新疆小枝玫瑰醇提物对脂肪酸诱导的HepG2细胞脂质蓄积的改善作用

Effect of Alcohol Extract of Xinjiang Ramulus Rose on Fatty Acid Induced Lipid Accumulation in HepG2 Cells

摘要:

为探究新疆小枝玫瑰醇提物对HepG2细胞脂肪堆积的影响，以新疆小枝玫瑰醇提物为对象，用脂肪酸（free fatty acids，FFAs）诱导HepG2细胞建立体外脂肪酸负荷模型。MTT法筛选小枝玫瑰对HepG2细胞的安全浓度。用甘油三酯（triglyceride，TG）试剂盒测定小枝玫瑰醇提物干预后各组细胞的TG含量。油红O染色观察并测定各组细胞内脂滴蓄积情况和脂质含量。Western blot法测定各组细胞过氧化物酶体增殖物激活受体α（peroxisome proliferator-activated receptor alpha，PPARα）、酰基辅酶A氧化酶1（peroxisomal acyl-coenzyme aoxidase 1，ACOX1）、肉毒碱棕榈酰基转移酶1A（carnitine palmitoyltransferase 1A，CPT1A）蛋白水平。结果显示：小枝玫瑰醇提物作用HepG2细胞的安全浓度≤200 μmol/L。以FFAs浓度为1.5 mmol/L孵育24 h可成功诱导HepG2细胞体外脂肪酸负荷模型。与模型组相比，小枝玫瑰醇提物组呈剂量依赖性地显著（p<0.05）降低FFAs诱导的细胞内TG和脂质含量，TG水平下降47.17%~60.38%，脂质含量下降25.00%~36.61%；与模型组相比，小枝玫瑰醇提物能显著上调PPARα及其下游ACOX1、CPT1A蛋白表达水平（p<0.05）。小枝玫瑰醇提物可以改善FFAs诱导的HepG2细胞脂质蓄积，其抗脂变作用是通过激活PPARα介导的脂肪酸氧化代谢通路，进而减少细胞内TG水平和脂质含量实现的。

Abstract:

To explore the influence of the alcohol extract of Xinjiang Branchlets Rosa rugosa Thunb. on the accumulation of fat in HepG2 cells, the alcohol extract of Branchlets Rosa rugosa Thunb. was used as the object, HepG2 cells were induced with free fatty acids (FFAs) to establish a fatty acid load model in vitro. The safe concentration of Branchlets Rosa rugosa Thunb. in the HepG2 cells was determined by MTT method. The triglyceride (TG) kit was used to measure the TG content of each group of cells after the intervention of the alcohol extract of Branchlets Rosa rugosa Thunb. Intracellular lipid droplet accumulation and lipid content of each group were determined by oil red O staining. Western blotting method was used to detect the protein levels of PPARα, ACOX1, and CPT1A in each group. The results showed that the safe concentration of alcohol extract of Branchlets Rosa rugosa Thunb. in HepG2 cells was equal to or less than 200 μmol/L. The fatty acid load model of HepG2 cells in vitro could be successfully established with FFAs concentration of 1.5 mmol/L for 24 h. Compared with the model group, the ethanol extract of Branchlets Rosa rugosa Thunb. markedly decreased the intracellular TG and lipid content induced by FFAs in a dose-dependent manner (p<0.05). The TG level decreased by 47.17%~60.38%, and the lipid content decreased by 25.00%~36.61%. Compared with the model group, the ethanol extract of Branchlets Rosa rugosa Thunb. could obviously up-regulate PPARα, ACOX1and CPT1A protein expression (p<0.05). The ethanol extract of Branchlets Rosa rugosa Thunb. can improve FFAs-induced lipid accumulation in HepG2 cells, and its anti-lipidemia effect isachieved by activating PPARα-mediated fatty acid oxidation metabolism pathway, and then reducing intracellular TG and lipid content.