摘要
目的:观察7周有氧运动联合黑果枸杞色素补充对非酒精性脂肪肝病(NAFLD)小鼠肝脏脂肪酸氧化相关因子表达、线粒体氧化速率及抗氧化能力的影响,探讨有氧运动和补充黑果枸杞色素在NAFLD良性转归中的干预作用、交互效应及可能机制。方法:3周龄KM种雄性小鼠48只,随机分为空白组(n=8)和高糖高脂组(n=40)。高糖高脂组采用高糖高脂膳食灌服7周建立NAFLD模型。建模成功后,随机分为模型组(MX组)、有氧运动组(E组)、黑果枸杞色素组(C组)和有氧运动联合黑果枸杞色素组(EC组),E、EC组的有氧运动干预采用中等强度跑台运动,坡度0°,速度12 m/min,时间60 min,每周6次,共7周。C、EC组每日补充黑果枸杞色素(200 ml/kg体重),MX、E组灌服同等体积生理盐水,共7周。干预结束后,测定体重、肝湿重、血脂水平、血清转氨酶活性,肝组织切片观察肝脏组织结构,ELISA检测肝脏TG含量,比色法监测线粒体β氧化速率、TARBS、GSH-px含量,羟胺法测定SOD活性,SABC法检测肝脏PPARα、TNFα蛋白阳性表达水平,RT-PCR检测肝脏PPARα、TNFα、LCAD、CPT1、AOX、CYP4A10、CYP4A12 mRNA表达。结果:(1)镜下显示,MX组小鼠肝脏结构紊乱,细胞核畸形变位于细胞边缘,肝细胞中出现大量脂滴空泡样变,各干预组肝索较清晰,肝细胞脂滴空泡样变减少,细胞核圆居中,脂质变性程度下降。(2)与MX组相比,E、EC组体重、肝湿重下降(P<0.01),C组体重无显著性差异,肝湿重下降(P<0.01);E、C、EC组血清TG、TC、LDL-C、ALT、AST水平下降(P<0.01或P<0.05),HDL-C水平升高(P<0.01);肝脏TG、TBARS含量下降(P<0.01),GSH-px、SOD含量、肝线粒体β氧化速率上升(P<0.01或P<0.05)。对于上述血清和肝脏检测指标,除血清AST水平以外,有氧运动与黑果枸杞色素间均存在协同效应。(3)与MX组相比,E、C、EC组肝脏脂肪酸氧化调控基因TNFα基因表达、蛋白阳性表达水平下降,PPARα基因表达、蛋白阳性表达水平升高(P<0.05或P<0.01),下游靶基因AOX、CPT1、LCAD、CYP4A10、CYP4A12mRNA表达上调(P<0.05)。对于PPARαmRNA表达、TNFα蛋白阳性表达、CPT1 mRNA表达、CYP4A10mRNA表达,有氧运动与黑果枸杞色素存在协同效应。结论:7周有氧运动及黑果枸杞色素补充可明显改善NAFLD小鼠肝脏脂质代谢紊乱,可能通过促进肝脏脂肪酸氧化减少肝细胞中脂质蓄积,降低肝脏脂质变性程度,同时降低肝脏氧化损伤,促进非酒精性脂肪肝的良性转归,有氧运动与黑果枸杞色素联合应用效果最佳。
Objective To observe the effect of 7-week aerobic exercise combined with pigment of lycium ruthenicum murr supplement on the expression of lipid synthesis related genes,mitochondrial oxidation rate and antioxidant capacity in non-alcoholic fatty liver disease(NAFLD)mice,so as to explore the intervention and interaction of aerobic exercise combined with pigment of lycium ruthenicum murr administration on the positive outcome of NAFLD and its mechanism.Methods Forty-eight 3-week-old male Kunming(KM)mice were randomly divided into a control group(n=8)and an experimental group(n=40).The experimental group was successfully induced NAFLD using a high-fat and high-sugar diet for 7 weeks,and randomly divided into a model(MX)group,an aerobic exercise(E)group,a pigment of lycium ruthenicum murr(C)group and an aerobic exercise combined with pigment of lycium ruthenicum murr(EC)group,each of 8.Group E and EC were trained using the treadmill running(0,12 m/min,60 min/d,6/w)for 7 weeks.Group C and EC were given pigment of lycium ruthenicum murr at a dose of 200 mg/kg body weight per day,while group MX and E were given the same amount of normal saline.After the intervention,the Body weight,liver wet weight,serum levels of lipid and aminotransferase were detected.Liver tissue slices were prepared to observe the histopathological changes.The content of triglyce-ride(TG)in liver was measured using the enzyme-linked immunosorbent assay(ELISA),the rate of mitochondrial beta oxidation,content of glutathione peroxidase(GSH-px)and thiobarbituric acid reactive substance(TARBS)were evaluated using the colorimetry,the activity of superoxide dismutase(SOD)was determined using the hydroxylamine.Moreover,the expression of peroxisome proliferator-activated receptorsα(PPARα)and tumor necrosis factor-α(TNFα)was detected using the SABC,while the mRNA levels of PPARα,TNFα,long-chain acyl-CoA dehydrogenase(LCAD),carnitine palmitoyltransferaseⅠ(CPT1),acetyl-coenzyme A oxidase(AOX),cytochrome P450,family 4,subfamily A,polypeptide 10(CYP4 A10),cytochrome P450 and family 4,subfamily A,polypeptide 12(CYP4 A12)were found using the reverse-transcription-polymerase chain reaction(RT-PCR).Results The microscopic structure showed that in group MX,the hepatic lobule structure was disorganized,with the nucleus abnormal at cells’edge,and there were a large number of lipid droplet vacuoles in hepatocytes,while in the experimental group,the hepatic lobular was of clear structure with the number of intracellular lipid droplets significantly decreasing.Compared with group MX,a significant decrease was observed in the body weight and liver wet weight of group E and EC(P<0.01),but only in the liver wet weight of group C(P<0.01).Moreover,the average level of TG,TC,LDL-C,alanine aminotransferase(ALT)and the aspartate aminotransferase(AST),as well as TG and TBARS of group E,C and EC were significantly lower than group MX,while that of HDL-C,GSH-px and SOD was significantly higher than the latter(P<0.01 or P<0.05).There was a synergistic effect between the aerobic exercise and pigment of lycium ruthenicum murr supplement for all the above indicators except AST.Compared with group MX,the gene expression and the protein positive expression of TNFαdecreased and those of PPARαincreased in group E,C and EC(P<0.01 or P<0.05),and the expression of target genes(AOX,CPT1,LCAD,CYP4 A10,CYP4 A12)also increased significantly(P<0.05).There was a synergistic effect between the aerobic exercise and pigment of lycium ruthenicum murr supplement for the gene expression of PPARα,protein positive exression of TNFαand the expression of CPT1,CYP4 A10 mRNA.Conclusions Seven weeks of aerobic exercise and pigment of lycium ruthenicum murr supplement can significantly improve the lipid metabolism disorder in NAFLD mice,which probably promotes liver fatty acid oxidation to reduce lipid accumulation in liver cells,relieving liver lipid degeneration and the damage of liver oxidation,and promoting the benign outcome of NAFLD.
作者
陈伊琳
谢军
陈嘉勤
朱彦
陈锐
屈红林
Chen Yilin;Xie Jun;Chen Jiaqin;Zhu Yan;Chen Rui;Qu Honglin(Key Laboratory of Physical Fitness and Exercise Rehabilitation of Hunan Province,College of Physical Education,Hunan Normal University,Changsha 410012,China;College of Physical Education,Yichun University,Yichun 336000,China)
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
2019年第3期201-210,共10页
Chinese Journal of Sports Medicine
基金
湖南省自然科学资金项目(2016JJ2084)
湖南省研究生创新项目(CX2015B135)
