摘要
目的:从基因启动子区DNA甲基化环节,探讨运动、膳食干预对瘦素抵抗大鼠血清瘦素、下丘脑瘦素受体表达的表观遗传调控机制。方法:130只SD大鼠随机分为空白对照组(n=10)和建模组(n=120),分别给予普通标准啮齿类动物饲料和高脂饲料喂养。后者在第8周时筛选出体重排序前1/3大鼠,并测量其血清瘦素水平,随机分为高脂膳食对照组(HFC)、高脂膳食运动组(HFE)、普通膳食运动组(HRE)、普通膳食对照组(HR)。各组大鼠分别给以中等强度的跑台运动干预或膳食干预8周。喂养周期结束后,禁食,麻醉状态下心脏取血、下丘脑组织。应用重亚硫酸盐修饰直接测序法(bisulfite sequencing-PCR,BSP)检测大鼠血清瘦素基因启动子区(324 bp,-228^+96)25个Cp G位点及下丘脑中瘦素受体基因启动子区(294 bp,-633^-345)20个Cp G位点甲基化变化;同时采用酶联免疫吸附测定血清瘦素含量。结果:1)HFC血清瘦素水平显著高于C(P<0.01);HFE、HRE血清瘦素水平显著低于HFC(P<0.05),且HRE显著低于HFE及HR(P<0.01);2)HFC血清瘦素基因启动子区Cp G位点的平均甲基化程度显著低于C(P<0.01);HFE、HRE血清瘦素基因启动子区Cp G位点的平均甲基化程度显著高于HFC(P<0.05),且HRE血清瘦素基因启动子区Cp G位点的平均甲基化程度显著高于HFE及HR(P<0.01);3)HFC下丘脑瘦素受体基因启动子区Cp G位点的平均甲基化程度显著高于C(P<0.01),HFE、HRE下丘脑瘦素受体基因启动子区Cp G位点的平均甲基化程度显著低于HFC(P<0.01),且HRE下丘脑瘦素受体基因启动子区Cp G位点的平均甲基化程度显著低于HFE及HR(P<0.01)。结论:运动联合膳食干预较单独运动或膳食干预可以明显改善瘦素抵抗,其机制可能是与运动联合膳食干预上调血清瘦素基因启动子区Cp G位点平均甲基化程度、下调下丘脑瘦素受体基因启动子区Cp G位点的平均甲基化程度有关。
objective： From promoter DNA methylation links of the gene,the authors explore the movement,dietary intervention on leptin resistance expression of leptin receptor in hypothalamus of rats serum leptin and epigenetic regulation mechanism. Methods： 130 SD rats were randomly divided into blank control group（ n = 10） and model group（ n = 120）,and were given routine standard rodent animal feed and high fat diet. After 8 weeks,1 /3 of the rats whose weight were heavier in the modeling group were screened out,measured for the serum leptin level and were randomly divided into control group,high fat diet（ HFC）,high fat diet and exercise group（ HFE）,sport in ordinary diet group（ HRE）,normal diet control group（ HR）. The rats were treated with moderate intensity treadmill exercise intervention or dietary intervention for 12 weeks. After the end of the cycle,fasting,anesthesi,heart blood,hypothalamus. Application of sulfite modified direct sequencing（ bisulfite sequencing-PCR,BSP） were detected in plasma leptin gene promoter region（ 324 bp,- 228 - ＋ 96）leptin receptor gene promoter region of 25 CpG loci and the hypothalamus（ 294 bp,- 633 ^- 345） of 20 Cp G locimethylation changes; at the same time by ELISA determination of plasma leptin level adsorption. Results： 1） HFC serum leptin levels were significantly higher than those of C（ P〈 0.01）; the serum leptin levels of HRE and HFE were significantly lower than those of HFC（ P〈0.05）,and HRE were significantly lower than those of HFE and HR（ P〈0.01）. 2）the average methylation degree of HFC in serum leptin gene promoter CpG sites was significantly lower than that of C（ P〈0.01）; the average methylation the degree of HFE,HRE and serum leptin gene promoter CpG sites was significantly higher than that of HFC（ P〈0.05）,the average methylation level of serum leptin and HRE gene promoter CpG sites was significantly higher than that of HFE and HR（ P〈0.01）. 3） the average methylation level of HFC and lept
出处
《沈阳体育学院学报》
CSSCI
北大核心
2016年第3期66-73,共8页
Journal of Shenyang Sport University
基金
教育部人文社会科学研究青年基金项目
编号:13YJC890032
关键词
运动
膳食
干预
瘦素抵抗
瘦素
瘦素受体
exercise
diet
intervention
leptin resistance
leptin
leptin receptor
