抑郁心境肝脏代谢酶活性变化及中药干预机制研究

The incidence of depression was rising and associated with a variety of diseases which were stroke, diabetes and other comorbidity. Traditional Chinese medicine (TCM) believed that liver could effect emotion and remove depression with soothing liver. Modern medicine believed that liver could detoxicate, which could transfer the endogenous and exogenous harmful substances to non-toxic or easily excreted substances. Did activity of liver enzyme in depressed mood change significantly? The question had not been studied. Our previous work firstly verified that CYP1A2, CYP2E1 could be strongly induced by depression which came from model of chronic unpredictable stress in rats. CYP1A2 and CYP2E1 are enzymes of “toxicity enhancement”. Modern research showed that depression was related to nerve damage, and some antidepressant drugs would be effective after 2-3 weeks. This study created three different types of models of depression. And the models of depression were evaluated by applying the classical index of depression and target metabolomics method, the activity of CYP450 was evaluated by applying cocktail method, the expression of metabolic enzyme was estimated by applying the molecular biology method. Moreover, the influences of three kinds of famous anti-depressed traditional Chinese medicine on metabolic enzyme in liver were investigated. On this basis, in order to provide new ideas and methods for clinical medical treatment and drug development in depression, the Chinese medicine prescriptions were optimized by molecular docking technology which aiming at key metabolic enzymes, and needed to validate by pharmacological studies.

抑郁症发病率呈上升趋势，且常与中风、糖尿病等多种疾病共病。中医认为肝主情志，疏肝可解郁；现代医学认为肝解毒，可将内源性和外源性有害物质转化为无毒或易于排泄的物质。抑郁心境肝脏代谢酶活性是否发生明显变化？尚未见系统研究。我们前期工作首次证实，大鼠慢性不可预见性应激抑郁模型CYP1A2被强诱导、CYP2E1被中强诱导。CYP1A2和CYP2E1均为“增毒”酶，现代研究表明抑郁症与神经损伤亦有关，有些抗抑郁化学药服用后2-3周才起效。本研究拟建立3种不同类型的抑郁症模型，综合抑郁症经典指标和靶标代谢组学方法评价抑郁症模型，运用Cocktail法评价代谢酶的活性，运用分子生物学方法评价代谢酶的表达。同时考察3种中医治疗抑郁症的著名方剂对肝脏代谢酶活性的影响。在此基础上，针对关键代谢酶，运用分子对接技术优化中药方剂，再经实体药效学实验验证，为抑郁症的临床中医药治疗和新药研发提供新思路和新方法。

本课题采用慢性不可预见性温和应激（CUMS）抑郁模型，探讨抑郁状态是否会对肝脏药物代谢酶产生诱导或抑制作用，在此基础上，给予抗抑郁经典中药方剂半夏厚朴汤，进一步观察其对抑郁状态下的酶活性失调的调节作用。采用鸡尾酒(Cocktail)探针药物法研究CUMS造成的抑郁状态对大鼠体内6种CYP 450亚酶活性的影响。根据Katz法建立CUMS抑郁大鼠模型；分别选用茶碱（CYP1A2）、氯唑沙宗（CYP2E1）、甲苯磺丁脲（CYP2C9）、奥美拉唑（CYP2C19）、右美沙芬（CYP2D6）、咪达唑仑（CYP3A4）作为相应亚酶的探针底物, 采用LC-MS/MS法测定对照组与模型组大鼠体内6种混合探针的血药浓度, 计算药动学参数。结果表明, 茶碱和氯唑沙宗代谢显著加快(P <0.01), 甲苯磺丁脲、右美沙芬、奥美拉唑、咪达唑仑的代谢无显著差异。以上结果说明, 慢性不可预见性温应激造成的抑郁状态对CYP1A2有强诱导、对CYP2E1有中强诱导。采用Cocktail探针药物法评价半夏厚朴汤对CUMS造成的抑郁状态下大鼠体内CYP1A2、2E1亚酶被诱导的调节作用。结果说明，半夏厚朴汤对CUMS模型组大鼠体内CYP1A2和CYP2E1被诱导具有抑制作用。.利血平诱导的慢性抑郁模型状态对大鼠CYP2C6和CYP2D1有中强抑制作用；给予四逆散后对大鼠CYP1A2和2D2有中强抑制作用；给予氟西汀后对大鼠CYP2E1有弱抑制作用。利血平诱导的急性抑郁模型状态对大鼠CYP2D1和CYP2D2有中强抑制作用，对CYP3A2有中强诱导作用；给予四逆散和文拉法辛后对模型大鼠CYP1A2、CYP2C6、CYP2E1、CYP3A2均为中强抑制作用。.采用探针药物法研究肾阳虚抑郁大鼠体内CYP450亚型酶活性变化，及四逆汤的的调节效果。结果表明，肾阳虚抑郁模型大鼠CYP2E1、CYP1A2、CYP2C9被诱导。与模型组比较，各治疗组大鼠血中CYP1A2、CYP2E1、CYP2C6的酶活性逐渐向正常回归，表明四逆汤及氟西汀对肾阳虚抑郁大鼠体内CYP1A2、CYP2E1、CYP2C6的酶活性具有一定的调节作用。