芦丁通过调控TG代谢途径抑制肝细胞脂肪变性的作用机制研究  

Rutin Inhibits Oleic Acid Induced Lipogenesis in Hepatocyte Cells via Regulating TG Metabolic Pathway

作　　者：潘然[1] 丁滨[1] 胡林峰[1] 张福利[1] 丁蕾[1] 方波[1] 谢璐帆 窦晓兵[1] 

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《云南中医学院学报》2015年第5期1-6,共6页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81473393;81241145);浙江省自然科学基金(LY12C07002)

摘　　要：目的通过体外建立油酸(Oleic acid,OA)诱导的脂肪变性肝细胞Hep G2模型探索芦丁预防脂肪聚积的分子机制。方法利用乳酸脱氢酶(LDH)释放评价芦丁对Hep G2细胞毒性的影响;用0.5 mmol/L OA诱导人肝细胞株Hep G2建立体外脂肪聚积细胞模型,并通过油红O染色观察和甘油三酯(TG)含量判定细胞模型是否建立成功;分别采用实时荧光定量PCR技术及Western Blot技术在转录水平及翻译水平检测相关蛋白的表达差异。结果芦丁和油酸不会导致Hep G2细胞中LDH激增,但油酸会造成细胞内TG聚积。而芦丁能有效抑制OA诱导的Hep G2细胞中TG的聚积,以及二脂酰甘油酰基转移酶(Diacylgycerola cyltransferase,DGAT)的转录(P<0.05),同时激活过氧化物酶体增殖物激活受体α(Peroxisome proliferator-activated receptor alpha,PPAR-α)的转录(P<0.05)及翻译(且与芦丁浓度计量相关),进而促进其靶基因肉毒碱棕榈酰转移酶(Carnitine palmitoyltransferase,CPT)的转录。结论芦丁能够激活脂肪变性肝细胞Hep G2中PPAR-α的转录和翻译,进而促进其靶基因CPT的转录,加剧了脂肪酸的降解代谢,与此同时对DGAT转录的抑制阻止了TG的合成。该结果进一步阐明了芦丁防治非酒精性脂肪性肝病的作用机制,为芦丁的临床使用提供了理论依据和实验基础。Objective To study the role of rutin in TG accumulation in oleic acid induced hepatocyte cells. Methods The effect of rutin and OA induced cell death was determined by lactate dehydrogenase（LDH）release assays. Human liver cell line Hep G2 was used to establish the cell model by 0.5mmol/L OA. We measured TG level and use oil red O stain to determine whether cell model is successful or not. The level of proteins adducts were determined by Western blotting. The level of gene adducts were determined by Realtime PCR. Results Rutin and oleic acid could not increased LDH surge,but cause the oleic acid induced intracellular TG accumulation in Hep G2 cells. Rutin could inhibit TG accumulation and DGAT expression in Hep G2 cells,increasing the expression of PPAR-α and CPT. Conclusion Rutin activates the transcription and translation of PPAR-α in fatty degeneration of Hep G2,and promotes the transcription of its target gene CPT and fatty acid degradation. It also prevents the transcription of DGAT,and inhibits oleic acid induced TG accumulation. The results elucidate the prevention and treatment of rutin in nonalcoholic fatty liver disease,provide theoretical and experimental basis for the clinical use of rutin.

关 键 词：油酸 肝细胞 芦丁 二脂酰甘油酰基转移酶 肉毒碱棕榈酰转移酶 

分 类 号：R285.5[医药卫生—中药学]