柴胡疏肝散对抑郁模型大鼠海马乙酰胆碱代谢的影响

Effect of Chaihu Shugan Powderon metabolism of ach in brain hippocampus tissue of depressive rats

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 of hippocampal of depression models on rats and the effect of Chaihu Shugan Powder on it.Method： 60 male Sprague-Dawley rats of 4-5 month were randomly divided into four groups and 15 rats in 4 groups： normal control group,model group,western medicine group and Chinese medicine group.The depressed modle was reduced by single feed combined with CUMS.After twenty-one days,behavior changes in rats were detected through open-field test,sugar consumption test and body weight;expression changes of acetylcholine transferase and acetylcholinesterase in hippocampus of rats brain were observed with immunochemistry method;ChAT mRNA expression was detected with RT-PCR;the activity of AChE in hippocampus of rats brain tissue was measured with colorimetric method.Results： On the 21st day of model replication,compared with normal control group,the scores of horizontal activity and vertical activity,weight and sugar preference degrees in model group decreased significantly（P0.01-0.001）;Compared with model group,western medicine group and Chinese medicine group got significantly higher scores of horizontal activity and vertical activity,weight and sugar preference degrees（P0.05-0.001）.The results of immunochemistry staining showed that ChAT,AChEprotein expression was significantly increased in hippocampus of model group compared with that of normal control group（P0.05）;Compared with model group,western medicine group and Chinese medicine group had significantly lower ChAT,AChE protein expression in hippocampus（P0.05）.RT-PCR results showed that expression levels of ChAT mRNA in hippocampus of rats of model group were significantly raised compared with those of normal control group（P0.05）;Compared with those of model group,expression levels of ChAT mRNA in hippocampus of rats of western medicine group and Chinese medicine group was significantly reduced（P0.05）;The results of colorimetric method showed that AChE activity in hippocampus of model group was significantly increased compared with that of normal control group（P0.05）;Compared with that of model group,AChE activity in hippocampus of western medicine group and Chinese medicine group was significantly decreased（P0.05）.Conclusion： Cholinergic nervous system is closely related to the occurrence and development of depression.Antidepressant function of Chaihu Shugan Powder may be related to its inhibition to the protein and mRNA expression of ChAT,and to reducing the activity and protein expression of AChE.