宫内发育迟缓大鼠脑组织中生长相关蛋白-43表达的研究

发布时间：2018-04-11 01:00

  本文选题：宫内发育迟缓 + 生长相关蛋白-43　； 参考：《昆明医科大学》2012年硕士论文

【摘要】：目的：本研究通过孕期全程低蛋白饮食法建立宫内发育迟缓(intrauterine growth retardation, IUGR)的实验动物模型,观察生长相关蛋白-43(growth associated protein-43,GAP-43)在IUGR大鼠脑组织中不同部位及不同发育阶段的表达变化情况,分析GAP-43在IUGR大鼠发生脑发育及功能异常中的作用机制,为临床干预及治疗提供相关理论依据。 方法：①采用孕期全程低蛋白饮食法建立IUGR实验动物模型。②母鼠按受孕顺序随机分为低蛋白饮食组和正常饮食组。低蛋白饮食组中,符合IUGR诊断标准的仔鼠按随机原则抽取30只作为IUGR组；正常饮食组中,正常出生体重仔鼠按随机原则抽取30只作为对照组；分别将两组实验仔鼠按随机原则分为出生后0天、10天、30天三个亚组。③两组实验仔鼠分别在出生后0天、10天、30天计量体重及脑重。④用HE染色法观察两组实验仔鼠在不同发育阶段脑组织的病理特征。⑤用免疫组化法观察两组实验仔鼠在不同生长发育阶段脑皮质内及海马CA1区、CA3区GAP-43蛋白的表达改变情况。⑥荧光定量聚合酶链反应(fluorescence quantitative polymerase chain reaction,FQ-PCR)法检测两组实验仔鼠在不同生长发育阶段大脑海马区GAP-43mRNA的表达改变情况。 结果：①低蛋白饮食组IUGR发生率(62.5%)显著高于正常饮食组(7.89%,P0.05)。②低蛋白饮食组死胎率(17.94%)显著高于正常饮食组(5.0%,P0.05)。③IUGR组仔鼠出生后0天体重(4.6847±0.3126g)显著低于对照组(6.4352±0.3135g,P0.01)；出生后10天、30天两组体重无明显差异(P0.05)。④两组仔鼠脑重在出生后0天、10天、30天时,IUGR组均明显低于对照组,差异有统计学意义(P0.01)。⑤GAP-43蛋白在仔鼠大脑皮质区的表达,IUGR组出生后0天、10天均明显低于对照组(P0.01)；出生后30天无明显差异(P0.05)。⑥GAP-43蛋白在仔鼠海马CA1区、CA3区的表达,出生后10天IUGR组明显低于对照组(P0.01),出生后0天、30天无明显差异(P0.05)。⑦FQ-PCR相对定量结果示两组仔鼠出生0天时海马区GAP-43mRNA的表达水平无明显差异(P0.05)；出生后10天IUGR组明显低于对照组,差异有统计学意义(P0.01),出生后30天无明显差异(P0.05)。 结论： 1.孕期全程低蛋白饮食法为较理想的IUGR造模方法。 2. IUGR组仔鼠脑质量的发育在较长时期内低于对照组。 3. IUGR组仔鼠出生时即存在脑皮质发育异常,并持续至出生后30天(约人类8岁)。GAP-43在海马区的低表达可能与IUGR儿发生远期学习、记忆等智能发育障碍有关。
[Abstract]:Objective: To study the method of whole pregnancy low protein diet to establish intrauterine growth retardation (intrauterine growth, retardation, IUGR) of the experimental animal model, to observe the growth associated protein -43 (growth associated protein-43, GAP-43) in the brain tissue of IUGR rats and the expression of different parts in different developmental stages, GAP-43 analysis of mechanism of brain development and abnormal function in IUGR rats, to provide theoretical basis for clinical treatment and intervention.
Method: to establish the animal model of IUGR by the method of whole pregnancy low protein diet. The rats in pregnancy were randomly divided into the low protein diet group and normal diet group. The low protein diet group, according to the diagnosis criteria of IUGR rats were randomly selected 30 rats as IUGR group; normal diet group, normal birth weight rats were randomly selected 30 rats as control group; the two groups of experimental rats were randomly divided into 0 days, 10 days after birth, 30 days and three sub groups. The two groups of experimental rats were born after 0 days, 10 days, 30 days to measure the weight of body and brain. Using HE staining to observe the pathological features of two groups of experimental rats at different developmental stages of brain tissue. The immunohistochemistry was used to observe two groups of experimental rats at different developmental stages in the cerebral cortex and hippocampal CA1 region, the expression of CA3 GAP-43 protein. The changes of fluorescence quantitative polymerase chain Fluorescence (quantitative polymerase chain reaction, FQ-PCR) method was used to detect the expression of GAP-43mRNA in the hippocampus of two groups of experimental offspring at different growth stages.
缁撴灉锛氣憼浣庤泲鐧介ギ椋熺粍IUGR鍙戠敓鐜，

本文编号：1733699