青春期雌性小鼠抑郁模型建立及行为学研究与药物干预

发布时间：2018-08-19 20:28

【摘要】：研究目的:青春期抑郁症具有其特殊性,青春期前男孩和女孩的抑郁症发病率无明显差别;但进入青春期后,女孩抑郁症发病率急剧增加,约为男孩发病的2-3倍。但目前抑郁症的研究多集中于成年人群和雄性啮齿类动物,针对青春期女性抑郁症的相关研究尚不丰富,探讨青春期女性抑郁症的有效防治途径具有深远的学术意义与临床价值。本论文在制备青春期雌性小鼠抑郁模型基础上,应用具有快速抗抑郁效果的氯胺酮和GLYX-13两种药物,通过行为学研究观察对青春期雌性抑郁小鼠的抑郁样行为和空间记忆能力的改善作用。研究方法:1.建立青春期雌性小鼠抑郁模型C57BL/6J 18天雌性、雄性小鼠各16只,分别随机分为:雌性对照组、雌性模型组、雄性对照组和雌性模型组,每组各8只。采用慢性轻度不可预知应激模型(chronic unpredictable mild stress,CUMS)方法建立抑郁模型,采用水平震荡、束缚、噪声、禁食、禁水、昼夜颠倒、潮湿垫料、空笼饲养、倾斜鼠笼、陌生鼠笼等10种应激源,模型组每天随机不重复给予2种应激持续3周,且程度不同的应激源每周相应增加。3周期间每天观察青春期雌性小鼠的阴道口开放状态,3周后观察比较4组小鼠的体质量变化、糖水偏爱测试的糖水消耗百分数、旷场实验中的自主活动能力、运动距离和中央区时间、高架十字迷宫实验的开放臂时间、悬尾实验和强迫游泳实验的不动时间来判断是否出现抑郁样行为。2.氯胺酮和GLYX-13对青春期雌性小鼠抑郁样行为的药物干预C57BL/6J 18天雌性小鼠39只,随机分为生理盐水对照组(Con+NS)、生理盐水模型组(CUMS+NS)、氯胺酮对照组(Con+Ket)、氯胺酮干预组(CUMS+Ket)、GLYX-13对照组(Con+GLYX-13)和GLYX-13干预组(CUMS+GLYX-13)6组,对照组每组各6只,模型组和干预组每组各7只。比较制备3周CUMS后,各组小鼠药物干预前后的自由活动能力。通过各组小鼠旷场实验的运动距离和中央区时间、高架十字迷宫实验的开放臂时间、悬尾和强迫游泳实验的不动时间判断是否表现抑郁样行为。通过抑郁样行为的改善程度比较造模后药物干预前后各模型组小鼠的变化情况。3.青春期雌性抑郁小鼠空间记忆能力的研究主动逃避实验可以通过小鼠在转盘上的行为表现探索空间记忆能力、认知协调能力和认知灵活性。实验动物分组同上2中所述,通过各组小鼠主动逃避实验的运动轨迹、第一次进入带电区域的时间和进入带电区域的次数来判断小鼠的空间记忆能力。通过空间记忆能力的提高程度比较造模后药物干预前后各模型组小鼠的改善效果。研究结果:1.3周CUMS后,雌性模型组较雌性对照组的自主活动能力下降,体质量变化、糖水消耗百分数、运动距离、中央区时间、开放臂时间、悬尾和强迫游泳不动时间均有不同程度的统计学意义(P0.05或P0.01或P0.001);雄性模型组较雄性对照组的体质量变化、糖水消耗百分数、运动距离、中央区时间、强迫游泳不动时间均有不同程度的统计学意义(P0.05或P0.01或P0.001);雌性模型组较雄性模型组的自主活动能力下降更明显,其运动距离、开放臂时间、悬尾和强迫游泳不动时间均有不同程度的统计学意义(P0.05或P0.01)。2.3周CUMS后,各模型组较相应对照组的自主活动能力均下降,糖水偏爱测试、旷场实验、高架十字迷宫实验、悬尾实验和强迫游泳实验等各项行为学实验结果有统计学意义(P0.05)。给药后,氯胺酮干预组和GLYX-13干预组的自主活动能力均较给药前改善,氯胺酮干预组与氯胺酮对照组的自主活动能力无明显差异。氯胺酮干预组和GLYX-13干预组与各对照组比较各项结果均无统计学意义(P0.05);氯胺酮干预组较生理盐水模型组的运动距离和悬尾不动时间有统计学意义(P0.05);GLYX-13干预组较生理盐水模型组强迫游泳不动时间有统计学意义(P0.05)。3.3周CUMS后,主动逃避实验中,Trial 1的运动轨迹显示,模型组较相应对照组空间记忆能力下降。Trial 3各模型组较生理盐水对照组第一次进入带电区域的时间缩短有统计学意义(P0.05);Trial 6生理盐水模型组和GLYX-13干预组较不同对照组进入带电区域的次数有不同程度的统计学意义(P0.05或P0.01)。给药后,Trial 1的运动轨迹显示,氯胺酮干预组和GLYX-13干预组的空间记忆能力改善不明显。Trial 1氯胺酮干预组较生理盐水模型组和GLYX-13干预组第一次进入带电区域的时间延长有统计学意义(P0.05或P0.01);Trial1和Trial 6中,氯胺酮干预组和GLYX-13干预组较生理盐水模型组进入带电区域的次数减少有统计学意义(P0.05)。结论:1.青春期雌性、雄性小鼠均造模成功,但雌性模型组的行为学抑郁样表现比雄性组更加明显,相同应激对青春期雌性小鼠的影响较雄性小鼠更加严重。雌性模型组与雄性模型组相比对CUMS更加易感,抑郁程度更明显。2.氯胺酮和GLYX-13具有快速抗抑郁作用,均可改善青春期雌性抑郁小鼠的抑郁样表现,在不同实验中的改善程度不同。3.氯胺酮和GLYX-13均可改善青春期雌性抑郁小鼠的空间记忆能力,但氯胺酮较GLYX-13的改善作用更快。
[Abstract]:Objectives: Adolescent depression has its particularity, and there is no significant difference in the incidence of depression between pre-adolescent boys and girls; however, after puberty, the incidence of depression in girls increases dramatically, about 2-3 times as much as in boys. However, most of the current studies on depression focus on adult populations and male rodents, targeting adolescent women. The related research of depression is not abundant, and it has far-reaching academic significance and clinical value to explore the effective prevention and treatment of adolescent female depression. Based on the preparation of adolescent female mice depression model, this paper applied ketamine and GLYX-13 with rapid antidepressant effect to observe the effect of adolescence through behavioral research. Methods: 1. Establishment of depression model C57BL/6J 18 days female mice and 16 male mice were randomly divided into female control group, female model group, male control group and female model group with 8 mice in each group. Chronic unpredictable mild stress (CUMS) was used to establish depression model. Ten stress sources, including horizontal oscillation, restraint, noise, fasting, water-fast, day-night inversion, wet mat, empty cage feeding, tilted cage, unfamiliar cage, were used. The model group was given two kinds of stress without repetition every day for three weeks, with different degrees of stress sources weekly phase. The open state of the vaginal orifice in adolescent female mice should be observed daily for 3 weeks, and the body mass changes, the percentage of sugar consumption in the sugar preference test, the autonomous activity in the open field test, the distance of movement and the time of the central area, the open arm time in the elevated cross-maze test, the tail suspension test and the strength of the four groups were observed and compared after 3 weeks. Drug intervention of ketamine and GLYX-13 on depression-like behavior in adolescent female mice C57BL/6J 18 days female mice were randomly divided into normal saline control group (Con+NS), normal saline model group (CUMS+NS), ketamine control group (Con+Ket), ketamine intervention group (CUMS+NS). Ket, GLYX-13 control group (Con+GLYX-13) and GLYX-13 intervention group (CUMS+GLYX-13) 6 groups, 6 in each control group, 7 in each model group and 7 in each intervention group. After three weeks of preparation of CUMS, the free activity of mice in each group before and after drug intervention was compared. The time of open arms, tail suspension and immobile time of forced swimming test were used to determine whether depression-like behavior was present. The changes of mice in each model group before and after drug intervention were compared by the improvement of depression-like behavior. Behavioral performance explores spatial memory, cognitive coordination, and cognitive flexibility. The experimental animals were grouped into the same groups as described in the previous 2. The spatial memory of the mice was judged by the locus of movement of each group, the time of first entering the charged area and the number of times of entering the charged area. Results: After 1.3 weeks of CUMS, the autonomic activity, body weight, percentage of sugar consumption, exercise distance, central area time, open arm time, tail suspension and forced swimming immobility time of the female model group were all decreased compared with those of the female control group. Statistical significance (P 0.05 or P 0.01 or P 0.001); body mass change, sugar consumption percentage, exercise distance, central area time, forced swimming immobility time in male model group compared with male control group were statistically significant (P 0.05 or P 0.01 or P 0.001); After 2.3 weeks of CUMS, the autonomic activity of each model group was decreased compared with that of the corresponding control group, including sugar preference test, open field test, elevated cross maze test, tail suspension test and forced swimming test. There was no significant difference between the ketamine intervention group and the ketamine control group. There was no significant difference between the ketamine intervention group and the GLYX-13 intervention group. Meaning (P 0.05); Ketamine intervention group than normal saline model group movement distance and tail suspension immobility time were statistically significant (P 0.05); GLYX-13 intervention group than normal saline model group forced swimming immobility time was statistically significant (P 0.05). 3.3 weeks after CUMS, the active escape experiment Trial 1 locus showed that the model group compared with the corresponding control group. The time of first entering the charged area in Trial-3 model group was shorter than that in normal saline control group (P 0.05), and the times of entering the charged area in Trial-6 model group and GLYX-13 intervention group were different from those in different control group (P 0.05 or P 0.01). The movement track showed that the spatial memory ability of the ketamine intervention group and GLYX-13 intervention group was not significantly improved. The time of the first entry into the charged area of the Trial-1 ketamine intervention group was significantly longer than that of the normal saline model group and GLYX-13 intervention group (P 0.05 or P 0.01); Trial-1 and Trial-6, ketamine intervention group and GLYX-13 intervention group were significantly longer than that of the normal saline model group and GLYX-13 intervention group (P 0.05 or P 0.01). Conclusion: 1. The adolescent female and male mice were successful in modeling, but the behavioral depression in the female model group was more obvious than that in the male model group. The same stress had more serious influence on the adolescent female mice than the male mice. Ketamine and GLYX-13 have a rapid antidepressant effect, both can improve the depression-like performance of adolescent female depressive mice, and the improvement degree is different in different experiments. 3. Ketamine and GLYX-13 can improve the spatial memory ability of adolescent female depressive mice, but ketamine can improve the spatial memory ability of adolescent female depressive mice. Ketones improved faster than GLYX-13.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4;R-332

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