安徽医科大学学报2024，Vol.59Issue(12)：2079-2086,8.DOI:10.19405/j.cnki.issn1000-1492.2024.12.003
MIF抑制剂ISO-1通过减轻氧化应激、炎症反应和细胞凋亡缓解脓毒症诱导的急性肾损伤
MIF inhibitor ISO-1 alleviates sepsis-induced acute kidney injury by suppressing oxidative stress,inflammation,and apoptosis
摘要
Abstract
Objective To investigate the effects and underlying mechanisms of the macrophage migration inhibitory factor(MIF)inhibitor(S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazoleacetic acid methyl ester(ISO-1)on sepsis-induced acute kidney injury(AKI).Methods Human renal tubular epithelial HK-2 cells were divided into Con group(without any treatment),ISO-1 group(10 μg/ml ISO-1 treatment for 24 h)and LPS group(10 µg/ml LPS treatment for 24 h),LPS+ISO-1 group(10 μg/ml LPS treatment for 24 h followed by 10 μg/ml ISO-1 treat-ment for 24 h).ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in the cell supernatants.Reactive oxygen species(ROS)levels were assessed u-sing the 6-carboxyl-2',7'-dichlorodihydrofluorescein diacetate fluorescent indicator(DCFH-DA)method.Apopto-sis levels were detected by TUNEL staining,and Western blot was employed to analyze the expression of proteins of Kelch like ECH associated protein 1(Keap1),NFE2 like bZIP transcription factor 2(Nrf2),heme oxygenase-1(HO-1),as well as apoptosis-related proteins Bcl-2,Bax,and cleaved Caspase-3(c-Caspase-3).A sepsis mouse model was established using the cecal ligation and puncture(CLP)method,and the mice were divided into four groups:sham-operated(Sham),ISO-1 control(ISO-1),CLP,and ISO-1 treatment(CLP+ISO-1).After the ex-periment,mouse kidney tissues were collected for HE staining to observe pathological changes.Blood urea nitrogen(BUN),serum creatinine(Scr),myeloperoxidase(MPO)levels in kidney tissues,glutathione(GSH)and su-peroxide dismutase(SOD)activities were measured.Western blot was also used to detect the expression of MIF and proteins in the Nrf2/Keap1 signaling pathway and apoptosis-related proteins in kidney tissues.Results Com-pared to the Con group,the LPS and LPS+ISO-1 groups showed significantly increased levels of TNF-α,IL-1β,IL-6,TUNEL-positive rates,ROS levels,and protein expressions of Keap1,Bax,and c-Caspase-3 in HK-2 cells(P＜0.05),while the expressions of Nrf2,HO-1,and Bcl-2 significantly decreased(P＜0.05).The ISO-1 group showed no significant changes(P＞0.05).Compared to the LPS group,the LPS+ISO-1 group exhibited significantly decreased levels of TNF-α,IL-1β,IL-6,TUNEL-positive rates,ROS levels,and protein expressions of Keap1,Bax,and c-Caspase-3,while the expressions of Nrf2,HO-1,and Bcl-2 significantly increased(P＜0.05).In the mouse experiments,compared to the Sham group,the CLP and CLP+ISO-1 groups showed severe kidney tissue damage,increased levels of serum BUN,Scr,and kidney MIF,Keap1,Bax,and c-Caspase-3 pro-tein expressions(P＜0.05),while GSH,SOD activities,and protein expressions of Nrf2,HO-1,and Bcl-2 sig-nificantly decreased(P＜0.05).The ISO-1 group showed no significant changes(P＞0.05).Compared to the CLP group,the CLP+ISO-1 group showed significant improvements in the aforementioned indicators(P＜0.05).Conclusion The specific MIF inhibitor ISO-1 can ameliorate sepsis-induced AKI by inhibiting oxidative stress,in-flammatory response,and apoptosis both in vitro and in vivo.The mechanism may be through Nrf2/Keap1 signaling pathway.关键词
脓毒症/急性肾损伤/巨噬细胞移动抑制因子拮抗剂ISO-1/氧化应激/炎症/细胞凋亡Key words
sepsis/acute kidney injury/macrophage migration inhibitor antagonist ISO-1/oxidative stress/in-flammation/apoptosis分类
医药卫生引用本文复制引用
殷富康,张晓霞,杨小军,李吉明..MIF抑制剂ISO-1通过减轻氧化应激、炎症反应和细胞凋亡缓解脓毒症诱导的急性肾损伤[J].安徽医科大学学报,2024,59(12):2079-2086,8.基金项目
新疆维吾尔自治区自然科学基金资助项目(编号:2020D01C236) Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2020D01C236) （编号:2020D01C236）
