Trama Center/Department of Emergency and Trauma Surgery, Tongji Hospital Tongji Medical College, Huazhong University of Science and Technology, China.
Department of Oncology, Taikang Tongji (Wuhan) Hospital, China.
Int Immunopharmacol. 2020 Nov;88:106965. doi: 10.1016/j.intimp.2020.106965. Epub 2020 Sep 18.
We aimed to study the effects and the underlying mechanisms of Diosmetin (DIOS) in rats with sepsis-induced acute kidney injury (AKI).
The AKI model in RMCs was induced using LPS, and the cells were then treated with DIOS. Cell viability, apoptosis, inflammatory response, and antioxidant were measured using MTT, Flow cytometry, ELISA, and Lucigenin assay, respectively. The correlation between TUG1 and Nrf2 was confirmed by RNA pull-down and RNA immunoprecipitation. Real-time quantitative PCR and Western blot were performed to detect the expressions of gene and proteins during the development of AKI. The effects of lncRNA-TUG1 silencing and Nrf2 silencing on cell physiological functions were detected. Moreover, a rat sepsis-induced AKI model followed by Hematoxylin & Eosin (H&E) and immunofluorescence staining were performed.
The experimental concentration of DIOS was determined to be 20 μM. After LPS treatment, the activity of RMCs was decreased, the apoptosis rate, inflammation and oxidative stress damage were increased, moreover, the expression of Nrf2/HO-1 signal axis was inhibited and caspase-3 was activated. However, DIOS significantly reversed these effects caused by LPS treatment, and increased the expression of lncRNA-TUG1, but lncRNA-TUG1 silencing effectively reversed the effects of DIOS. In addition, lncRNA-TUG1 was found to interact with Nrf2. Overexpression of TUG1 could reduce the damage of LPS caused to cell physiological functions, which were reversed by siNrf2. Thus, DIOS treatment could improve the physiological and pathological damages of renal tissues in AKI rats.
DIOS may reduce sepsis-induced AKI through enhancing the TUG1/Nrf2/HO-1 pathway.
本研究旨在探讨地奥司明（DIOS）对脂多糖诱导的急性肾损伤（AKI）大鼠的作用及其机制。
采用脂多糖（LPS）诱导 RMCs 发生 AKI 模型，用 DIOS 处理细胞。MTT、流式细胞术、ELISA 和 Lucigenin 测定法分别用于检测细胞活力、凋亡、炎症反应和抗氧化能力。采用 RNA 下拉和 RNA 免疫沉淀法证实 TUG1 与 Nrf2 之间的相关性。实时定量 PCR 和 Western blot 用于检测 AKI 发展过程中基因和蛋白的表达。检测沉默 lncRNA-TUG1 和 Nrf2 对细胞生理功能的影响。此外，还进行了大鼠脓毒症诱导的 AKI 模型，随后进行苏木精和伊红（H&E）和免疫荧光染色。
确定 DIOS 的实验浓度为 20 μM。LPS 处理后，RMC 活性降低，凋亡率、炎症和氧化应激损伤增加，Nrf2/HO-1 信号轴表达受抑制，caspase-3 被激活。然而，DIOS 能显著逆转 LPS 处理引起的这些作用，并增加 lncRNA-TUG1 的表达，但 lncRNA-TUG1 沉默能有效逆转 DIOS 的作用。此外，发现 lncRNA-TUG1 与 Nrf2 相互作用。TUG1 的过表达可减轻 LPS 对细胞生理功能的损伤，而 siNrf2 则可逆转这一损伤。因此，DIOS 治疗可改善 AKI 大鼠肾组织的生理和病理损伤。
DIOS 可能通过增强 TUG1/Nrf2/HO-1 通路减轻脓毒症引起的 AKI。
