基于ACOX1-CPT2基因表达变化探讨肉苁蓉苯乙醇苷对高脂饮食诱导ApoE-/-小鼠肝组织脂质代谢的改善作用

Study on Improvement Effects of Phenylethanoid Glycosides Extract from Rouchongrong( Cistanche deserticola ) on Lipid Metabolism in Livers of ApoE -/- Mice withHigh Fat Diet Based on Regulating ACOX1-CPT2 Expressions

目的:探讨肉苁蓉苯乙醇苷(phenylethanoid glycosides from Cistanche deserticola, PGCD)对高脂饮食诱导ApoE-/-小鼠肝组织脂质代谢的影响。方法:70只雄性ApoE-/-小鼠随机分为模型组(MOD)、阿伐他汀[20 mg/(kg·d)]+依折麦布[20 mg/(kg·d)]组(A+E)及肉苁蓉苯乙醇苷组[(250、500、1000 mg/(kg·d))](L-PGCD、M-PGCD、H-PGCD),均以高脂饮食(脂肪42%、胆固醇0.15%)饲养,另取14只雄性C57BL/6J小鼠喂食普通饲料作为正常对照组。12周末,检测各组小鼠血浆三酰甘油(TG)含量和谷丙转氨酶(ALT)和谷草转氨酶(AST)活性。取新鲜肝组织匀浆,ELISA方法检测脂肪酸合成酶(FAS)、长链脂酰辅酶A脱氢酶(LCAD)等脂质代谢相关酶的含量;检测谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性及丙二醛(MDA)和过氧化脂质(LPO)含量;RT-PCR检测肝组织酰基辅酶A氧化酶1(ACOX1)和肉碱棕榈酰基转移酶2(CPT2)的基因表达。结果:与模型组相比,3个剂量PGCD均明显降低血清和肝脏TG水平,抑制血清ALT、AST的活性,剂量依赖性地减少FAS含量,显著提升LCAD含量和GSH-Px和SOD活性,并降低MDA和LPO含量。RT-PCR实验显示,PGCD显著升高肝组织ACOX1和CPT2 mRNA表达。结论:肉苁蓉苯乙醇苷能够通过调控肝组织ACOX1和CPT2基因表达改善肝组织脂质代谢,降低肝组织的脂质沉积。

Objective:To explore the effects of Phenylethanoid glycosides extract from Cistanche deserticola(PGCD)on lipids metobolism in ApoE-/-mice liver which induced by high fat diet.Methods:70 male ApoE-/-mice fed with high-fat diet were divided into 5 groups,model(MOD)group,positive control group mice were given Atorvastatin[20 mg/(kg·d)]+Ezetimibe[20 mg/(kg·d)](A+E),PGCD low,middle and high dose groups were given PGCD[250,500,1000 mg/(kg·d)];another 14 male C57BL/6J mice with a chow diet were set as the normal control(CON)group.At the 12 th weekend,the content of TG and the activities of ALT,AST in serum and liver tissue were detected according to the operating instructions.Fresh liver tissue was homogenized and the contents of fatty acid synthase(FAS),long-chain acyl-CoA dehydrogenase(LCAD)and glutathione peroxidase(GSH-Px),superoxide dismutase(SOD)activities were tested by ELISA method,meanwhile,the contents of malondialdehydeand(MDA)and lipid peroxide(LPO)were detected with kid methods.The mRNA expressions of acyl coenzyme A oxidase 1(ACOX1)and carnitine palmitoyltransferase 2(CPT2)were detected by RT-PCR experiments.Results:In contrast to the model group,three-dose PGCD were all reduced the levels of TG in liver and serum with a dose-dependent manner,and lowered the activities of ALT and AST in serum.Three-dose PGCD were all reduced the content of FAS with a dose-dependent fashion,and raised the content of LCAD in liver significantly.PGCD promoted the activities of GSH-Px and SOD and reduced the contents of MDA and LPO in liver significantly.RT-PCR experiments showed that PGCD elevated the mRNA levels of ACOX1and CPT2 in liver tissue.Conclusion:PGCD can ameliorate lipid metabolism and oxidative stress response,reduce lipid deposition in liver tissue by regulating the mRNA expressions of ACOX1 and CPT2.