丹参素异丙酯对肝细胞脂肪过度蓄积的改善作用及其机制  

Study on ameliorative effect of IDHP on excessive fat accumulation in hepatocytes

作　　者：董健 张豫川 苏强 白亚军[3] 孙娜 程燕 郑晓晖[3] DONG Jian;ZHANG Yu-chuan;SU Qiang;BAI Ya-jun;SUN Na;CHENG Yan;ZHENG Xiao-hui(Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Northwest Minzu University,2.College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;The College of Life Sciences,Northwest University,Xi'an 710069,China;Key Lab of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Experimental Center of Northwest Minzu University,Lanzhou 730030,China)

机构地区：[1]西北民族大学生物医学研究中心甘肃省动物细胞技术创新中心,甘肃兰州730030 [2]西北民族大学生命科学与工程学院,甘肃兰州730030 [3]西北大学生命科学学院,陕西西安710069 [4]西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,甘肃兰州730030 [5]西北民族大学实验教学部,甘肃兰州730030

出　　处：《中国药理学通报》2023年第8期1541-1547,共7页Chinese Pharmacological Bulletin

基　　金：甘肃省自然科学基金项目(No 21JR1RA193);西北民族大学引进人才科研项目(No xbmuyjrc2020022)。

摘　　要：目的探索丹参素异丙酯[isopropyl3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate,IDHP]能否通过改善线粒体功能来抑制肝细胞的脂肪蓄积,减轻非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)患者的脂肪过度蓄积症状。方法利用游离脂肪酸棕榈酸和油酸(PA∶OA摩尔比=1∶2)诱导肝细胞脂肪蓄积,模拟细胞脂肪化模型。首先,利用CCK-8检测不同浓度PAOA及IDHP对细胞活力的影响;选用600μmol·L^(-1) PAOA诱导4株肝细胞脂滴蓄积,通过检测尼罗红(Nile Red)荧光信号和甘油三脂(triglycerides,TG)、胆固醇(TC)含量来判定细胞脂肪化模型建立是否成功;分别检测经不同浓度IDHP干预后,细胞内活性氧(ROS)含量、还原型谷胱甘肽(GSH)活性、三磷酸腺苷(ATP)水平及线粒体膜电位(MMP)的变化。结果PAOA诱导后,细胞内出现大量脂滴蓄积,TG、TC含量升高,生成过量的ROS,同时GSH活性降低,ATP合成减少且线粒体膜电位下降。而随着IDHP干预浓度的增加,能有效改善脂滴蓄积,抑制TG的累积,减少活性氧的生成,提高GSH的活性,ATP生成量增加,且线粒体膜电位恢复至正常水平。结论IDHP可通过发挥抗氧化作用缓解氧化应激,改善线粒体功能,调节能量代谢,进而抑制肝细胞脂肪变性。Aim To explore whether isopropyl3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate(IDHP)could inhibit fat accumulation in liver cells by improving mitochondrial function,and alleviate the symptom of excessive fat accumulation in patients with NAFLD.Methods Cell steatosis model was established by inducing hepatocyte fat accumulation using palmitic acid and oleic acid(PA∶OA molar ratio=1∶2).Firstly,the effects of different concentrations of PAOA or IDHP on cell viability were detected by CCK-8.Then 600μmol·L^(-1) PAOA was used to induce lipid droplet accumulation in four hepatocytes.The fluorescence signal of Nile Red and the content of triglyceride(TG)and cholesterol(TC)were detected to determine whether the cell fatty transformation model was successfully established.After that intracellular reactive oxygen species(ROS)content,glutathione(GSH)activity,adenosine triphosphate(ATP)level,and changes in mitochondrial membrane potential(MMP)were detected after intervention with IDHP.Results With PAOA induction,a large number of lipid droplets accumulated in cells,the content of TG and TC increased,and excess ROS was generated.While the GSH activity was reduced,both ATP content and mitochondrial membrane potential were also reduced significantly.After the intervention of IDHP with different concentrations,both lipid droplets and TG content could be effectively reduced,so could the production of ROS.The activity of GSH was significantly promoted,and even more,the content of ATP apparently increased.Besides,the mitochondrial membrane potential could be effectively recovered to normal level.Conclusions IDHP could efficiently inhibit hepatocyte steatosis through exhibiting antioxidant effect.Furthermore,it could also improve mitochondrial function by regulating energy metabolism.

关 键 词：PAOA 肝细胞脂肪化 非酒精性脂肪性肝病 IDHP 线粒体膜电位 线粒体功能 

分 类 号：R284.1[医药卫生—中药学] R322.47[医药卫生—中医学] R329.24R344.2R575.5