培元解郁方对慢性应激所致抑郁症的抗抑郁作用及TRP-KYN代谢途径的调节

发布时间：2018-04-30 00:20

本文选题：应激 + TRP—KYN代谢　；参考：《北京中医药大学》2017年硕士论文

【摘要】：随着社会节奏的改变,抑郁症发病率逐年攀升,据世界卫生组织(WTO)预测,到2020年抑郁症将成为世界第二大医疗疾患。抑郁症严重影响着患者的身心健康并给家庭和社会带来沉重的负担。因此,关于抑郁症发病机制和治疗药物的研究十分迫切。目前有关抑郁症的发病机制存在几种假说,包括细胞因子假说,下丘脑-垂体-肾上腺皮质(HPA)轴假说,单胺能假说以及神经可塑性假说等。目前,常用的抗抑郁药物有单胺氧化酶抑制剂、三环类抑郁剂、5-羟色胺再摄取抑制剂和5-羟色胺去甲肾上腺素再摄取抑制剂。但是,西药治疗抑郁症存在很多问题,包括治愈率低、副作用大以及依从性差等,所以人们将焦点逐渐转向传统中医药领域。近年来色氨酸-犬尿氨酸(TRP-KYN)代谢异常逐渐成为抑郁症发病机制研究的切入点。色氨酸是神经递质五羟色胺(5-HT)的合成原料,在色氨酸羟化酶作用下合成5-HT,而在IDO或TDO的作用下,通过TRP-KYN代谢途径,形成犬尿氨酸等活性产物。当TRP-KYN代谢发生异常时,色氨酸耗竭,造成5-HT合成降低,继而引发抑郁症。本实验以这一代谢为切入点探讨培元解郁方的抗抑郁作用以及对TRP-KYN代谢通路的调节作用。1.培元解郁方对慢性应激BALB/c小鼠的抗抑郁作用及TRP—KYN代谢途径的调节目的:研究四逆散、天丝饮和培元解郁方的抗抑郁作用以及对血清皮质酮含量和TRP-KYN代谢途径的关键酶色氨酸2,3-双加氧酶(TDO)的影响。方法:束缚加噪声5d制备BALB/c小鼠抑郁模型,用Noldus EthoVision XT9系统采集分析小鼠强迫游泳不动时间,采用HPLC-MS/MS技术检测血清皮质酮(C)、色氨酸(TRP)、犬尿氨酸(KYN)、喹啉酸(Q)、犬尿烯酸(KA)的含量,实时荧光定量PCR检测肝组织中TDOmRNA表达水平。结果:束缚噪声应激可以引起模型组小鼠强迫游泳不动时间延长,血清皮质酮含量增加,KYN/TRP、KYN/KA比值升高,KYN/Q比值降低。四逆散、天丝饮、培元解郁方能缩短小鼠游泳不动时间(P0.01),降低血清皮质酮含量(P0.05),提高KYN/TRP、KYN/Q比值(P0.05),降低KYN/KA比值。结论:四逆散、天丝饮、培元解郁方都可以降低慢性应激BALB/c小鼠强迫游泳不动时间,对抗应激引起的血清皮质酮含量升高,抑制TRP-KYN代谢途径中的神经毒性代谢产物Q产生。其中培元解郁方、天丝饮缩短强迫游泳不动时间效果更显著;四逆散下调血清皮质酮含量作用更明显,培元解郁方可以促进神经保护作用产物KA的生成。因此,三组方均有抗抑郁作用,而培元解郁方在缩短游泳不动时间、调节TRP-KYN代谢方面效果更明显。2.培元解郁方对糖皮质激素给药大鼠的抗抑郁作用及对TRP—KYN代谢途径的调节目的:研究天丝饮、四逆散以及不同剂量培元解郁方对糖皮质激素给药大鼠的抗抑郁作用,对TRP-KYN代谢途径关键酶TDO和IDO表达的影响以及对海马和皮层神经元可塑性的影响。方法:糖皮质激素注射28d建立大鼠抑郁模型,造模结束后采用敞箱测试和糖水消耗测试观察天丝饮、四逆散和培元解郁方对抑郁大鼠行为的改善情况;采用HPLC-MS/MS技术检测血清TRP、KYN、KA、Q含量,RT-PCR检测肝组织TDO,海马和皮层 IDO、CREB、NMDA、BDNF、TrkB、GR、5-HT1A mRNA 表达。结果:1.糖水测试结果:与对照组相比,模型组大鼠糖水消耗量显著降低(p0.05),与模型组比较和厚朴酚、培元解郁高剂量组大鼠糖水消耗量显著升高(p0.05)。2.敞箱测试结果:与空白组比较,模型组大鼠敞箱活动的总得分显著降低(p0.05),与模型组比较,氟西汀、天丝饮、培元解郁方中剂量和培元解郁方高剂量组大鼠敞箱活动总得分显著升高(p0.05)。以上行为学测试表明培元解郁方可以改善糖皮质激素给药大鼠的抑郁行为。3.TRP—KYN代谢:与空白组比较,模型组KYN/TRP、KYN/KA比值显著升高(p0.05),IDO和TDO mRNA表达降低(p0.05),与模型组比较培元解郁高剂量组KYN/TRP、KYN/KA比值显著降低(p0.05),KYN/Q比值显著升高(p0.05),培元解郁方中剂量组KYN/TRP比值显著降低(p0.05),氟西汀、天丝饮组KYN/KA比值显著降低(p0.05)。除和厚朴酚外,各给药组TDO mRNA表达显著升高(p0.05)。4.皮层、海马神经元可塑性:与空白组比较,模型组海马BDNF、5-HT1A受体、皮层TrkB受体mRNA表达显著降低(p0.05),皮层NMDA、GR受体mRNA表达显著升高(p0.05)。与模型组比较,培元解郁方中、高剂量组海马BDNF、5-HT1A表达显著升高(p0.05),皮层GR、NMDA受体表达显著降低(p0.05),培元解郁方高剂量组皮层TrkB表达显著升高(p0.05)。结论:培元解郁方高剂量能提高敞箱活动、增加糖水消耗量,天丝饮仅增加糖水消耗,四逆散对敞箱活动和糖水消耗作用不明显;培元解郁方可以下调TRP—KYN代谢,抑制神经毒性产物Q产生、促进神经保护作用产物KA产生,天丝饮仅促进KA产生,四逆散对TRP-KYN代谢的调节作用不明显;培元解郁方可以降低皮层GR、NMDA受体表达,提高海马和皮层BDNF、5-HT1A表达;四逆散和天丝饮对海马作用不明显,对于皮层的作用与培元解郁方类似,但四逆散提高皮层BDNF表达作用不明显。综上,培元解郁方可以通过下调TRP—KYN代谢,抑制神经毒性,提高神经保护,促进皮层和海马,尤其是皮层的神经元可塑性发挥抗抑郁作用。
[Abstract]:With the change of social rhythm, the incidence of depression is increasing year by year. According to the WHO (WTO), depression will become the second largest medical disease in the world by 2020. Depression seriously affects the physical and mental health of the patients and brings heavy burden to the family and society. Therefore, research on the pathogenesis of depression and the treatment of drugs is ten. There are several hypotheses about the pathogenesis of depression, including the cytokine hypothesis, the hypothalamus pituitary adrenal cortex (HPA) axis hypothesis, the monoamine hypothesis and the neural plasticity hypothesis. At present, the commonly used antidepressants are monoamine oxidase inhibitor, tricyclic depressant, 5- serotonin reuptake inhibitor and 5- hydroxyl. However, there are many problems in the treatment of depression in western medicine, including low cure rate, large side effects and poor compliance, so people gradually turn to the field of traditional Chinese medicine. In recent years, the abnormal metabolism of tryptophan - canine urinary ammonia (TRP-KYN) has gradually become a breakthrough in the study of the pathogenesis of depression. Point. Tryptophan is a synthetic raw material of neurotransmitter five hydroxytryptamine (5-HT), synthesized 5-HT under the action of tryptophan hydroxylase, and formed the active product of canine urinary ammonia by TRP-KYN metabolism pathway under the action of IDO or TDO. When the metabolism of TRP-KYN is abnormal, the depletion of tryptophan causes the decrease of 5-HT synthesis and then causes depression. This experiment is based on this experiment. The antidepressant effect of Pei Yuan Jie Yu recipe and the regulation of TRP-KYN metabolic pathway on the antidepressant effect of.1. and the regulation of TRP KYN metabolic pathway on chronic stress BALB/c mice were studied. The antidepressant effect and the content of serum corticosterone in the four reverse powder, tiun Yin and Pei Yuan Jie Yu Fang were studied. The effects of the key enzyme tryptophan 2,3- double oxygenase (TDO) on the metabolic pathway of TRP-KYN. Methods: binding and noise 5D were used to prepare the BALB/c mice depression model. The time of forced swimming in mice was collected and analyzed with Noldus EthoVision XT9 system. The serum corticosterone (C), tryptophan (TRP), canine urinary ammonia acid (KYN), quinoline acid, dog were detected by Noldus EthoVision XT9 system. The content of urinary enuric acid (KA) and real-time fluorescence quantitative PCR were used to detect the expression level of TDOmRNA in liver tissue. Results: restraint noise stress can cause prolonged swimming time, increase of serum corticosterone content, increase of KYN/TRP, KYN/KA ratio, and lower KYN/Q ratio in model mice. Four inverse scatter, Tien Yin and Pei Yuan Jie Yu can shorten the swimming of mice. Time (P0.01), lower serum corticosterone (P0.05), increase the ratio of KYN/TRP, KYN/Q (P0.05), and reduce the ratio of KYN/KA. Conclusion: four inverse scatter, Tien Yin and Pei Yuan Jie Yu Fang can reduce the time of forced swimming in BALB/c mice with chronic stress, the increase of serum corticosterone in the anti stress induced serum and the inhibition of neurotoxicity in the metabolic pathway of TRP-KYN Q was produced by the product of sexual metabolism. Among them, the effect of shortening the time of forced swimming was more significant, and the effect of four inverse scatter on the content of serum corticosterone was more obvious, and the prescription of Pei Yuan Jie Yu could promote the production of KA in the product of neuroprotective effect. The antidepressant effect of.2. Pei Yuan Jie Yu Fang on the rats with glucocorticoid administration and the regulation of TRP KYN metabolic pathway were more obvious in regulating the metabolism of TRP-KYN: the antidepressant effect of Tiantian decoction, four inverse scatter and the different doses of Pei Yuan Jie Yu prescription on the rats with glucocorticoid, and the key enzyme of TRP-KYN metabolic pathway TDO and IDO table The effect of DA on the plasticity of hippocampal and cortical neurons. Methods: the rat model of depression was established by injection of glucocorticoid with 28d. After the model, the open box test and sugar water consumption test were used to observe the Tiantian drink. The behavior improvement of the depressive rats was improved by the four inverse scatter and the Pei Yuan Jie Yu Fang, and the serum TRP, KYN, KA were detected by HPLC-MS/MS technique. Q content, RT-PCR detection of liver tissue TDO, hippocampus and cortex IDO, CREB, NMDA, BDNF, TrkB, GR, 5-HT1A mRNA expression. Results: 1. sugar water test results: compared with the control group, the sugar water consumption in the model group was significantly lower (P0.05), compared with the model group and the magnolol and the high dose group, the sugar water consumption was significantly increased. Test results: compared with the blank group, the total score of the open box activity in the model group was significantly lower (P0.05). Compared with the model group, the total score of open box activity in the high dose group was significantly increased (P0.05) in the high dose group of fluoxetine, Tiantian drink, Pei Yuan Jie Yu Fang and Pei Yuan Jie Yu Fang. The above behavioral test showed that the corticosteroid could improve the glucocorticoid .3.TRP - KYN metabolism in the depressive behavior of the rats: compared with the blank group, the ratio of KYN/TRP and KYN/KA in the model group was significantly increased (P0.05), the expression of IDO and TDO mRNA decreased (P0.05), and the ratio of KYN/TRP in the high dose group was significantly lower than that of the model group, and the KYN/KA ratio was significantly reduced (P0.05), the ratio was significantly higher than that in the model group. The value of KYN/KA in fluoxetine and Tian Yin group decreased significantly (P0.05). Except for the magnolol and Magnolia, the expression of TDO mRNA was significantly increased (P0.05).4. cortex and the plasticity of hippocampal neurons: compared with the blank group, the mRNA expression of BDNF, 5-HT1A receptor and TrkB receptor in the model group was significantly reduced (P0.05). The expression of BDNF and 5-HT1A in the hippocampus of high dose group increased significantly (P0.05), and the expression of GR and NMDA receptors in the cortex was significantly decreased (P0.05), and the expression of TrkB in the high dose group of Pei Yuan Jie Yu Fang was significantly increased (P0.05). Conclusion: the high dose of Pei Yuan Jie Yu Fang can increase the open box activity and increase the consumption of sugar water. It only increased the consumption of sugar and water, and the effect of four inverse scatter on open box activity and sugar water consumption was not obvious; Pei Yuan Jie Yu Fang could reduce the metabolism of TRP - KYN, inhibit the production of neurotoxic product Q, promote the production of neuroprotective product KA, and promote the production of KA only. The regulation effect of four inverse powder on the metabolism of TRP-KYN is not obvious. In order to reduce the expression of GR and NMDA receptor in the cortex and the expression of BDNF and 5-HT1A in the hippocampus and cortex, the effect of four inverse scatter and Tian silk drink on the hippocampus is not obvious, and the effect on the cortex is similar to that of the Pei Yuan Jie Yu Fang, but the four inverse dispersion improves the expression of BDNF in the cortex. Neuroprotection promotes neuronal plasticity in the cortex and hippocampus, especially in the cortex, and plays an antidepressant role.

【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】