a7nACh受体在孕期酒精暴露抑制新生大鼠RRDA中的作用

发布时间：2018-05-11 14:10

本文选题：孕期酒精暴露 + 新生大鼠　；参考：《新乡医学院》2014年硕士论文

【摘要】：背景酗酒是影响人类身体健康的因素之一,不仅有损个人身心健康,对家人和社会都会产生不良影响。尤其是女性孕期饮酒会严重影响胎儿的生长发育,严重者发生胎儿酒精综合征(fetal alcohol syndrome, FAS),中枢神经系统失调便是其特征表现之一。基本节律性呼吸起源于延髓呼吸中枢,面神经后核内侧区是基本节律性呼吸产生的关键部位。稳定的节律性呼吸是机体存活的前提条件,中枢性呼吸系统疾病,尤其是延髓呼吸中枢部位的病变是临床工作中的常见病与多发病,而酗酒引起的子代呼吸中枢功能异常也越来越多见,因此,研究孕期酒精暴露对子代延髓呼吸中枢功能的作用及其机制对相关疾病的预防和治疗有着广泛的应用前景和重要意义。目的使用电生理方法研究孕期酒精暴露对子代延髓呼吸中枢基本节律性呼吸放电(rhythmic respiratory discharge activities, RRDA)、使用Western blot和qRT-PCR检测α7nACh受体(α7nicotinic acetylcholinergic receptor, α7nAChR)蛋白和基因表达的变化,探讨α7nAChR在孕期酒精暴露中所致子代呼吸抑制中的作用和机制,为临床预防和治疗提供思路和实验依据。方法Sprague-Dawley大鼠,随机分为对照组和实验组。对照组大鼠常规饲养；实验组大鼠除常规饲养外,自合笼前一个月起至哺乳期给予8%的酒精为唯一饮用水。使用2天的新生大鼠,雌雄不拘。 1.记录反映延髓呼吸中枢功能的脑片RRDA,观察α7nAChR激动剂乙酰胆碱(acetylcholine, ACh)和拮抗剂银环蛇毒素(Alpha-bungarotoxin,α-BGT)对对照组和实验组RRDA的作用,以明确α7nAChR是否参与孕期酒精暴露对子代延髓呼吸中枢的抑制作用。 2.采用Western Blot技术检测α7nAChR在延髓呼吸中枢面神经后核内侧区神经元上的表达,明确孕期酒精暴露是否下调α7nAChR在延髓呼吸中枢面神经后核内侧区神经元上的表达。 3.采用qRT-PCR技术检测α7nAChR mRNA在延髓呼吸中枢面神经后核内侧区神经元表达,明确孕期酒精暴露是否下调α7nAChR mRNA在延髓呼吸中枢面神经后核内侧区神经元上的表达。结果 1.对照组和实验组脑片放电60min内稳定无衰减,实验模型稳定可靠。实验组脑片放电弱于对照组(以对照组为100%,实验组20min时TI87.67±3.28%、IA90.57±4.81%、RC89.93±4.42%);α7nAChR激动剂ACh对实验组和对照组脑片放电都有兴奋作用,对照组兴奋效果强于实验组(实验组变化率分别为10.33±0.58%、9.69±0.48%、9.19±0.71%；对照组为：18.33±0.60%、17.55±0.45%、19.56±0.63%);α7nAChR拮抗齐α-BGT对实验组和对照组脑片放电都有抑制作用,对照组抑制效果强于实验组(实验组变化率分别为11.87±0.49%、14.64±0.69%、10.69±0.48%；对照组为：14.64±0.65%、16.89±0.53%、15.56±0.64%); α7nAChR参与孕期酒精暴露对子代延髓呼吸中枢的抑制作用。 2. Western Blot实验结果显示,实验组α7nAChR在延髓呼吸中枢面神经后核内侧区神经元表达水平下调(对照组1.012±0.090、实验组0.682±0.034)。 3qRT-PCR实验结果显示,实验组α7nAChR mRNA在延髓呼吸中枢面神经后核内侧区神经元表达水平下调(对照组1.004±0.101、实验组0.716±0.047)。结论 1.孕期酒精暴露抑制新生大鼠延髓脑片RRDA、抑制延髓呼吸功能。 2.通过下调α7nAChR和受体mRNA表达而降低α7nACh对RRRDA的调节作用可能是孕期酒精暴露抑制子代延髓中枢呼吸功能的机制之一
[Abstract]:Drinking in the background is one of the factors that affect the health of the human body. It not only damages the physical and mental health of the individual, but also has a bad effect on the family and society. In particular, drinking in women during pregnancy will seriously affect the growth and development of the fetus, and the severe fetal alcohol syndrome (FAS) is the main characteristic of the central nervous system. One of the manifestations. Basic rhythmic respiration originates in the medullary respiratory center. The medial region of the posterior nucleus of the facial nerve is the key part of the basic rhythmic respiration. Stable rhythmic respiration is the precondition for the survival of the body. Central respiratory diseases, especially the lesions of the medulla of the medulla, are common and frequently occurring diseases in the clinical work. The dysfunction of the respiratory central function of the offspring is also more and more common. Therefore, the study of the effect and mechanism of alcohol exposure on the medullary respiratory central function during pregnancy and its mechanism have broad application prospects and important significance for the prevention and treatment of related diseases.
Objective to study the basic rhythmic respiratory discharge (rhythmic respiratory discharge activities, RRDA) in the medullary respiratory center of the offspring during pregnancy by electrophysiological method, and to detect the changes of alpha 7nACh receptor (alpha 7nicotinic acetylcholinergic receptor, alpha) protein and gene expression by Western blot and qRT-PCR. The role and mechanism of prenatal alcohol exposure in the respiratory suppression of offspring is to provide ideas and experimental evidence for clinical prevention and treatment.
Methods Sprague-Dawley rats were randomly divided into the control group and the experimental group. The rats in the control group were reared routinely. The rats in the experimental group were given 8% alcohol as the only drinking water from the first month of the cage to the lactation period except the conventional feeding. The female and male rats were used for 2 days.
1. the brain slice RRDA reflecting the function of the medulla's respiratory center was recorded. The effects of alpha 7nAChR agonist (acetylcholine, ACh) and antagonist, Alpha-bungarotoxin, alpha -BGT, on RRDA in the control group and the experimental group were observed to determine whether alpha 7nAChR was involved in the inhibitory effect of alcohol sperm exposure on the medullary respiratory center of the offspring.
2. Western Blot technique was used to detect the expression of alpha 7nAChR on the neurons in the medial nucleus of the medulla of the medulla of the medulla of the medulla. It was clear whether the alcohol exposure during pregnancy reduced the expression of alpha 7nAChR on the neurons in the medial nucleus of the medulla of the medulla of the medulla.
3. qRT-PCR technique was used to detect the expression of alpha 7nAChR mRNA in the medial nucleus of the medulla of the medulla of the medulla and the expression of the neurons in the medial region of the posterior nucleus of the medulla of the medulla. It was clear whether the alcohol exposure during pregnancy reduced the expression of alpha 7nAChR mRNA on the neurons in the medial nucleus of the medulla of the medulla of the medulla.
Result
The brain slices of 1. control group and experimental group were stable and unattenuated, and the experimental model was stable and reliable. The discharge of brain slices in the experimental group was weaker than that of the control group (100% in the control group, 100% in the control group, TI87.67 + 3.28%, IA90.57 + 4.81% and RC89.93 4.42%) in the experimental group, and the alpha 7nAChR agonist ACh had the excitatory effect on the discharge of the brain slices in the experimental group and the control group, and the control group was excited by the excitation effect. The results were stronger than that in the experimental group (10.33 + 0.58%, 9.69 + 0.48%, 9.19 + 0.71%, respectively, 18.33 + 0.60%, 17.55 + 0.45%, 19.56 + 0.63%), and alpha 7nAChR antagonistic Qi -BGT had inhibitory effect on the discharge of brain slices in the experimental group and the control group, and the control group was stronger than the experimental group (the experimental group was 11.87 + 0.49%, respectively). It was 4 + 0.69%, 10.69 + 0.48%, and the control group was 14.64 + 0.65%, 16.89 + 0.53%, 15.56 + 0.64%, and alpha 7nAChR was involved in the inhibitory effect of alcohol exposure on the medullary respiratory center of the offspring.
The results of 2. Western Blot test showed that the expression level of alpha 7nAChR in the medial nucleus of the medulla of the medulla of the medulla was down (1.012 + 0.090 in the control group and 0.682 + 0.034 in the experimental group).
The results of 3qRT-PCR test showed that the expression level of alpha 7nAChR mRNA in the medial nucleus of the medulla of the medulla of the medulla was down (1.004 + 0.101 in the control group and 0.716 + 0.047 in the experimental group).
conclusion
1. alcohol exposure during pregnancy inhibited RRDA in the medullary slices of neonatal rats and inhibited medulla oblongata respiratory function.
2. reducing the regulation of alpha 7nACh to RRRDA by reducing the expression of alpha 7nAChR and receptor mRNA may be one of the mechanisms of the central respiratory function of the medulla oblongata in the pregnancy induced by alcohol exposure

【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R714.2

【参考文献】