摘要
目的:观察人参皂苷Rd对肝细胞Derlin-1/PERK/eIF2α/CHOP信号通路的影响,从而阐明其调控内质网应激治疗非酒精性脂肪性肝炎(NASH)的分子机制。方法:采用CCK8法检测人参皂苷Rd的肝细胞毒性,随后通过游离脂肪酸诱导HepG2构建NASH细胞模型,并给予人参皂苷Rd和内质网抑制剂进行干预。分别采用油红O染色、生化检测、免疫荧光、酶联免疫吸附试验、蛋白印迹法和实时荧光定量PCR检测人参皂苷Rd对甘油三酯、肝功能、氧化应激、细胞因子和Derlin-1/PERK/eIF2α/CHOP信号通路中关键分子表达的影响。结果:与模型组相比,人参皂苷Rd能够显著抑制HepG2细胞中TG的含量和AST与ALT的水平,增加GSH的含量和SOD的活性,降低MDA的含量和ROS的平均荧光强度,差异均具有统计学意义(P<0.01)。此外,人参皂苷Rd能够显著抑制HepG2细胞中TNF-α、IL-1β、CCL2和CXCL1的含量,下调ATF4、CHOP蛋白的表达和降低p-PERK/PERK和p-eIF2α/eIF2α的比值,与模型组比较,差异均具有统计学意义(P<0.01)。不仅如此,人参皂苷Rd干预还能显著抑制Derlin-1蛋白和mRNA的表达,与模型组比较,差异均具有统计学意义(P<0.01)。结论:人参皂苷Rd通过抑制Derlin-1/PERK/eIF2α/CHOP信号通路的激活,从而抑制内质网应激,达到治疗NASH的作用。
Objective:To investigate the effects of ginsenoside Rd on the Derlin-1/PERK/eIF2α/CHOP signaling pathway in hepatocytes,thereby elucidating its molecular mechanism in regulating endoplasmic reticulum stress for the treatment of non-alcoholic steatohepatitis(NASH).Methods:The cytotoxicity of ginsenoside Rd was assessed using the CCK8 assay.Subsequently,a NASH cell model was established by inducing HepG2 cells with free fatty acids,followed by intervention with ginsen-oside Rd and an endoplasmic reticulum inhibitor.The effects of ginsenoside Rd on triglyceride(TG)levels,liver function,oxidative stress,cytokines,and the expression of key molecules in the Derlin-1/PERK/eIF2α/CHOP signaling pathway were evaluated using Oil Red O staining,biochemical assays,immunofluorescence,enzyme-linked immunosorbent assay(ELISA),Western blotting,and real-time quantitative PCR.Results:Compared to the model group,ginsenoside Rd significantly reduced TG content and the levels of AST and ALT in HepG2 cells,increased GSH content and SOD activity,and decreased MDA content and ROS mean fluorescence intensity,with statistically significant differences(P<0.01).Additionally,ginsenoside Rd significantly inhibited the levels of TNF-α,IL-1β,CCL2,and CXCL1 in HepG2 cells,downregulated the expression of ATF4 and CHOP proteins,and reduced the ratios of p-PERK/PERK and p-eIF2α/eIF2α,with statistically significant differences compared to the model group(P<0.01).Moreover,ginsenoside Rd intervention significantly suppressed the expression of Derlin-1 protein and mRNA,with statistically significant differences compared to the model group(P<0.01).Conclusion:Ginsenoside Rd inhibits the activation of the PERK/eIF2α/CHOP signaling pathway,thereby suppressing endoplasmic reticulum stress and exerting therapeutic effects on NASH.
作者
路瑶
李丽萍
王炳予
袁星星
LU Yao;LI Liping;WANG Bingyu;YUAN Xingxing(Graduate School,Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150036,China;Department of Endoscopy,Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150006,China;Department of Gastroenterology,Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150006,China)
出处
《海南医科大学学报》
北大核心
2025年第16期1220-1228,共9页
Journal of Hainan Medical University
基金
黑龙江省自然科学基金优秀青年项目(YQ2022H015)。
