本文選題：高蛋白飲食　切入點：胎盤　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的本研究通過建立孕哺期高蛋白飲食的動物模型,觀察孕哺期蛋白質(zhì)的營養(yǎng)狀況對孕期母鼠的生長狀況、胎盤的生長發(fā)育、胎鼠的宮內(nèi)生長狀況及哺乳期仔鼠的生長、代謝情況的影響,并探討相關(guān)機(jī)制,從子代體格發(fā)育及成年后慢性病的防治方面提供孕哺期均衡蛋白質(zhì)攝入的理論依據(jù)。方法選取7周齡的SPF級ICR雌性小鼠,適應(yīng)性喂養(yǎng)一周后受孕。受孕后將孕鼠隨機(jī)分為兩組,即普通飲食的對照組(C)和高蛋白飲食的高蛋白組(H),每組16只孕鼠。母鼠分娩后,每組隨機(jī)分成兩個亞組,即孕期普通飲食哺乳期普通飲食(CC)組、孕期普通飲食哺乳期高蛋白飲食(CH)組、孕期高蛋白飲食哺乳期普通飲食組(HC)和孕期高蛋白飲食哺乳期高蛋白飲食組(HH),每組數(shù)量為6窩,每窩10只,喂飼至21天完全斷乳。孕期末(GD18),每組取4只孕鼠腹腔注射10%水合氯醛(0.3ml/kg)麻醉處死,分離出胎鼠和胎盤,測量胎鼠體重和胎盤重量,測量血清膽固醇(TC)、甘油三酯(TG)等生化指標(biāo)。胎盤組織中養(yǎng)分轉(zhuǎn)運相關(guān)基因的m RNA表達(dá)水平采用RT-PCR的方法來檢測,胎盤組織中蛋白的表達(dá)水平采用Western Blot的方法來檢測;哺乳期間,于分娩后第1天到第18天,每三天測一次母鼠的泌乳量。于仔鼠出生后的第14天(PND14)宰殺一部分,測量各組母鼠的乳汁含量。從仔鼠出生后第一天(PND1)至第21天(PND21),每3天測量仔鼠的體重,于哺乳期末(PND21)腹腔注射10%水合氯醛(0.3ml/kg)麻醉處死,留取血清樣本,稱量并記錄肝臟和腎臟的重量,留取肝腎組織樣本。采用羅氏血糖儀測量空腹血糖、試劑盒測量血清TC和TG的水平,采用放射免疫法分析血清胰島素的水平。結(jié)果孕期末,測得高蛋白組胎體比(P0.01)低于對照組,母鼠的胰島素水平(P0.05),血清總膽固醇水平(P0.05)高于對照組,兩組間血清甘油三酯、平均窩重及血糖水平差異均無統(tǒng)計學(xué)意義,孕期母鼠體重增長無差異;哺乳期母鼠的泌乳量從第三天起HC組有高于其他三組的趨勢,差異無統(tǒng)計學(xué)意義。在哺乳期第14天測得的各組間乳汁成分,差異無統(tǒng)計學(xué)意義;哺乳期雌、雄仔鼠體重HC組高于其他三組,差異無統(tǒng)計學(xué)意義;臟器重量方面:仔鼠出生后第14天(PND14),雄性仔鼠肝臟重量HC、HH組重量均高于CC、CH組(P0.05),雌性仔鼠肝臟重量HC、HH組重量均高于CH組(P0.05);雄性仔鼠腎臟重量HH組重量高于CH組和CC組(P0.05),HC組高于CC組,雌性仔鼠腎臟重量HH組高于CC、CH組;仔鼠出生后第21天(PND21)雄性仔鼠肝臟重量各組間無差異,雌性仔鼠肝臟重量HC組高于CC組(P0.05);雄性仔鼠腎臟重量HC組高于CC組(P0.05),雌性仔鼠腎臟重量HH、HC組均高于CC組;孕期末測得高蛋白組(H)母鼠的血清胰島素水平顯著高于對照組(C)(P0.001),高蛋白組(H)血清TG水平低于對照組(P0.05),血清TC水平差異無統(tǒng)計學(xué)意義;高蛋白組(H)胎鼠的血清TG水平高于對照組(P0.05),血糖、血清胰島素及血清TC水平無差異。在PND14天,CC組仔鼠血清TC水平顯著高于HC組和HH組(P0.05);在PND21天,CC組仔鼠血清TC水平顯著高于HC組和CH組(P0.05);在PND14天,CC組仔鼠血清TG水平顯著高于其他三組(P0.05);在PND21天,CC組和HC組仔鼠血清TG水平顯著高于CH組(P0.05)。RT-PCR的結(jié)果顯示,孕期末,高蛋白組胎盤組織養(yǎng)分轉(zhuǎn)運體CD36(P0.001)和GLUT2(P=0.004)基因的表達(dá)水平顯著上調(diào);Western-Blot結(jié)果顯示:高蛋白組胎盤葡糖糖轉(zhuǎn)運體GLUT2蛋白表達(dá)水平顯著上調(diào)(P0.05),差異有統(tǒng)計學(xué)意義。結(jié)論本次研究結(jié)果顯示孕哺乳期高蛋白飲食導(dǎo)致小鼠子代血清胰島素、TG、TC等血清學(xué)指標(biāo)的改變,引起子代生長、代謝的紊亂。RT-PCR和Western-Blot的實驗結(jié)果顯示,孕期高蛋白飲食通過影響胎盤轉(zhuǎn)運率,上調(diào)胎盤養(yǎng)分轉(zhuǎn)運體的表達(dá)水平,從而影響子代生長、代謝。
[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緇勯珮浜庡叾浠栦笁緇，

本文編號：1683802