本文選題：孕期酒精暴露 + 新生大鼠�。� 參考：《新鄉(xiāng)醫(yī)學(xué)院》2014年碩士論文

【摘要】：背景酗酒是影響人類身體健康的因素之一,不僅有損個人身心健康,對家人和社會都會產(chǎn)生不良影響。尤其是女性孕期飲酒會嚴(yán)重影響胎兒的生長發(fā)育,嚴(yán)重者發(fā)生胎兒酒精綜合征(fetal alcohol syndrome, FAS),中樞神經(jīng)系統(tǒng)失調(diào)便是其特征表現(xiàn)之一。基本節(jié)律性呼吸起源于延髓呼吸中樞,面神經(jīng)后核內(nèi)側(cè)區(qū)是基本節(jié)律性呼吸產(chǎn)生的關(guān)鍵部位。穩(wěn)定的節(jié)律性呼吸是機體存活的前提條件,中樞性呼吸系統(tǒng)疾病,尤其是延髓呼吸中樞部位的病變是臨床工作中的常見病與多發(fā)病,而酗酒引起的子代呼吸中樞功能異常也越來越多見,因此,研究孕期酒精暴露對子代延髓呼吸中樞功能的作用及其機制對相關(guān)疾病的預(yù)防和治療有著廣泛的應(yīng)用前景和重要意義。 目的使用電生理方法研究孕期酒精暴露對子代延髓呼吸中樞基本節(jié)律性呼吸放電(rhythmic respiratory discharge activities, RRDA)、使用Western blot和qRT-PCR檢測α7nACh受體(α7nicotinic acetylcholinergic receptor, α7nAChR)蛋白和基因表達(dá)的變化,探討α7nAChR在孕期酒精暴露中所致子代呼吸抑制中的作用和機制,為臨床預(yù)防和治療提供思路和實驗依據(jù)。 方法Sprague-Dawley大鼠,隨機分為對照組和實驗組。對照組大鼠常規(guī)飼養(yǎng)；實驗組大鼠除常規(guī)飼養(yǎng)外,自合籠前一個月起至哺乳期給予8%的酒精為唯一飲用水。使用2天的新生大鼠,雌雄不拘。 1.記錄反映延髓呼吸中樞功能的腦片RRDA,觀察α7nAChR激動劑乙酰膽堿(acetylcholine, ACh)和拮抗劑銀環(huán)蛇毒素(Alpha-bungarotoxin,α-BGT)對對照組和實驗組RRDA的作用,以明確α7nAChR是否參與孕期酒精暴露對子代延髓呼吸中樞的抑制作用。 2.采用Western Blot技術(shù)檢測α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá),明確孕期酒精暴露是否下調(diào)α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá)。 3.采用qRT-PCR技術(shù)檢測α7nAChR mRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元表達(dá),明確孕期酒精暴露是否下調(diào)α7nAChR mRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元上的表達(dá)。 結(jié)果 1.對照組和實驗組腦片放電60min內(nèi)穩(wěn)定無衰減,實驗?zāi)Ｐ头�(wěn)定可靠。實驗組腦片放電弱于對照組(以對照組為100%,實驗組20min時TI87.67±3.28%、IA90.57±4.81%、RC89.93±4.42%);α7nAChR激動劑ACh對實驗組和對照組腦片放電都有興奮作用,對照組興奮效果強于實驗組(實驗組變化率分別為10.33±0.58%、9.69±0.48%、9.19±0.71%；對照組為：18.33±0.60%、17.55±0.45%、19.56±0.63%);α7nAChR拮抗齊α-BGT對實驗組和對照組腦片放電都有抑制作用,對照組抑制效果強于實驗組(實驗組變化率分別為11.87±0.49%、14.64±0.69%、10.69±0.48%；對照組為：14.64±0.65%、16.89±0.53%、15.56±0.64%); α7nAChR參與孕期酒精暴露對子代延髓呼吸中樞的抑制作用。 2. Western Blot實驗結(jié)果顯示,實驗組α7nAChR在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元表達(dá)水平下調(diào)(對照組1.012±0.090、實驗組0.682±0.034)。 3qRT-PCR實驗結(jié)果顯示,實驗組α7nAChR mRNA在延髓呼吸中樞面神經(jīng)后核內(nèi)側(cè)區(qū)神經(jīng)元表達(dá)水平下調(diào)(對照組1.004±0.101、實驗組0.716±0.047)。 結(jié)論 1.孕期酒精暴露抑制新生大鼠延髓腦片RRDA、抑制延髓呼吸功能。 2.通過下調(diào)α7nAChR和受體mRNA表達(dá)而降低α7nACh對RRRDA的調(diào)節(jié)作用可能是孕期酒精暴露抑制子代延髓中樞呼吸功能的機制之一
[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

【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R714.2

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