摘要
目的:探究鲜地黄中两个酰胺类化合物rehmagluamide(Reh)和rehmanalkaloid C(Reh C)对LPS诱导NRK-52e细胞损伤的干预作用及潜在机制。方法:建立LPS诱导的NRK-52e细胞损伤模型,加入SIRT3选择性抑制剂3-TYP,ELISA法检测细胞上清液中炎症因子TNF-α与IL-6水平;流式细胞术检测细胞凋亡、活性氧(ROS)与线粒体膜电位(JC-1)水平;细胞免疫荧光检测细胞中SIRT3蛋白及线粒体凋亡关键蛋白Caspase-3、Cleaved Caspase-9、Bax、Bcl-2表达。结果:与模型组比较,Reh和Reh C可不同程度降低LPS作用下NRK-52e细胞凋亡及TNF-α、IL-6、ROS水平和Caspase-3、Cleaved Caspase-9蛋白表达,升高线粒体膜电位、SIRT3蛋白表达及Bcl-2/Bax水平,但在加入3-TYP后Reh和Reh C的改善作用显著降低。结论:Reh和Reh C可能通过SIRT3/Bcl-2/Caspase-3信号通路减轻LPS诱导的NRK-52e细胞损伤。
Objective:To investigate the effect and potential mechanism of two amides rehmagluamide(Reh)and rehmanalkaloid C(Reh C)in Fresh Rehmanniae Radix on LPS-induced NRK-52e cell injury.Methods:The LPS-induced NRK-52e cell injury model was established,SIRT3 selective inhibitor 3-TYP was added,and the levels of inflammatory cytokines TNF-αand IL-6 in cell supernatant were detected by ELISA.Apoptosis,reactive oxygen species(ROS)and mitochondrial membrane potential(JC-1)levels were detected by flow cytometry.The expressions of SIRT3 protein and key proteins of mitochondrial apoptosis Caspase-3,Cleaved Caspase-9,Bax and Bcl-2 were detected by immunofluorescence assay.Results:Compared with the model group,Reh and Reh C reduced the cell apoptosis and levels of TNF-α,IL-6,ROS and Caspase-3,Cleaved Caspase-9 proteins expressions in LPS-induced NRK-52e cells to varying degrees,increased mitochondrial membrane potential,SIRT3 protein expression and Bcl-2/Bax level.However,the improvement of Reh and Reh C decreased significantly after the addition of 3-TYP.Conclusion:Reh and Reh C may alleviate LPS-induced NRK-52e cell damage through SIRT3/Bcl-2/Caspase-3 signaling pathway.
作者
刘萌
曾梦楠
阚玉璇
刘晏灵
冯卫生
郑晓珂
LIU Meng;ZENG Meng-nan;KAN Yu-xuan;LIU Yan-ling;FENG Wei-sheng;ZHENG Xiao-ke(Henan University of Chinese Medicine,Zhengzhou 450046,China;Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046,China)
出处
《中药材》
CAS
北大核心
2022年第5期1212-1217,共6页
Journal of Chinese Medicinal Materials
基金
国家重点研发计划-中医药现代化研究专项(2019YFC1708800,2017YFC1702800)
河南省重大科技专项(171100310500)
河南省高层次人才特殊支持计划“中原千人计划”-中原领军人才(ZYQR201810080)
