摘要
目的:分析非奈利酮(Finerenone)对大鼠心肌H9C2细胞在脂多糖(LPS)刺激下引起的凋亡和氧化应激的影响及其作用机制。方法:使用不同浓度的非奈利酮(0, 2.5, 5, 10, 20 mmol·L^(-1))经过24 h的处理,接着给予12 h的脂多糖(10μg·mL^(-1))刺激,使用CCK-8试剂盒评估细胞活性,并确定适宜的非奈利酮浓度以进行后续研究。将大鼠心肌H9C2细胞随机分成4组:对照组、仅非奈利酮处理组(浓度为5 mmol·L^(-1))、单独LPS处理组(浓度为10μg·mL^(-1))以及同时接受LPS和非奈利酮处理组(浓度分别为10μg·mL^(-1)和5 mmol·L^(-1))。使用活性氧(ROS)检测试剂盒评估细胞的氧化应激水平,一步法TUNEL细胞凋亡检测试剂盒检测细胞凋亡;Western Blot法检测C-caspase-3、Bax、Bcl-2、PI3K、AKT和GSK-3β表达水平。结果:不同浓度的非奈利酮处理后,对大鼠心肌H9C2细胞的存活率差异无统计意义(P>0.05)。给予非奈利酮处理可挽救由LPS刺激诱导的细胞存活率下降,在非奈利酮浓度为5、10、20 mmol·L^(-1)的组中,差异具有统计学意义(P<0.05)。LPS组细胞凋亡、氧化应激显著高于对照组(P<0.05);非奈利酮处理后能抑制LPS诱导的凋亡因子的表达(P<0.05),同时减轻凋亡和氧化应激的水平(P<0.05)。机制方面,Western Blot显示非奈利酮能激活PI3K/AKT/GSK-3β信号通路。结论:非奈利酮能减轻LPS诱导的细胞毒性,抑制凋亡和氧化应激,而这些作用可能是通过激活PI3K/AKT/GSK-3β信号通路实现的。
Objective:To investigate the mechanism of action of finerenone and its effect on oxidative stress and apoptosis in rat myocardial H9C2 cells stimulated with lipopolysaccharide(LPS).Methods:Fol-lowing a 24-hour course of therapy with varying finerenone concentrations(0,2.5,5,10,and 20 mmol·L^(-1))followed by 12 hours of LPS(10µg·mL^(-1))stimulation,cellular activity was assessed using a CCK-8 kit,and the appropriate finerenone concentration was determined for subsequent studies.Rat myocardial H9C2 cells were randomly divided into four groups:a control group,a finerenone-treated group only(at a concentration of 5 mmol·L^(-1)),an LPS-treated group alone(at a concentration of 10µg·mL^(-1)),and a group that received both LPS and finerenone treatment(at concentrations of 10µg·mL^(-1) and 5 mmol·L^(-1),respectively).Oxidative stress levels in cells were assessed using a reactive ox-ygen species(ROS)assay kit,a one-step TUNEL apoptosis assay kit to assess apoptosis levels;and Western Blot was adopted to detect C-caspase-3,Bax,Bcl-2,PI3K,AKT,and GSK-3βexpression levels.Results:There was no statistically significant difference in the survival of rat myocardial H9C2 cells after treatment with different concentrations of finerenone(P>0.05).Administration of finere-none treatment rescued the decline in cell survival induced by LPS stimulation,and the difference was statistically significant in the groups with finerenone treatment at concentrations of 5,10,and 20 mmol·L^(-1)(P<0.05).When compared to the control group,the LPS group showed noticeably higher levels of oxidative stress and apoptosis.It was discovered that finerenone treatment reduced the amount of apoptotic proteins produced in response to LPS(P<0.05),and at the same time attenuated the levels of apoptosis and oxidative stress(P<0.05).Mechanistically,Western Blot showed that finerenone ac-tivated the PI3K/AKT/GSK-3βsignaling pathway.Conclusion:Finerenone attenuates LPS-induced cytotoxicity and inhibits apoptosis and oxidative stress,and these effects may be achieved by activating the PI3K/AKT/GSK-3βsignaling pathway.
作者
张恒
邓伟
ZHANG Heng;DENG Wei(Dept.of Cardiovascular Medicine,Renmin Hospital of Wuhan University;Hubei Provincial Key Laboratory of Metabolism and Related Chronic Diseases,Wuhan 430060,Hubei,China)
出处
《武汉大学学报(医学版)》
2025年第6期703-708,共6页
Medical Journal of Wuhan University
基金
国家自然科学基金面上项目(编号:82170245
82370284)。
作者简介
张恒,男,1998-,医学硕士,主要从事心血管疾病研究,E-mail:hengzhang98@whu.edu.cn;通信作者:邓伟,男,1982-,医学博士,副教授,主任医师,博士生导师,主要从事心力衰竭与心肌病研究,E-mail:vivideng1982@whu.edu.cn。
