青蒿琥酯通过Keap1/Nrf2通路抑制氧化应激改善犬急性肾损伤的体内外分析

doi:10.11843/j.issn.0366-6964.2022.07.031

Abstract

Abstract: The aim of this study was to investigate the antioxidant effect of artesunate (ART) on acute kidney injury induced by gentamicin (GM) in dogs and its mechanism. Twenty dogs were randomly divided into 4 groups:control group (Control), gentamicin model group (GM), artesunate group (GM+ART), artesunate + ML385 group (GM+ART + ML385). In addition to the control group, other groups were injected with GM to establish AKI model. After successful modeling, Dogs in GM+ART group were given ART, and GM+ART+ML385 group were given ART and ML385 at the same time. The control group and GM group were given the same volume of normal saline. The test period is 12 days. Different concentrations of GM were co-cultured with MDCK cells, and the optimal concentration was 4.0 mmol·L^-1. The optimum concentration of ART was determined to be 50.0 μ mol·L^-1 by the same method. The MDCK cells, MDCK cells overexpressing kelch like ECH related protein 1 (Keap1) (M-K) and MDCK cells knockdown nuclear factor E2 related factor 2 (Nrf2) (M-SiNrf2) were divided into three groups respectively:Healthy control group, GM control group and GM + ART intervention group, co-culturing for 24 hours. The results show as follows:1) In animal experiments, the levels of Cr, UN and KIM-1 in GM + ART group were significantly lower than those in GM Group, and the levels of Cr, UN and KIM-1 in GM + ART + ML385 group were significantly higher than those in GM + ART group. 2) In animal experiments, compared with GM Group, the expression of Nrf2 and glutathione cysteine ligase catalytic subunit (GCLC) protein was up-regulated, Keap1 protein was down-regulated, the levels of total superoxide dismutase (T-SOD) and glutathione (GSH) in renal homogenate were increased, and the level of malondialdehyde (MDA) was decreased in GM + ARTgroup. Compared with GM + ART group, the expression of Nrf2 and GCLC decreased, and the levels of T-SOD and GSH decreased after intervening by ML385. 3) In the cell test, compared with MDCK cells co cultured with 4.0 mmol·L^-1 GM, the dose of 50.0 μmol·L^-1 ART added could significantly increase the cell proliferation rate, reduce the level of ROS, down regulate the expression of Keap1 and up regulate the expression of Nrf2, heme oxygenase 1 (HO1) and GCLC. 4) In the cell test, compared with MDCK cells, the expression of Keap1 was up-regulated, the expression of Nrf2, HO1 and GCLC were down regulated, the cell proliferation rate was decreased, and the content of ROS was increased both in M-K cells and M-SiNrf2 cells under the same treatment of GM + ART(P<0.05 or P<0.01). In conclusion, artesunate has a protective effect on acute renal injury induced by gentamicin in dogs, and its mechanism is related to inhibit oxidative stress via Keap1/Nrf2 signaling pathway.

Key words: artesunate, gentamicin, dog, Keap1/Nrf2 signaling pathway, oxidative stress, acute kidney injury

CLC Number:

S856.5

LU Jiang, ZHU Daoxian, LIU Li, HAO Fuxing, WU Zhi, FU Hongqing. Artesunate Improves Acute Renal Injury in Dogs by Inhibiting Oxidative Stress Via Keap1/Nrf2 Signaling Pathway in vitro and in vivo[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2343-2353.

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Artesunate Improves Acute Renal Injury in Dogs by Inhibiting Oxidative Stress Via Keap1/Nrf2 Signaling Pathway in vitro and in vivo

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