多巴胺D2受体参与孕期酒精暴露抑制新生大鼠节律性基本呼吸放电
发布时间：2018-12-31 15:31
【摘要】：背景长期过量饮酒对人体造成多器官、复合性严重危害,如心肌病变、造血功能抑制、消化系统溃疡和肿瘤,更为严重的对神经系统和生殖系统造成不可逆的损伤。孕龄妇女摄入酒精除对自身产生损害外还会对腹中胎儿产生严重、不可挽回的损伤。研究发现孕期摄入酒精降低子代低氧条件下摄取氧的能力；大鼠孕期摄入酒精降低新生大鼠延髓离体脑片呼吸节律性放电。这样的实验结果发现孕期酒精暴露抑制由各级呼吸中枢的共同参与调节的动物整体水平上的呼吸功能,产生基本呼吸节律性放电的延髓呼吸中枢也受到了抑制。基本节律性呼吸起源于延髓呼吸中枢,前包钦格复合体(pre-Botzinger complex, pre-BotC)位于延髓腹外侧区,是延髓基本节律性呼吸产生的关键部位。多巴胺是哺乳动物体内广泛存在的儿茶酚胺类神经递质,调节着多种生理活动,在中枢神经系统内多巴胺通过多巴胺受体参与情绪、运动、摄食、饥饿、内分泌等生理活动。现已发现延髓神经元细胞膜上存在多巴胺D2受体,但该受体是否参与孕期摄入酒精对新生子代延髓呼吸中枢的抑制作用的研究尚未见报道,因此我们设计本实验,利用神经电生理和分子生物学的实验方法研究多巴胺受体2是否参与该抑制作用及其可能机制。目的研究孕期酒精暴露对新生大鼠延髓呼吸中枢节律性基本呼吸放电(rhythmic respiratory discharge activities, RRDA)的作用和多巴胺D2受体在其中的作用。方法对成年健康Sprague-Dawley大鼠进行孕期酒精暴露,自合笼前30d至实验时饲喂大鼠自主饮用且只饮用8%酒精水溶液；对照大鼠常规饲养。本实验使两组用2d新生大鼠,雌雄不拘。1.RRDA记录：制作2d新生大鼠延髓离体脑片,使用吸附电极在脑片腹侧面吸附舌下神经根,舌下神经根传出的电活动即为反映了延髓呼吸功能的节律性基本呼吸放电。在2组新生大鼠脑片灌流液中加入多巴胺受体2特异性激动剂Quinpirole和特异性拮抗剂Raclopride,观察Quinpirole和IRaclopride对两组RRDA的作用差异,以明确孕期酒精暴露是否改变多巴胺2受体对新生大鼠延髓离体脑片节律性基本呼吸放电作用效果。2.将2d新生小鼠延髓离体脑片pre-BotC区切下,采用Western Blot检验多巴胺D2受体在pre-BotC区神经元上的表达,研究孕期酒精暴露对受体表达的作用。3.将2d新生小鼠延髓离体脑片pre-BotC区切下,采用qRT-PCR-技术检测多巴胺D2受体mRNA在延髓呼吸中枢pre-BotC区神经元上的表达,研究孕期酒精暴露对受体mRNA表达的作用。结果1.对照组和酒精暴露组新生大鼠延髓离体脑片呼吸节律性放电在60min内各时间点之间无统计学改变,实验模型成立。多巴胺D2受体激动剂Quinpirole抑制对照组和酒精暴露组脑片呼吸节律性放电,对对照组的抑制效果强于实验组；多巴胺D2受体拮抗剂Raclopride对两组新生大鼠延髓离体脑片呼吸节律性放电都有兴奋作用,酒精暴露组放电变化程度低于对照组。2. Western Blot实验发现,孕期酒精暴露降低新生大鼠延髓呼吸中枢pre-BotC区神经元多巴胺D2受体表达水平。3. qRT-PCR实验显示,与对照组相比,孕期酒精暴露降低新生大鼠延髓呼吸中枢pre-BotC区神经元多巴胺D2受体mRNA表达水平。结论1.孕期酒精暴露抑制新生大鼠延髓脑片节律性基本呼吸放电,降低多巴胺D2受体对脑片节律性基本呼吸放电的调节作用。2.孕期酒精暴露下调延髓前包钦格复合体神经元多巴胺D2受体水平和受体mRNA表达水平,这可能是多巴胺D2受体参与孕期酒精暴露抑制子代延髓脑片律性基本呼吸放电机制之一。
[Abstract]:Background: Long-term excessive drinking can cause multiple organs and compound serious harm to the human body, such as cardiomyopathy, hemopoietic function inhibition, digestive system ulceration and tumor, and more serious irreversible damage to the nervous system and the reproductive system. The intake of alcohol in pregnant women may also cause serious and irreversible damage to the fetus in the abdomen, in addition to its own damage. The study found that the intake of alcohol during pregnancy decreased the ability of oxygen to be taken under the low-oxygen condition of the offspring, and the intake of alcohol during pregnancy decreased the respiratory rhythm of the medulla oblongata of the neonatal rats. Such experimental results showed that during pregnancy, alcohol exposure inhibited the respiratory function at the whole level of the animal regulated by the co-participation of the respiratory center at all levels, and the medulla oblongata respiratory center, which produced the basic respiratory rhythm discharge, was also inhibited. The basic rhythmic breathing originates from the medulla oblongata respiratory center, and the pre-Botzinger complex (pre-BopoC) is located in the lateral medulla of the medulla oblongata, which is the key part of the basic rhythm of the medulla oblongata. Dopamine is a widely available catecholamine neurotransmitter in a mammal, which regulates a variety of physiological activities in which dopamine is involved in physiological activities such as mood, exercise, food consumption, hunger, and endocrine. It has been found that there is a dopamine D2 receptor on the cell membrane of the medulla oblongata, but the study of whether the receptor is involved in the inhibition of the uptake of alcohol during pregnancy on the respiratory center of the medulla oblongata is not reported, so we design this experiment, Whether the dopamine receptor 2 is involved in the inhibition and its possible mechanism is studied by means of the experimental methods of neuroelectrophysiology and molecular biology. Objective To study the effects of alcohol exposure during pregnancy on the basic respiratory discharge (RRDA) and the role of dopamine D2 receptor in neonatal rats. Methods The adult healthy Sprague-Dawley rats were exposed to alcohol during pregnancy. In this experiment, two groups of newborn rats were divided into two groups. The first. RRDA records: to make 2d neonatal rat's medulla oblongata, and to use the adsorption electrode to adsorb the hypoglossal nerve roots on the ventral side of the brain, and the electrical activity from the root of the hypoglossal nerve is the basic respiratory discharge that reflects the respiratory function of the medulla oblongata. The effect of Quinpirole and IRaclopride on the two groups of RRDA was observed by the addition of a dopamine receptor 2-specific agonist, Quinpirole, and a specific antagonist, Racopride, to the brain slices of the 2-group neonatal rats. in order to determine whether the exposure of the alcohol to the dopamine-2 receptor changes the effect of the dopamine-2 receptor on the basic respiratory discharge of the medulla oblongata in the newborn rats. The expression of the dopamine D2 receptor in the pre-BopoC region was examined by Western Blot, and the effect of alcohol exposure on the expression of the receptor was studied. The expression of dopamine D2 receptor mRNA in the pre-BopoC region of the medulla oblongata was detected by the qRT-PCR-technique, and the effect of alcohol exposure on the expression of the receptor mRNA was studied. Results 1. In the control group and the alcohol exposure group, the respiratory rhythm of the medulla oblongata was not statistically changed between the time points within 60min, and the experimental model was established. in that control group, the dopamine D2 receptor agonist, Quinn pirole, inhibited the respiratory rhythm of the brain slices of the control group and the alcohol exposure group, and the inhibition effect on the control group was stronger than that of the experimental group; the dopamine D2 receptor antagonist Racopride has an excitation effect on the respiratory rhythm of the medulla oblongata of the two groups of newborn rats, The degree of discharge of the alcohol exposure group was lower than that of the control group. Western Blot experiment found that alcohol exposure during pregnancy decreased the level of dopamine D2 receptor expression in the pre-BopoC region of the neonatal rat's medulla oblongata. The qRT-PCR showed that the level of the expression of dopamine D2 receptor mRNA in the pre-BopoC region of the medulla oblongata of the neonatal rats was reduced by alcohol exposure during pregnancy as compared to the control group. Conclusion 1. The effect of the dopamine D2 receptor on the basic respiratory discharge of the brain slices was reduced by the basic respiratory discharge in the brain of the neonatal rat by alcohol exposure during pregnancy. The level of the dopamine D2 receptor and the expression of the receptor mRNA in the neurons of the submedulla oblongata complex are reduced by alcohol exposure during pregnancy, which may be one of the basic respiratory discharge mechanisms of the dopamine D2 receptor involved in the inhibition of alcohol exposure during pregnancy.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R714
本文编号：2396773
[Abstract]:Background: Long-term excessive drinking can cause multiple organs and compound serious harm to the human body, such as cardiomyopathy, hemopoietic function inhibition, digestive system ulceration and tumor, and more serious irreversible damage to the nervous system and the reproductive system. The intake of alcohol in pregnant women may also cause serious and irreversible damage to the fetus in the abdomen, in addition to its own damage. The study found that the intake of alcohol during pregnancy decreased the ability of oxygen to be taken under the low-oxygen condition of the offspring, and the intake of alcohol during pregnancy decreased the respiratory rhythm of the medulla oblongata of the neonatal rats. Such experimental results showed that during pregnancy, alcohol exposure inhibited the respiratory function at the whole level of the animal regulated by the co-participation of the respiratory center at all levels, and the medulla oblongata respiratory center, which produced the basic respiratory rhythm discharge, was also inhibited. The basic rhythmic breathing originates from the medulla oblongata respiratory center, and the pre-Botzinger complex (pre-BopoC) is located in the lateral medulla of the medulla oblongata, which is the key part of the basic rhythm of the medulla oblongata. Dopamine is a widely available catecholamine neurotransmitter in a mammal, which regulates a variety of physiological activities in which dopamine is involved in physiological activities such as mood, exercise, food consumption, hunger, and endocrine. It has been found that there is a dopamine D2 receptor on the cell membrane of the medulla oblongata, but the study of whether the receptor is involved in the inhibition of the uptake of alcohol during pregnancy on the respiratory center of the medulla oblongata is not reported, so we design this experiment, Whether the dopamine receptor 2 is involved in the inhibition and its possible mechanism is studied by means of the experimental methods of neuroelectrophysiology and molecular biology. Objective To study the effects of alcohol exposure during pregnancy on the basic respiratory discharge (RRDA) and the role of dopamine D2 receptor in neonatal rats. Methods The adult healthy Sprague-Dawley rats were exposed to alcohol during pregnancy. In this experiment, two groups of newborn rats were divided into two groups. The first. RRDA records: to make 2d neonatal rat's medulla oblongata, and to use the adsorption electrode to adsorb the hypoglossal nerve roots on the ventral side of the brain, and the electrical activity from the root of the hypoglossal nerve is the basic respiratory discharge that reflects the respiratory function of the medulla oblongata. The effect of Quinpirole and IRaclopride on the two groups of RRDA was observed by the addition of a dopamine receptor 2-specific agonist, Quinpirole, and a specific antagonist, Racopride, to the brain slices of the 2-group neonatal rats. in order to determine whether the exposure of the alcohol to the dopamine-2 receptor changes the effect of the dopamine-2 receptor on the basic respiratory discharge of the medulla oblongata in the newborn rats. The expression of the dopamine D2 receptor in the pre-BopoC region was examined by Western Blot, and the effect of alcohol exposure on the expression of the receptor was studied. The expression of dopamine D2 receptor mRNA in the pre-BopoC region of the medulla oblongata was detected by the qRT-PCR-technique, and the effect of alcohol exposure on the expression of the receptor mRNA was studied. Results 1. In the control group and the alcohol exposure group, the respiratory rhythm of the medulla oblongata was not statistically changed between the time points within 60min, and the experimental model was established. in that control group, the dopamine D2 receptor agonist, Quinn pirole, inhibited the respiratory rhythm of the brain slices of the control group and the alcohol exposure group, and the inhibition effect on the control group was stronger than that of the experimental group; the dopamine D2 receptor antagonist Racopride has an excitation effect on the respiratory rhythm of the medulla oblongata of the two groups of newborn rats, The degree of discharge of the alcohol exposure group was lower than that of the control group. Western Blot experiment found that alcohol exposure during pregnancy decreased the level of dopamine D2 receptor expression in the pre-BopoC region of the neonatal rat's medulla oblongata. The qRT-PCR showed that the level of the expression of dopamine D2 receptor mRNA in the pre-BopoC region of the medulla oblongata of the neonatal rats was reduced by alcohol exposure during pregnancy as compared to the control group. Conclusion 1. The effect of the dopamine D2 receptor on the basic respiratory discharge of the brain slices was reduced by the basic respiratory discharge in the brain of the neonatal rat by alcohol exposure during pregnancy. The level of the dopamine D2 receptor and the expression of the receptor mRNA in the neurons of the submedulla oblongata complex are reduced by alcohol exposure during pregnancy, which may be one of the basic respiratory discharge mechanisms of the dopamine D2 receptor involved in the inhibition of alcohol exposure during pregnancy.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R714
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