋白、α-SMA 和 PPARA 的蛋白水平。通过特定试剂盒测定超氧化物歧化酶（SOD）和丙二醛（MDA）的水平。通过双荧光素酶报告基因实验验证 miR-545-3p 与 CASC2 或 PPARA 之间的关系。
CASC2 水平在脓毒症患者的血清和 LPS 处理的 HK2 和 HEK293 细胞中降低。CASC2 过表达促进了 LPS 触发的 HK2 和 HEK293 细胞中的细胞活力，并抑制了细胞凋亡、迁移、上皮-间充质转化（EMT）和氧化应激。CASC2 被鉴定为 miR-545-3p 的海绵体，以调节 PPARA 表达。miR-545-3p 过表达恢复了 CASC2 对 LPS 诱导的 HK2 和 HEK293 细胞损伤的影响。此外，miR-545-3p 过表达通过靶向 PPARA 加重了 LPS 诱导的 HK2 和 HEK293 细胞损伤。
CASC2 过表达通过调节 miR-545-3p/PPARA 轴缓解了 LPS 处理介导的 HK2 和 HEK293 细胞的损伤。
