摘要
目的观察水飞蓟宾对油酸诱导的非酒精性脂肪肝(NAFLD)模型脂代谢的影响。方法将细胞分为空白组、模型组(0.2 mmol·L^(-1)油酸)和3个剂量实验组(0.2 mmol·L^(-1)油酸+2.5,5.0,10.0μg·m L^(-1)水飞蓟宾)。以酶偶联比色法检测细胞三酰甘油(TG),以黄嘌呤氧化酶法测定超氧化物歧化酶(SOD),以硫代巴比妥酸法测定丙二醛(MDA),以酶促法测定谷胱甘肽过氧化物酶(GSH-Px),以酶联免疫吸附法测定肿瘤坏死因子-α(TNF-α)水平,以实时荧光定量PCR法检测细胞腺苷酸活化蛋白激酶(AMPK)、去乙酰化酶(Sir T3)、乙酰辅酶A羧化酶2(ACC 2)及肉碱棕榈酰转移酶^(-1)A(CPT^(-1)A)的mRNA表达水平,以油红O染色观察细胞内脂滴沉积变化。结果高剂量实验组TG水平为(164.74±53.94)mg·g^(-1),与模型组的(500.05±152.01)mg·g^(-1)相比显著降低(P<0.01);与模型组SOD水平(4.96±1.39)U·mg^(-1)比较,高剂量实验组SOD水平(20.37±11.16)U·mg^(-1)显著增高(P<0.01)。与模型组MDA、TNF-α水平(0.20±0.03)U·mg^(-1)、(32.03±8.04)pg·m L^(-1)比较,高剂量实验组的MDA、TNF-α水平(0.10±0.02)U·mg^(-1)、(15.00±0.53)pg·m L^(-1)显著降低(P<0.01,P<0.05)。与模型组比较,高剂量实验组的Ampk、Sir T3、CPT^(-1)A mRNA表达水平显著增加(P<0.01),而ACC2 mRNA表达显著降低(P<0.01)。结论水飞蓟宾有效降低油酸诱导的NAFLD模型肝细胞TG水平,可能与其抗氧化、调节脂代谢相关基因表达水平相关。
Objective To observe the effects of silibinin on the lipid metabolism of cellular model of non- alcoholic fatty liver disease（ NAFLD） induced by oleic acid. Methods The cellular models of NAFLD was divided into normal group, model group（ OA 0. 2mmol·L^-1）,high dosage,moderate dosage and low dosage test groups（ OA 0. 2 mmol·L^-1＋ silibinin 2. 5,5. 0,10. 0 μg·mL^-1,respectively）.The cellular levels of triglyceride（ TG） were determined by enzyme coupling chromometry. The superoxide dismutase（ SOD） were measured by xanthine oxidase reaction. The malonaldehyde（ MDA） were evaluated by thiobarbituric acid method. The glutathione peroxidase（ GSH- PX） were determined by enzymatic method. The tumor necrosis factor- α（ TNF- α） were detected by ELISA. The adenosine monophosphate activated protein kinase（ AMPK）,Sirtuin- 3（ SirT 3）,acetyl- Co A carboxylase 2（ ACC 2） and carnitine palmitoyltransferase- 1A（ CPT- 1A） mRNA expression were quantitatively determined byreal- time quantitative PCR（ RT- PCR）. The intracellular lipid acceleration was observed by oil red O stain.Results Compared to the model group on TG level with（ 500. 05 ± 152. 01） mg·g^-1,TG level in high dosage test group with（ 164. 74 ± 53. 94） mg·g^-1decreased significantly（ P〈0. 01）. Compared to the model group on SOD level with（ 4. 96 ± 1. 39） U·mg^-1,the SOD level in high dosage test group with（ 20. 37 ± 11. 16） U·mg^-1was increased significantly（ P〈0. 01）. Compared to the model group on MDA and TNF- α levels with（ 0. 20 ± 0. 03） U·mg^-1,（ 32. 03 ± 8. 04） pg ·mL^-1,MDA and TNF- α levels in high dosage test group with（ 0. 10 ± 0. 02） U·mg^-1,（ 15. 00 ± 0. 53） pg·mL^-1were decreased significantly（ P〈0. 01,P〈0. 05）. Compared to the model group,Ampk,SirT 3,CPT- 1A mRNA expression levels in high dosage test group were much higher（ P〈0. 01）,but ACC2 mRNA expression level was much lower（ P〈0. 01）. Conclusion The TG level of cellular model of NAFLD could be decreased effectively by silibinin,which might be related to its antioxidant activity and regulation of lipid metabolism related gene expression levels.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2016年第13期1202-1205,共4页
The Chinese Journal of Clinical Pharmacology
基金
新疆维吾尔自治区高校科研计划基金资助项目(XJEDU2014I020)
国家自然科学基金资助项目(81360669)
关键词
水飞蓟宾
非酒精性脂肪肝
瘤坏死因子-α
silibinin
non-alcoholic fatty liver disease
tumor necrosis factor-α
