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目的:观察抑郁症模型大鼠海马乙酰胆碱代谢的变化规律及柴胡疏肝散对其影响。方法:60只4-5月龄的SD大鼠随机分为正常对照组(对照组)、模型组、西药组和中药组,共4组,每组15只。利用孤养结合21d慢性不可预见性应激刺激(CUMs)制造抑郁症模型。21d后,采用敞箱实验、糖水消耗量检测和体质量检测等指标评定大鼠行为学改变,并用免疫组织化学染色法观察大鼠大脑海马区乙酰胆碱酯酶(acetylcholinesterase,AChE)与胆碱乙酰化酶(cholineacetyltransferase,ChAT)的蛋白表达变化,应用RT-PCR检测ChAT mRNA的表达,利用比色法检测AChE活性。结果:与对照组相比,模型组水平运动得分、垂直运动得分、体质量和糖水偏爱度均明显降低(P<0.01-0.001),与模型组比较,西药组和中药组水平运动得分、垂直运动得分、糖水偏爱度与体质量均明显升高(P<0.05-0.001);免疫组化结果显示,与对照组相比,模型组海马区ChAT、AChE的蛋白表达均明显降低(P<0.05);RT-PCR结果显示,与对照组相比,模型组大鼠海马区ChAT mRNA表达明显上调(P<0.05),与模型组相比,西药组、中药组海马区ChAT mRNA明显下调(P<0.05);比色法结果显示,与对照组相比,模型组海马区AChE活性明显升高(P<0.05),与模型组相比,西药组、中药组海马区AChE活性明显下降(P<0.05)。结论:胆碱能神经系统与抑郁症发生发展有密切关系。柴胡疏肝散抗抑郁症的作用可能与其抑制大脑组织海马区ChAT蛋白和mRNA表达、降低大脑海马区AChE活性和蛋白表达有关。
Objective: To observe the changes of acetylcholine metabolism in the hippocampus of rats with depression model and the effect of Chaihu Shugan Powder on it. Methods: Sixty Sprague-Dawley (SD) rats aged 4-5 months were randomly divided into normal control group (model group), western medicine group and traditional Chinese medicine group (n = 4). Depression model was established by using solitary combined with 21 d chronic unpredictable stress stimuli (CUMs). After 21 days, behavioral changes were assessed by open-field test, sugar consumption test and body weight test, and immunohistochemical staining was used to observe the changes of acetylcholinesterase (AChE) and choline acetylation The protein expression of Choline acetyltransferase (ChAT) was detected. The expression of ChAT mRNA was detected by RT-PCR and the activity of AChE was detected by colorimetric assay. Results: Compared with the control group, the scores of horizontal movement, vertical movement, body weight and sugar preference of the model group were significantly decreased (P <0.01-0.001). Compared with the model group, the scores of horizontal movement, vertical (P <0.05-0.001). The results of immunohistochemistry showed that compared with the control group, the protein expression of ChAT and AChE in hippocampus of model group were significantly decreased (P <0.05 (P <0.05). Compared with the model group, the expression of ChAT mRNA in the hippocampus of the model group was significantly lower than that in the control group (P <0.05) <0.05). Compared with the control group, AChE activity in hippocampus of model group was significantly increased (P <0.05) by colorimetry. Compared with model group, AChE activity in hippocampus of western medicine group and traditional Chinese medicine group decreased significantly (P < <0.05). Conclusion: The cholinergic nervous system is closely related to the occurrence and development of depression. The effect of Chaihu Shugan San on depression may be related to the inhibition of the expression of ChAT protein and mRNA in the hippocampus and the decrease of AChE activity and protein expression in the hippocampus.