哺乳期使用SSRI对子代大鼠行为学及5-羟色胺能神经元环路功能影响的研究

发布时间：2018-04-11 09:26

本文选题：哺乳期 + SSRI　；参考：《昆明医科大学》2016年硕士论文

【摘要】：[目的]本实验通过建立哺乳期使用氟西汀大鼠动物模型,模拟发育期高血5-羟色胺自闭症动物模型,观察哺乳期使用氟西汀对子代大鼠行为学、5-羟色胺能神经元环路功能的影响,从而探讨哺乳期使用SSRI类抗抑郁药对子代行为学影响及其可能的病理生理机制,以期为自闭症发病机制的研究提供理论依据。[方法]将SD妊娠大鼠随机分成2组,每组各3只。待分娩后第5-21d予以实验组大鼠氟西汀(12mg/kg),对照组大鼠予以等量的生理盐水,期间正常哺乳。婴儿期分别测量子代大鼠体重、毛发、睁眼、听觉和自由悬挂实验情况。待子代出生后第21天断奶并将雌雄大鼠分笼饲养。儿童期子代大鼠行埋藏食物小球实验和旷场试验。儿童期(P35d)和青年期(P75d)分别取材行Western-Blot、IHC、IF检测子代大鼠前额皮质和中缝核SERT及TPH的含量。同时,应用ELISA技术检测儿童期和青年期前额皮质和中缝核5-HT含量。[结果]一、行为学实验结果：1.体重：与对照组相比,氟西汀组体重9d,11d,13d均较对照组明显降低,有显著性差异(P0.05)；2.毛发：与对照组相比,氟西汀组毛发长度9d,11d无明显降低,没有显著性差异(P0.05)。在13d实验组与对照组相比明显降低,有显著性差异(P0.05)：3.睁眼：13d氟西汀组与对照组相比睁眼程度无明显改变(P0.05)；4.听觉：与对照组相比,氟西汀组听觉敏感性9d,11d明显增加,有显著性差异(P0.01)。13d实验组与对照组相比听觉敏感性无明显增加,无显著性差异(P0.05)；5.埋藏食物小球实验：与对照组相比,氟西汀组仔鼠埋藏食物小球实验时间26d无明显改变,无显著性差异(P0.05)；6.自由悬挂实验：与对照组相比,氟西汀组自由悬挂实验时间在1ld,13d均明显减少,有显著性差异(P0.01)；7.旷场实验：与对照组相比,氟西汀组大便粒数在28d,29d,30d,31d均明显增加,有显著性差异(P0.01)；氟西汀组大便粒数平均值也有显著性差异(P0.01)。二、免疫印迹、免疫组化、免疫荧光法检测前额皮质和中缝核SERT和TPH蛋白；酶联免疫吸附法检测前额叶皮质和中缝核5-HT的含量：1.应用Western-blot、IHC、IF技术检测儿童期(35d)和青年期(75d) SERT蛋白在前额皮质和中缝核表达,结果发现：与哺乳期盐水暴露组相比,FLX暴露组儿童期和青年期SERT蛋白表达明显下调,有显著性差异(P0.05)。2.应用Western-blot、IHC、IF技术检测儿童期(35d)和青年期(75d)TPH蛋白在前额皮质和中缝核表达,结果发现：与哺乳期盐水暴露组子代相比,FLX暴露组儿童TPH蛋白表达明显下调,有显著性差异(P0.05)。3.应用IF技术检测仔鼠前额皮质神经元轴突及数目,结果发现：氟西汀暴露组仔鼠儿童期(35d)和青年期(75d)前额叶皮质神经元数量和轴突明显减少。4.酶联免疫吸附法检测5-HT在前额叶皮质和中缝核含量,结果发现：与对照组相比,儿童期(35d)氟西汀组5-HT在大脑总含量明显增高,有显著性差异(P0.01)；婴儿期(75d)氟西汀组5-HT在前额皮质和中缝核含量均明显增高,有显著性差异(P0.05)。[结论]1.哺乳期暴露于氟西汀导致子代的发育和行为异常。2.哺乳期暴露于氟西汀可通过下调子代大鼠前额皮质和中缝核中TPH、SERT蛋白表达提高5-HT含量。3.暴露于氟西汀子代脑组织5-HT含量升高,可能导致类自闭行为。4.哺乳期暴露于SSRIs类抗抑郁药对子代大鼠行为学有显著影响,这种影响可能是由5-羟色胺能神经元系统紊乱所致。
[Abstract]:[Objective] to establish lactation rats fluoxetine animal models used by this experiment, simulated development stage of high blood 5- HT autism animal model, to observe the lactation of fluoxetine in offspring rats behavior, 5- serotonin can affect neuronal loop function, so as to explore the pathophysiological mechanism of lactation SSRI antidepressants on offspring the behavior and its possible effects, in order to provide theoretical basis for the study of autism. Methods: the pathogenesis of pregnant SD rats were randomly divided into 2 groups, 3 rats in each group. To be 5-21d after delivery to the rats of the experimental group of fluoxetine (12mg/kg), saline control group rats were given the same amount of normal period lactation. Infants were measured body weight, offspring rats hair, eyes, hearing and free hanging experiment. The offspring born twenty-first days after weaning and male rats were divided into the cage. The childhood rat offspring for burial The food pellet test and open field test (P35d). Childhood and adolescence (P75d) were performed at Western-Blot, IHC, IF content detection of offspring rat prefrontal cortex and dorsal raphe nucleus SERT and TPH. At the same time, the application of ELISA technology in detection of childhood and young adulthood prefrontal cortex and nuclear 5-HT content. The raphe a behavioral test results: 1. body weight: compared with the control group, fluoxetine group weight 9D, 11d, 13D were significantly lower than the control group, there was significant difference (P0.05); 2. hair: compared with the control group, fluoxetine group the length of hair 9D, 11d decreased significantly, no significant difference (P0.05) 13D. In the experimental group decreased significantly compared with the control group, there was significant difference (P0.05): 13D:3. and fluoxetine group compared with the control group had no obvious change in open degree (P0.05); 4. hearing: compared with the control group, fluoxetine group acouesthesia 9D, 11d increased significantly The difference (P0.01).13d experimental group compared with the control group without auditory sensitivity increased significantly, no significant difference (P0.05); 5. buried food pellet test: compared with the control group, fluoxetine group buried food pellet test time 26D had no significant change, no significant difference (P0.05); 6. free hanging experiment: Compared with the control group, fluoxetine group free suspension test at the time of 1ld, 13D were significantly decreased, there was significant difference (P0.01); 7. open field test: compared with the control group, fluoxetine group stool grain number in 28d, 29d, 30d, 31d were significantly increased, there was significant difference (P0.01); fluoxetine stool the average grain number had significant difference (P0.01. Two), Western blot, immunohistochemistry, detect the prefrontal cortex and dorsal raphe nucleus SERT and TPH protein by immunofluorescence method; the content of enzyme-linked immunosorbent assay in the prefrontal cortex and dorsal raphe nucleus 5-HT: 1. Western-blot, IH C, detection of children during the period of IF Technology (35d) and youth (75D) the results showed that SERT protein expression in cortex, raphe nuclei and forehead: compared with lactation saline exposed group, FLX exposure during childhood and adolescence group expression of SERT protein decreased obviously, there was significant difference (P0.05).2. by Western-blot, IHC children, detection technology of IF (35d) and youth (75D) results showed that expression of TPH protein in the cortex, and raphe nuclei: forehead and lactation saline exposed offspring compared to FLX group of children exposed expression of TPH protein decreased obviously, there was significant difference (P0.05).3. IF was used to detect rat prefrontal cortex and the number of axons, results showed that: fluoxetine exposure group children filial mice (35d) and youth (75D) and the number of axons in the prefrontal cortex neurons significantly reduced.4. enzyme-linked immunosorbent assay for detection of 5-HT was found in the prefrontal cortex and the dorsal raphe nucleus and the content: According to groups of childhood (35d) fluoxetine group 5-HT significantly increased total content in the brain, there was significant difference (P0.01); infants (75D) fluoxetine group 5-HT were significantly higher in the prefrontal cortex and dorsal raphe nucleus content, there was significant difference (P0.05). Conclusion]1. lactational exposure to fluoxetine in offsprings the development and abnormal behavior of.2. lactational exposure to fluoxetine by tone generation rat prefrontal cortex and dorsal raphe nucleus in TPH, the expression of SERT protein increased the content of 5-HT.3. exposed to fluoxetine 5-HT content generation in brain tissue increased, may lead to autism behavior class.4. lactational exposure to SSRIs antidepressants in offspring rats behavior learn to have a significant effect, the effect may be caused by 5- serotonergic neuron system disorders.

【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R749.94

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