苦丁冬青苷D抑制肝细胞脂质沉积作用及机制OA北大核心CSTPCD
Inhibition effect of kudinoside D on lipid deposition in hepatocytes and its mechanism
目的 探讨苦丁冬青苷D(kudinoside D,KD-D)对棕榈酸(palmitic acid,PA)诱导肝细胞脂质沉积的影响.方法 体外培养AML-12细胞,随机分为Control组、PA组、PA+KD-D 20 μmol·L-1 组、PA+KD-D 40 μmol·L-1 组和 PA+KD-D 80 μmol·L-1组.除Control组外,其他各组细胞加入PA(0.4 mmol·L-1)孵育 24 h,KD-D 在 PA 加入前 1 h 给予.MTT法检测细胞存活率;油红O染色和透射电子显微镜检测肝细胞脂质沉积;DCFH-DA荧光探针检测细胞内活性氧族;MitoSOX线粒体超氧化物红色荧光探针检测细胞线粒体超氧化物.结果 不同浓度KD-D明显改善PA诱导的肝细胞形态学改变.与Control组比较,PA组细胞内红色脂滴明显增加;与PA组比较,KD-D不同浓度干预的细胞内红色脂滴减少.透射电子显微镜结果提示,KD-D减少PA诱导的肝细胞脂肪变性并改善超微结构.此外,KD-D明显降低PA诱导的细胞活性氧水平(P<0.01),降低细胞线粒体超氧化物含量(P<0.01).结论 KD-D可抑制PA诱导的肝细胞脂质沉积,其机制可能与调控氧化应激反应有关.
Aim To investigate the effect of kudinoside D(KD-D)on palmitic acid(PA)-induced lipid depo-sition in hepatocytes.Methods Mouse hepatocytes AML-12 were cultured and randomly divided into the Control group,PA group,PA+KD-D 20 μmol·L-1 group,PA+KD-D 40 μmol·L-1 group and PA+KD-D 80 μmol·L-1 group.AML-12 cells in PA and KD-D groups were treated with PA(0.4 mmol·L-1)for 24 h.AML-12 cells in KD-D groups were incubated with KD-D for 1 h before stimulation with PA.MTT as-say was used to detect cell survival rate,oil red O stai-ning and transmission electron microscopy were used to detect lipid deposition in cells,DCFH-DA fluorescence probe was used to detect intracellular reactive oxygen species(ROS)and MitoSOX mitochondrial superoxide red fluorescence probe was used to detect mitochondrial superoxide content in cells.Results KD-D at differ-ent concentrations improved PA-induced changes in cell morphology significantly.Compared with the Con-trol group,cells in PA group showed a significant in-crease in intracellular lipid droplets.Compared with PA group,the red lipid droplets in KD-D groups de-creased.The results of transmission electron microsco-py demonstrated that KD-D reduced PA-induced hepat-ic steatosis and improved ultrastructure.In addition,KD-D significantly decreased PA-induced cellular ROS level(P<0.01)and reduced mitochondrial superox-ide content(P<0.01).Conclusion KD-D inhibits PA-induced lipid deposition by regulating the cellular oxidative stress levels in AML-12 cells.
薛彩彩;厉彦翔;乔秀梅;彭金咏;王金红
山东第二医科大学药学院"分子药理学与转化研究"山东省高校重点实验室,山东潍坊 261053山东第二医科大学药学院"分子药理学与转化研究"山东省高校重点实验室,山东潍坊 261053||大连医科大学药学院,辽宁大连 116044
中医学
苦丁冬青苷DAML-12细胞棕榈酸脂质沉积氧化应激非酒精性脂肪性肝病
kudinoside DAML-12 cellspalmitic acidlipid depositionoxidative stressnon-alcoholic fatty liver disease
《中国药理学通报》 2024 (009)
1688-1694 / 7
山东省自然科学基金资助项目(No ZR2021QH095);潍坊医学院附属医院科技发展项目重点项目(No 2023FYZ002)
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