抑郁模型大鼠海马MKP-1表达及DNA甲基化状态的研究

发布时间：2018-06-25 13:24

本文选题：慢性不可预见性应激 + 抑郁症　；参考：《新乡医学院》2012年硕士论文

【摘要】：目的 1、建立慢性不可预见性应激(CUS)抑郁模型。 2、检测抑郁模型大鼠海马促分裂原活化蛋白激酶磷酸酶-1(MKP-1)表达及启动子区DNA甲基化状态的变化情况。 3、探讨抗抑郁药对抑郁模型大鼠海马MKP-1表达及启动子区DNA甲基化状态的影响。方法 1、动物分组及模型制作选择旷场实验得分相近的30只SD大鼠随机分为3组,正常对照组(A组)10只,生理盐水组(B组)10只,氟西汀组(C组)10只。慢性不可预见性应激共实施42天,每天给予不同的不可预见性应激,一种应激在一周内最多使用2次。用旷场实验和体质量来评价抑郁模型。 2、采用逆转录聚合酶链反应(RT-PCR)方法检测抑郁模型大鼠海马MKP-1mRNA的表达情况。 3、采用联合NaHSO3限制性酶切分析法(COBRA)检测MKP-1基因启动子区DNA甲基化情况。 4、采用统计软件SPSS12.0处理,数据以均数±标准差(x±s)表示,组间比较采用单因素方差分析(ANOVA)。以P0.05为差异有统计学意义。结果 1、旷场试验 (1)水平运动：刺激之前,各组大鼠在水平运动方面差异无统计学意义(P=0.457)。慢性刺激21天后,生理盐水组的水平运动距离远于正常对照组(P=0.005),而生理盐水组与氟西汀组比较差异无统计学意义(P=0.128),正常对照组与氟西汀组比较差异无统计学意义(P=0.148)。药物干预21天后,生理盐水组的水平运动距离比正常对照组水平运动距离缩短(P=0.000),但氟西汀组水平运动距离较生理盐水组延长(P=0.000),正常对照组和氟西汀组的差异无统计学意义(P=0.393)。(2)垂直运动：在垂直运动方面,刺激之前,刺激21天及干预21天时3个时间点正常对照组、生理盐水组与氟西汀组比较差异均无统计学意义(P=0.260,0.960,0.697)。 2、体质量：刺激之前,各组比较差异无统计学意义(P=0.476)。慢性刺激21天时,生理盐水组体质量较正常对照组的体质量减轻(P=0.010),氟西汀组的体质量也较正常对照组的体质量减轻(P=0.000),生理盐水组体质量与氟西汀组体质量比较差异无统计学意义(P=0.109)。干预21天时,生理盐水组体质量较正常对照组的体质量减轻(P=0.000),氟西汀组的体质量也较正常对照组的体质量减轻(PP=0.000),氟西汀组体质量较生理盐水组体质量增加(P=0.026)。 3、生理盐水组MKP-1mRNA的表达较正常对照组升高(P=0.001),较氟西汀组MKP-1mRNA的表达也升高(P=0.024)；但氟西汀组MKP-1mRNA的表达与正常对照组比较差异无统计学意义(P=0.097)。 4、MKP-1基因启动子区DNA甲基化的检测未检测到MKP-1基因启动子区存在DNA甲基化。结论 1、CUS抑郁模型大鼠海马MKP-1mRNA表达水平较正常对照组升高,MKP-1可能参与了抑郁症的发病机制。 2、氟西汀可以明显改善抑郁模型大鼠的抑郁行为,并能改善海马MKP-1mRNA的高表达,提示氟西汀可能通过调节MKP-1mRNA表达参与抗抑郁剂的治疗机制。 3、CUS抑郁模型大鼠海马MKP-1基因的启动子区未发现存在DNA甲基化,提示MKP-1基因启动子区的DNA甲基化可能不参与抑郁症的发病机制。
[Abstract]:objective
1, establish a chronic unpredictable stress (CUS) depression model.
2, we detected the expression of mitogen activated protein kinase phosphatase -1 (MKP-1) and DNA methylation status in hippocampus of depression model rats.
3, to explore the effect of antidepressants on MKP-1 expression and DNA methylation status in hippocampus of depression model rats.
Method
1, 30 SD rats were randomly divided into 3 groups, including 10 rats in the normal control group, 10 in the normal control group, 10 in the saline group (group B) and 10 in the fluoxetine group (group C). The chronic unpredictable stress was carried out for 42 days, and the different unpredictable stress was given every day. A kind of stress was used up to 2 times in a week. Depressive model was evaluated by open field experiment and body mass.
2, reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of MKP-1mRNA in hippocampus of depression model rats.
3, combined NaHSO3 restriction enzyme digestion assay (COBRA) was used to detect the DNA methylation status of MKP-1 gene promoter region.
4, the data were treated with statistical software SPSS12.0, and the data were expressed with mean standard deviation (x + s), and the single factor variance analysis (ANOVA) was used in the group. The difference was statistically significant with the difference of P0.05.
Result
1, open field test
(1) horizontal exercise: before stimulation, there was no significant difference in horizontal movement between the rats in each group (P=0.457). 21 days after the chronic stimulation, the horizontal movement distance of the saline group was far higher than that of the normal control group (P=0.005), while the difference of the normal saline group and the fluoxetine group had no statistical significance (P=0.128), and the normal control group was less than the fluoxetine group. There was no statistical significance (P=0.148). The horizontal movement distance of the normal saline group was shorter than that of the normal control group (P=0.000) after 21 days of drug intervention, but the horizontal movement distance of the fluoxetine group was longer than that of the normal saline group (P=0.000), and there was no significant difference between the normal control group and the fluoxetine group (P=0.393). (2) the vertical movement was in drooping. There was no significant difference between the normal saline group and the fluoxetine group before the stimulation, the stimulation for 21 days and the 3 time points of the intervention for 21 days, and there was no significant difference between the normal saline group and the fluoxetine group (P=0.260,0.960,0.697).
2, body mass: before stimulation, there was no significant difference between each group (P=0.476). At 21 days of chronic stimulation, the mass of the saline group was less than that of the normal control group (P=0.010), and the body mass of the fluoxetine group was also less than that of the normal control group (P=0.000), and the quality of the saline group was compared with the quality of the fluoxetine group. There was no statistical significance (P=0.109). The body mass of the saline group was less than that of the normal control group (P=0.000) at 21 days, and the body mass of the fluoxetine group was also less than that of the normal control group (PP=0.000), and the mass of the fluoxetine group was higher than that of the saline group (P=0.026).
3, the expression of MKP-1mRNA in the normal saline group was higher than that in the normal control group (P=0.001), and the expression of MKP-1mRNA in the fluoxetine group increased (P=0.024), but the expression of MKP-1mRNA in the fluoxetine group was not significantly different from that of the normal control group (P=0.097).
4, detection of DNA methylation in the promoter region of MKP-1 gene did not detect DNA methylation in the promoter region of MKP-1 gene.
conclusion
1, the expression level of MKP-1mRNA in hippocampus of CUS depression model rats is higher than that of normal control group. MKP-1 may be involved in the pathogenesis of depression.
2, fluoxetine can obviously improve the depressive behavior of the depression model rats and improve the high expression of MKP-1mRNA in the hippocampus, suggesting that fluoxetine may be involved in the treatment mechanism of antidepressant by regulating the expression of MKP-1mRNA.
3, DNA methylation was not found in the promoter region of MKP-1 gene in the CUS depression model rats, suggesting that DNA methylation in the promoter region of the MKP-1 gene may not be involved in the pathogenesis of depression.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.4

【参考文献】