小檗碱通过抑制脂肪酸摄取降低小鼠原代肝细胞脂质沉积

小檗碱通过抑制脂肪酸摄取降低小鼠原代肝细胞脂质沉积俞牧雨;米日阿依·阿里木江;刘威;殷峻
【期刊名称】《中国药理学通报》
【年(卷),期】2018(034)003
【摘要】Aim To investigate the role of berberine in mouse primary hepatocytes steatosis and whether aden-osine monophosphate-activated protein kinase(AMPK) is essential in this process in order to explore the mech-anism of non-alcoholic fatty liver disease treatment. Methods Different concentrations of oleic acid(OA) were used in mouse primary hepatocytes to determine the appropriate dose inducing steatosis. Subsequently, hepatocytes were treated with berberine and OA at the same time for 24 h serving metformin as positive con-trol. Lactic dehydrogenase (LDH) release test was performed to investigate cell viability. Lipid level was determined by oil red staining and triglyceride assay. Western blot measured the phosphorylation level of AMPK and Acetyl CoA carboxylase. An AMPK inhibi-tor compound C(CC) pre-treated hepatocytes for 1 h followed by berberine 24 h-treatment. The relationship between free fatty acid(FFA) uptake and mitochondri-al inhibition was evaluated by measuring FFA in the supernatant of OA,berberine and rotenone (mitochon-drial complex I inhibitor) group. Results Berberine could significantly reduce primary hepatocytes steatosis induced by oleic acid and stimulate AMPK and ACC phosphorylation at a non-toxic dose. In
addition, CC obviously inhibited AMPK activity,but failed to dimin-ish the lipid dysregulation improvement of berberine. Berberine and rotenone intervention reduced OA up-take by 31.2% and 23.6%,respectively. Conclusion Berberine ameliorates hepatocytes lipid accumulation by suppressing fatty acid uptake,which is probably re-sulted from inhibition of mitochondrial respiratory chain complex I,independently of AMPK activation.%目的观察小檗碱对小鼠原代肝细胞脂质沉积的影响及其是否依赖于磷酸腺苷活化蛋白激酶(adenosine 5′-mono-phosphate-activated protein kinase,AMPK)通路,探讨小檗碱治疗非酒精性脂肪肝的作用机制.方法用不同浓度白蛋白偶联的油酸(OA)孵育小鼠原代肝细胞,确定OA诱导脂肪变的具体浓度,同时以二甲双胍为阳性对照,给予小檗碱处理24 h,乳酸脱氢酶(LDH)试剂盒检测确定小檗碱的安全剂量;油红染色定性判断细胞内脂质水平;酶法定量检测细胞内甘油三酯含量;Western blot法检测AMPK以及乙酰辅酶A羧化酶(ACC)磷酸化水
平.AMPK 的特异性阻断剂Compound C(CC)孵育原代肝细胞1 h后,再用小檗碱处理细胞24 h,油红染色及甘油三酯检测判断AMPK被抑制时小檗碱对脂质沉积的作用.生化分析仪检测 OA 组、小檗碱组、鱼藤酮(线粒体复合物I抑制剂)组细胞上清液中脂肪酸的含量,观察细胞对脂肪酸的摄取情况与复合物I抑制之间的关系.结果小檗碱在无细胞毒性的剂量下可明显降低小鼠原代肝细胞中油酸所致脂质沉积.同时,小檗碱可使AMPK及ACC 的磷酸化水平增加.此外,在加入 CC 即AMPK 活性被抑制的情况下,小檗碱降低肝细胞脂质沉积的作用不受影响.小檗碱和鱼藤酮分别使肝细胞的油酸摄取减少约31.2%和23.6%.结论小檗碱通过抑制电子传递链线粒体复合物I抑制脂肪酸摄取,进而降低小鼠原代肝细胞脂质沉积,这一作用并不依赖于AMPK通路.
【总页数】6页(P337-342)
【作者】俞牧雨;米日阿依·阿里木江;刘威;殷峻
【作者单位】上海交通大学附属第六人民医院内分泌代谢科,上海市糖尿病研究所,上海市糖尿病重点实验室,上海市糖尿病临床医学中心,上海市代谢病临床医学中心;上海交通大学附属第六人民医院内分泌代谢科,上海市糖尿病研究所,上海市糖尿病重点实验室,上海市糖尿病临床医学中心,上海市代谢病临床医学中心;上海交通大学附属第六人民医院骨科,上海 200233;上海交通大学附属第六人民医院内分泌代谢科,上海市糖尿病研究所,上海市糖尿病重点实验室,上海市糖尿病临床医学中心,上海市代谢病临床医学中心
【正文语种】中文
【中图分类】R-332;R284.1;R329.24;R345.57;R575.502.2;R977.6
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