孕哺期高蛋白饮食对仔鼠生长、代谢的影响及机制探讨

发布时间：2018-03-30 01:37

  本文选题：高蛋白饮食　切入点：胎盘　出处：《安徽医科大学》2017年硕士论文

【摘要】：目的本研究通过建立孕哺期高蛋白饮食的动物模型,观察孕哺期蛋白质的营养状况对孕期母鼠的生长状况、胎盘的生长发育、胎鼠的宫内生长状况及哺乳期仔鼠的生长、代谢情况的影响,并探讨相关机制,从子代体格发育及成年后慢性病的防治方面提供孕哺期均衡蛋白质摄入的理论依据。方法选取7周龄的SPF级ICR雌性小鼠,适应性喂养一周后受孕。受孕后将孕鼠随机分为两组,即普通饮食的对照组(C)和高蛋白饮食的高蛋白组(H),每组16只孕鼠。母鼠分娩后,每组随机分成两个亚组,即孕期普通饮食哺乳期普通饮食(CC)组、孕期普通饮食哺乳期高蛋白饮食(CH)组、孕期高蛋白饮食哺乳期普通饮食组(HC)和孕期高蛋白饮食哺乳期高蛋白饮食组(HH),每组数量为6窝,每窝10只,喂饲至21天完全断乳。孕期末(GD18),每组取4只孕鼠腹腔注射10%水合氯醛(0.3ml/kg)麻醉处死,分离出胎鼠和胎盘,测量胎鼠体重和胎盘重量,测量血清胆固醇(TC)、甘油三酯(TG)等生化指标。胎盘组织中养分转运相关基因的m RNA表达水平采用RT-PCR的方法来检测,胎盘组织中蛋白的表达水平采用Western Blot的方法来检测;哺乳期间,于分娩后第1天到第18天,每三天测一次母鼠的泌乳量。于仔鼠出生后的第14天(PND14)宰杀一部分,测量各组母鼠的乳汁含量。从仔鼠出生后第一天(PND1)至第21天(PND21),每3天测量仔鼠的体重,于哺乳期末(PND21)腹腔注射10%水合氯醛(0.3ml/kg)麻醉处死,留取血清样本,称量并记录肝脏和肾脏的重量,留取肝肾组织样本。采用罗氏血糖仪测量空腹血糖、试剂盒测量血清TC和TG的水平,采用放射免疫法分析血清胰岛素的水平。结果孕期末,测得高蛋白组胎体比(P0.01)低于对照组,母鼠的胰岛素水平(P0.05),血清总胆固醇水平(P0.05)高于对照组,两组间血清甘油三酯、平均窝重及血糖水平差异均无统计学意义,孕期母鼠体重增长无差异;哺乳期母鼠的泌乳量从第三天起HC组有高于其他三组的趋势,差异无统计学意义。在哺乳期第14天测得的各组间乳汁成分,差异无统计学意义;哺乳期雌、雄仔鼠体重HC组高于其他三组,差异无统计学意义;脏器重量方面:仔鼠出生后第14天(PND14),雄性仔鼠肝脏重量HC、HH组重量均高于CC、CH组(P0.05),雌性仔鼠肝脏重量HC、HH组重量均高于CH组(P0.05);雄性仔鼠肾脏重量HH组重量高于CH组和CC组(P0.05),HC组高于CC组,雌性仔鼠肾脏重量HH组高于CC、CH组;仔鼠出生后第21天(PND21)雄性仔鼠肝脏重量各组间无差异,雌性仔鼠肝脏重量HC组高于CC组(P0.05);雄性仔鼠肾脏重量HC组高于CC组(P0.05),雌性仔鼠肾脏重量HH、HC组均高于CC组;孕期末测得高蛋白组(H)母鼠的血清胰岛素水平显著高于对照组(C)(P0.001),高蛋白组(H)血清TG水平低于对照组(P0.05),血清TC水平差异无统计学意义;高蛋白组(H)胎鼠的血清TG水平高于对照组(P0.05),血糖、血清胰岛素及血清TC水平无差异。在PND14天,CC组仔鼠血清TC水平显著高于HC组和HH组(P0.05);在PND21天,CC组仔鼠血清TC水平显著高于HC组和CH组(P0.05);在PND14天,CC组仔鼠血清TG水平显著高于其他三组(P0.05);在PND21天,CC组和HC组仔鼠血清TG水平显著高于CH组(P0.05)。RT-PCR的结果显示,孕期末,高蛋白组胎盘组织养分转运体CD36(P0.001)和GLUT2(P=0.004)基因的表达水平显著上调;Western-Blot结果显示:高蛋白组胎盘葡糖糖转运体GLUT2蛋白表达水平显著上调(P0.05),差异有统计学意义。结论本次研究结果显示孕哺乳期高蛋白饮食导致小鼠子代血清胰岛素、TG、TC等血清学指标的改变,引起子代生长、代谢的紊乱。RT-PCR和Western-Blot的实验结果显示,孕期高蛋白饮食通过影响胎盘转运率,上调胎盘养分转运体的表达水平,从而影响子代生长、代谢。
[Abstract]:The purpose of this study is through the establishment of animal model of high protein diet during pregnancy and lactation, observation of nutritional status during pregnancy and lactation protein on the growth status of prenatal, placental growth, fetal intrauterine growth and lactation offspring growth, affect metabolism, and discuss the mechanism, provide a theoretical basis for pregnancy and lactation balanced intake of protein from the generation period of physical development and adult chronic disease prevention and control. The SPF level of female ICR mice methods 7 weeks of age, feeding after a week of pregnancy. The pregnant rats were randomly divided into two groups: control group, normal diet (C) high protein group and high protein diet the (H), each group of 16 pregnant rats. Rats in each group after delivery, were randomly divided into two groups, namely normal diet during pregnancy lactation diet (CC) group, normal diet during pregnancy and lactation of high protein diet (CH) group, high protein diet during pregnancy and lactation in general Diet group (HC) and high protein diet during pregnancy and lactation high protein diet group (HH group), the number of 6 nests, 10 per litter, feeding 21 days to complete. At the end of weaning during pregnancy (GD18), each group 4 pregnant rats by intraperitoneal injection of 10% chloral hydrate (0.3ml/kg) anesthesia were isolated fetal and placental weight, placental weight and measurement of fetal rats, serum cholesterol measurement (TC), triglyceride (TG) and other biochemical indicators. The expression level of M RNA in placenta nutrient transporter genes using RT-PCR method to detect the expression level of protein in placenta by Western Blot method to detect lactation; during the first days after birth to eighteenth days, every three days of lactation time were measured. Fourteenth days in rats after birth (PND14) slaughter part, milk content measured female offspring. From the first day (PND1) to twenty-first days (PND21) every 3 days. Measurement of rats weight, feeding on At the end of milk (PND21) intraperitoneal injection of 10% chloral hydrate (0.3ml/kg) anesthesia were collected serum samples, weighing and recording the weight of the liver and kidney, liver and kidney tissue samples collected by Roche. Fasting blood glucose measurement, TC and TG kit to measure serum reagent level, serum insulin was analyzed by radioactive immunoassay the level of the end of pregnancy, the measured high protein group matrix (P0.01) is lower than that of the control group, maternal insulin levels (P0.05), serum total cholesterol (P0.05) was higher than the control group, two groups of serum triglyceride, average weight and blood sugar level differences in nest were not statistically significant, prenatal weight gain no difference; milk yield in lactating rats from the third day HC group was higher than the other three groups the trend, there was no significant difference between the groups. The measured components of milk during lactation for fourteenth days, there was no statistically significant difference; lactating female male rats, weight HC缁勯珮浜庡叾浠栦笁缁，

本文编号：1683802