本文選題：應激 + TRP—KYN代謝　； 參考：《北京中醫(yī)藥大學》2017年碩士論文

【摘要】：隨著社會節(jié)奏的改變,抑郁癥發(fā)病率逐年攀升,據(jù)世界衛(wèi)生組織(WTO)預測,到2020年抑郁癥將成為世界第二大醫(yī)療疾患。抑郁癥嚴重影響著患者的身心健康并給家庭和社會帶來沉重的負擔。因此,關于抑郁癥發(fā)病機制和治療藥物的研究十分迫切。目前有關抑郁癥的發(fā)病機制存在幾種假說,包括細胞因子假說,下丘腦-垂體-腎上腺皮質(HPA)軸假說,單胺能假說以及神經(jīng)可塑性假說等。目前,常用的抗抑郁藥物有單胺氧化酶抑制劑、三環(huán)類抑郁劑、5-羥色胺再攝取抑制劑和5-羥色胺去甲腎上腺素再攝取抑制劑。但是,西藥治療抑郁癥存在很多問題,包括治愈率低、副作用大以及依從性差等,所以人們將焦點逐漸轉向傳統(tǒng)中醫(yī)藥領域。近年來色氨酸-犬尿氨酸(TRP-KYN)代謝異常逐漸成為抑郁癥發(fā)病機制研究的切入點。色氨酸是神經(jīng)遞質五羥色胺(5-HT)的合成原料,在色氨酸羥化酶作用下合成5-HT,而在IDO或TDO的作用下,通過TRP-KYN代謝途徑,形成犬尿氨酸等活性產(chǎn)物。當TRP-KYN代謝發(fā)生異常時,色氨酸耗竭,造成5-HT合成降低,繼而引發(fā)抑郁癥。本實驗以這一代謝為切入點探討培元解郁方的抗抑郁作用以及對TRP-KYN代謝通路的調節(jié)作用。1.培元解郁方對慢性應激BALB/c小鼠的抗抑郁作用及TRP—KYN代謝途徑的調節(jié)目的:研究四逆散、天絲飲和培元解郁方的抗抑郁作用以及對血清皮質酮含量和TRP-KYN代謝途徑的關鍵酶色氨酸2,3-雙加氧酶(TDO)的影響。方法:束縛加噪聲5d制備BALB/c小鼠抑郁模型,用Noldus EthoVision XT9系統(tǒng)采集分析小鼠強迫游泳不動時間,采用HPLC-MS/MS技術檢測血清皮質酮(C)、色氨酸(TRP)、犬尿氨酸(KYN)、喹啉酸(Q)、犬尿烯酸(KA)的含量,實時熒光定量PCR檢測肝組織中TDOmRNA表達水平。結果:束縛噪聲應激可以引起模型組小鼠強迫游泳不動時間延長,血清皮質酮含量增加,KYN/TRP、KYN/KA比值升高,KYN/Q比值降低。四逆散、天絲飲、培元解郁方能縮短小鼠游泳不動時間(P0.01),降低血清皮質酮含量(P0.05),提高KYN/TRP、KYN/Q比值(P0.05),降低KYN/KA比值。結論:四逆散、天絲飲、培元解郁方都可以降低慢性應激BALB/c小鼠強迫游泳不動時間,對抗應激引起的血清皮質酮含量升高,抑制TRP-KYN代謝途徑中的神經(jīng)毒性代謝產(chǎn)物Q產(chǎn)生。其中培元解郁方、天絲飲縮短強迫游泳不動時間效果更顯著;四逆散下調血清皮質酮含量作用更明顯,培元解郁方可以促進神經(jīng)保護作用產(chǎn)物KA的生成。因此,三組方均有抗抑郁作用,而培元解郁方在縮短游泳不動時間、調節(jié)TRP-KYN代謝方面效果更明顯。2.培元解郁方對糖皮質激素給藥大鼠的抗抑郁作用及對TRP—KYN代謝途徑的調節(jié)目的:研究天絲飲、四逆散以及不同劑量培元解郁方對糖皮質激素給藥大鼠的抗抑郁作用,對TRP-KYN代謝途徑關鍵酶TDO和IDO表達的影響以及對海馬和皮層神經(jīng)元可塑性的影響。方法:糖皮質激素注射28d建立大鼠抑郁模型,造模結束后采用敞箱測試和糖水消耗測試觀察天絲飲、四逆散和培元解郁方對抑郁大鼠行為的改善情況;采用HPLC-MS/MS技術檢測血清TRP、KYN、KA、Q含量,RT-PCR檢測肝組織TDO,海馬和皮層 IDO、CREB、NMDA、BDNF、TrkB、GR、5-HT1A mRNA 表達。結果:1.糖水測試結果:與對照組相比,模型組大鼠糖水消耗量顯著降低(p0.05),與模型組比較和厚樸酚、培元解郁高劑量組大鼠糖水消耗量顯著升高(p0.05)。2.敞箱測試結果:與空白組比較,模型組大鼠敞箱活動的總得分顯著降低(p0.05),與模型組比較,氟西汀、天絲飲、培元解郁方中劑量和培元解郁方高劑量組大鼠敞箱活動總得分顯著升高(p0.05)。以上行為學測試表明培元解郁方可以改善糖皮質激素給藥大鼠的抑郁行為。3.TRP—KYN代謝:與空白組比較,模型組KYN/TRP、KYN/KA比值顯著升高(p0.05),IDO和TDO mRNA表達降低(p0.05),與模型組比較培元解郁高劑量組KYN/TRP、KYN/KA比值顯著降低(p0.05),KYN/Q比值顯著升高(p0.05),培元解郁方中劑量組KYN/TRP比值顯著降低(p0.05),氟西汀、天絲飲組KYN/KA比值顯著降低(p0.05)。除和厚樸酚外,各給藥組TDO mRNA表達顯著升高(p0.05)。4.皮層、海馬神經(jīng)元可塑性:與空白組比較,模型組海馬BDNF、5-HT1A受體、皮層TrkB受體mRNA表達顯著降低(p0.05),皮層NMDA、GR受體mRNA表達顯著升高(p0.05)。與模型組比較,培元解郁方中、高劑量組海馬BDNF、5-HT1A表達顯著升高(p0.05),皮層GR、NMDA受體表達顯著降低(p0.05),培元解郁方高劑量組皮層TrkB表達顯著升高(p0.05)。結論:培元解郁方高劑量能提高敞箱活動、增加糖水消耗量,天絲飲僅增加糖水消耗,四逆散對敞箱活動和糖水消耗作用不明顯;培元解郁方可以下調TRP—KYN代謝,抑制神經(jīng)毒性產(chǎn)物Q產(chǎn)生、促進神經(jīng)保護作用產(chǎn)物KA產(chǎn)生,天絲飲僅促進KA產(chǎn)生,四逆散對TRP-KYN代謝的調節(jié)作用不明顯;培元解郁方可以降低皮層GR、NMDA受體表達,提高海馬和皮層BDNF、5-HT1A表達;四逆散和天絲飲對海馬作用不明顯,對于皮層的作用與培元解郁方類似,但四逆散提高皮層BDNF表達作用不明顯。綜上,培元解郁方可以通過下調TRP—KYN代謝,抑制神經(jīng)毒性,提高神經(jīng)保護,促進皮層和海馬,尤其是皮層的神經(jīng)元可塑性發(fā)揮抗抑郁作用。
[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.

【學位授予單位】：北京中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5

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本文編號：1822223