舒郁胶囊对抑郁症肝气郁结证模型大鼠海马作用靶点基因筛选及5-HT受体相关基因差异表达的验证

发布时间：2018-04-24 14:00

本文选题：抑郁症肝气郁结证 + 5-羟色胺受体　；参考：《山东中医药大学》2012年硕士论文

【摘要】：目的：用基因芯片检测抑郁症肝气郁结证模型大鼠海马内5-HT受体相关基因的表达变化，初步探讨抑郁症肝气郁结证对海马内5-HT受体的作用机制，并研究调肝方药舒郁胶囊和柴胡皂苷对该病证的干预作用。方法：旷场试验筛选大鼠，采用慢性温和应激法（CMS）复制抑郁症肝气郁结证大鼠模型，模型评价采用糖水偏好实验、旷场试验等。以西药氟西汀为阳性药，并用调肝方药舒郁胶囊和柴胡皂苷进行干预。采用基因芯片检测大鼠海马内差异基因表达情况，比较模型组大鼠、正常组大鼠、舒郁胶囊干预组大鼠、柴胡皂苷干预组大鼠与氟西汀干预组大鼠之间的差异表达基因；用实时荧光定量PCR技术验证5-HT受体相关基因差异表达结果的可靠性。结果：造模刺激4周后，与正常组相比，模型组旷场实验得分有显著性减少（P0.01）。与模型组相比，舒郁组、氟西汀组和柴胡组的得分均有显著性增加(P0.01，P0.001，P0.05)，但三组之间比较没有显著性差异（P0.05）。模型组大鼠糖水摄入量较正常组、舒郁组及氟西汀组显著降低（P0.05，P0.05，P0.01），，说明抑郁症肝气郁结证模型成功，而舒郁胶囊和氟西汀能明显对抗抑郁大鼠糖水摄入量的减少，但两者之间没有显著性差异（P0.05）。柴胡皂苷不能明显改善抑郁大鼠糖水摄入量的减少，但也有一定的作用趋势。采用含有41000个己知基因的大鼠全基因组芯片检测大鼠海马内差异基因表达情况，结果显示模型组、正常组、舒郁组、柴胡组与氟西汀组之间的Htr1a, Htr5b, Htr2c,Htr3a基因存在差异表达。实时荧光定量PCR方法分别检测了Htr1a, Htr5b,Htr2c, Htr3a的表达情况，检测结果与基因芯片的结果基本一致，验证了4个目的基因结果的可靠性。结论：采用CMS法可以成功制备抑郁症肝气郁结证大鼠模型，糖水偏好实验、旷场试验均可评价模型的有效性。各组大鼠海马中Htr1a, Htr5b, Htr2c, Htr3a差异表达，可能是抑郁症肝气郁结证发病的中枢机制之一；中药舒郁胶囊和柴胡皂苷可以有效纠正上述基因表达异常的状态，可能是药物治疗该病症的机制之一。
[Abstract]:Objective: to detect the expression of 5-HT receptor related genes in hippocampus of depression rat model of stagnation of liver-qi by using gene chip, and to explore the mechanism of 5-HT receptor in hippocampus of depression induced by stagnation of liver-qi syndrome. And to study the intervention effect of Shuyu capsule and saikosaponin on the syndrome. Methods: open field test was used to screen rats and chronic mild stress method (CMSs) was used to establish the rat model of depression with stagnation of liver qi. The model was evaluated by sugar water preference test and open field test. Western medicine fluoxetine was used as positive drug, and Shuyu capsule and saikosaponin were used to intervene. The differentially expressed genes in hippocampus of rats were detected by gene chip, and the differentially expressed genes were compared between model group, normal group, Shuyu capsule intervention group, saikosaponin intervention group and fluoxetine intervention group. The reliability of differential expression of 5-HT receptor related genes was verified by real-time fluorescence quantitative PCR. Results: after stimulation for 4 weeks, the scores of open field test in model group were significantly lower than those in normal group (P 0.01). Compared with the model group, the scores of Shuyu group, fluoxetine group and Bupleurum group were significantly increased, but there was no significant difference among the three groups. The intake of sugar water in the model group was higher than that in the normal group, and the intake of sugar water in the Shuyu group and fluoxetine group was significantly lower than that in the control group, indicating that the depression model of stagnation of liver-qi was successful, while Shuyu capsule and fluoxetine could significantly reduce the intake of sugar water in the depressed rats. But there was no significant difference between the two groups (P 0.05). Saikosaponin can not obviously improve the decrease of sugar water intake in depressed rats, but it also has a certain effect trend. The differentially expressed genes of Htr1a, Htr5b and Htr2ct3a were detected by whole genome microarray with 41000 known genes in the hippocampus of rats. The results showed that Htr1a, Htr5b and Htr2ct3a were differentially expressed in model group, normal group, Shuyu group, Chaihu group and fluoxetine group. The expressions of Htr1a, Htr5bHtr2cand Htr3a were detected by real-time fluorescence quantitative PCR. The results were in good agreement with the results of gene chip, and the reliability of the four target gene results were verified. Conclusion: the rat model of depression with stagnation of liver qi, sugar water preference test and open field test can be used to evaluate the validity of the model. The differential expression of Htr1a, Htr5b, Htr2cand Htr3a in hippocampus of rats in each group may be one of the central mechanisms in the pathogenesis of depression due to stagnation of liver-qi, and Shuyu capsule and saikosaponin can effectively correct the abnormal expression of these genes. May be one of the mechanisms of drug treatment of the disease.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285;R-332

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