(-)-羟基柠檬酸调控鸡胚原代肝细胞脂滴沉积的机制  

Mechanisms of (-)-hydroxycitric regulated lipid droplet deposition in primary chicken hepatocytes

作　　者：李龙龙[1] 彭梦玲 王典[1] 俞磊[1] 葛重阳 马海田[1] 

机构地区：[1]南京农业大学农业部动物生理生化重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2017年第3期514-520,共7页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31572483);江苏省自然科学基金项目(BK20151434)

摘　　要：[目的]本文旨在研究(-)-羟基柠檬酸[(-)-HCA]对鸡胚原代肝细胞脂滴沉积的影响及其机制。[方法]选用含(-)-HCA的培养液孵育鸡胚原代肝细胞后,油红染色法检测鸡胚原代肝细胞中脂滴的数量和总面积;试剂盒法检测鸡胚原代肝细胞中甘油三酯、葡萄糖含量及糖代谢、呼吸链关键酶活性。[结果](-)-HCA处理显著降低鸡胚原代肝细胞中甘油三酯含量、脂滴数量和总面积,但对细胞活力和死亡率无显著影响。(-)-HCA处理提高了鸡胚原代肝细胞中葡萄糖消耗量,显著降低了柠檬酸裂解酶活性和细胞质中乙酰辅酶A含量。(-)-HCA处理显著提高了鸡胚原代肝细胞中葡萄糖激酶、磷酸果糖激酶1、丙酮酸激酶、丙酮酸脱氢酶(E1)、乌头酸酶、琥珀酸脱氢酶、苹果酸脱氢酶、柠檬酸合酶、NADH脱氢酶和ATP合酶活性。[结论](-)-HCA通过抑制柠檬酸裂解酶的活性减少细胞质中乙酰辅酶A的供给,并通过增强糖代谢和呼吸链中关键酶活性而加速能量代谢,最终抑制了鸡胚原代肝细胞中脂滴的沉积。[ Objectives ] This study was aimed to investigate the effects of （-）-hydroxycitric [ （-）-HCA] on the lipid droplet deposi- tion and potential biochemistry mechanism in primary chicken embryo hepatocytes. [ Methods] Primary chicken embryo hepatocytes were exposed to（-） -HCA treatment. Oil red staining was used to detect the count and area of lipid droplets in hepatocytes. In addi- tion,the contents of triglyceride, glycogen and glucose in hepatocytes and the activities of glucose metabolism and respiratory chain related enzymes were determined. [ Results] No significant difference was observed on the cell viability and cell death rate in primary chicken hepatocytes after （-）-HCA treated. Oil red O staining analysis showed （-）-HCA significantly decreased the number and total area of lipid droplets, meanwhile the triglyceride content significantly decreased in （-）-HCA treated group compared with that in the control group. （-）-HCA treatment significantly increased the glucose consumption in primary chicken hepatocytes, while the ATP-citrate lyase activity and acetyl coenzymy A （ CoA ） content in cytosol significantly decreased in （ - ） -HCA treated group. Compared with control group, （-）-HCA promoted the activities of glucokinase, phosphofructokinase-1, pyruvate kinase, pyruvate dehydrogenase（ E1 ）, citrate synthase, aconitase, succinate dehydrogenase, malate dehydrogenase, NADH dehydrogenase and ATP synthase remarkably. [ Conclusions] （-）-HCA decreased lipid droplets accumulation by reducing the acetyl CoA supply, which was mainly achieved via inhibition of ATP-citrate lyase activity, and accelerating the energy metabolism in primary chicken hepatocytes.

关 键 词：(-)-羟基柠檬酸 脂代谢 葡萄糖代谢 能量代谢 鸡胚原代肝细胞 

分 类 号：S852.23[农业科学—基础兽医学]