本文選題：芍藥苷 + 芍藥內(nèi)酯苷　； 參考：《北京中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：背景血虛肝郁證是肝血虧虛所引起的肝失疏泄調(diào)達(dá),既有肝經(jīng)血分的虧虛,又有肝經(jīng)氣機(jī)的疏泄失常,是肝血虧虛和肝失疏泄并存的復(fù)合中醫(yī)證候。中醫(yī)學(xué)肝藏血功能與現(xiàn)代醫(yī)學(xué)肝臟在物質(zhì)和能量代謝的作用有著密切的關(guān)系,均可參與神經(jīng)-內(nèi)分泌-免疫(NEI)網(wǎng)絡(luò)中肽類激素、免疫細(xì)胞因子等小分子物質(zhì)的合成與存儲(chǔ)。而肝主疏泄生理功能與神經(jīng)系統(tǒng)對(duì)下丘腦-垂體-腎上腺(HPA)軸及其他機(jī)體反應(yīng)軸和神經(jīng)遞質(zhì)的調(diào)節(jié)有關(guān),也參與了 NEI網(wǎng)絡(luò)的調(diào)控機(jī)制。據(jù)此可得,血虛肝郁證證候本質(zhì)應(yīng)與NEI的整體調(diào)控機(jī)制具有高度相似性。因此,以血虛肝郁證動(dòng)物模型中NEI網(wǎng)絡(luò)生理和病理變化為研究對(duì)象,可初步闡明血虛肝郁證的證候的實(shí)質(zhì)。白芍,味苦、酸,性微寒,歸肝經(jīng)、脾經(jīng),即可"養(yǎng)血斂陰"為中醫(yī)傳統(tǒng)補(bǔ)血藥,又可"柔肝緩急"主治肝失調(diào)達(dá)、氣郁不舒。肝氣多郁,養(yǎng)血柔肝解郁是白芍臨床應(yīng)用的一個(gè)重要特色。白芍既能滋肝陰以養(yǎng)肝血,又能斂肝陰來柔肝體,最順應(yīng)肝之生理特性,為養(yǎng)血柔肝的代表性中藥。白芍,主入肝、脾經(jīng)。肝藏血主疏泄,主要體現(xiàn)在對(duì)氣機(jī)運(yùn)行和血液循行的調(diào)控上。其作用機(jī)制與現(xiàn)代醫(yī)學(xué)中內(nèi)分泌、免疫、循環(huán)、神經(jīng)、消化及生殖系統(tǒng)有關(guān)。脾為后天之本,主運(yùn)化水谷和水液,其生理功能與現(xiàn)代醫(yī)學(xué)消化、泌尿系統(tǒng)相關(guān);主生血、統(tǒng)血,提示其與內(nèi)分泌、循環(huán)系統(tǒng)相關(guān)。據(jù)此,可認(rèn)為白芍的功效與NEI網(wǎng)絡(luò)密切相關(guān)。芍藥苷、芍藥內(nèi)酯苷為白芍主要有效成分,芍藥內(nèi)酯苷為白芍特征性成分�，F(xiàn)代藥理研究表明,芍藥苷作為白芍的單萜類成分中含量最高的主要有效成分,大量實(shí)驗(yàn)證明芍藥苷主要作用于血液系統(tǒng)、神經(jīng)系統(tǒng)、內(nèi)分泌系統(tǒng),有抑制凝血酶原的生成、抗血小板聚集、擴(kuò)張靜脈及外周小血管、改善血液微循環(huán)、抗氧化、抗驚厥等多種生物學(xué)效應(yīng),提示其有補(bǔ)血、抗抑郁、鎮(zhèn)痛解痙和改善學(xué)習(xí)記憶以及對(duì)心腦血管的保護(hù)等作用。芍藥內(nèi)酯苷與芍藥苷化學(xué)結(jié)構(gòu)相似,是芍藥苷的同分異構(gòu)體;因其含量較芍藥苷低且難以分離純化,故藥理作用研究較少。本課題組基于自有專利技術(shù),對(duì)芍藥內(nèi)酯苷現(xiàn)有藥理作用研究結(jié)果表明,芍藥內(nèi)酯苷可作用于免疫系統(tǒng)中脾臟、胸腺和血液系統(tǒng)中造血細(xì)胞因子具有明確的補(bǔ)血作用,同時(shí)還可作用于神經(jīng)系統(tǒng)中HPA軸和腦內(nèi)單胺類神經(jīng)遞,具有明顯的抗抑郁作用。目的"證-效"相關(guān)理論提示中醫(yī)證候與中藥功效密切相關(guān),本研究以白芍"養(yǎng)血柔肝"功效為依據(jù),以血虛肝郁證動(dòng)物模型為切入點(diǎn),在系統(tǒng)梳理血虛肝郁證證候特點(diǎn)及芍藥苷、芍藥內(nèi)酯苷相關(guān)藥理作用研究的基礎(chǔ)上,觀察芍藥苷、芍藥內(nèi)酯苷對(duì)血虛肝郁證動(dòng)物模型大鼠的干預(yù)作用,通過檢測(cè)相關(guān)生理生化指標(biāo)以期從整體、組織、細(xì)胞和蛋白水平,探討白芍"養(yǎng)血柔肝"功效物質(zhì)基礎(chǔ)及作用機(jī)制和血虛肝郁證的證候?qū)嵸|(zhì)。方法本研究通過復(fù)制血虛肝郁證動(dòng)物模型和課題組自有白芍有效成分芍藥苷、芍藥內(nèi)酯苷分離純化專利技術(shù)的支撐下,以經(jīng)典補(bǔ)血方劑四物顆粒(含白芍且中國藥典以芍藥苷的含量作為質(zhì)量控制標(biāo)準(zhǔn))和傳統(tǒng)解郁方劑逍遙丸(含白芍且中國藥典以芍藥苷的含量作為質(zhì)量控制標(biāo)準(zhǔn))為陽性對(duì)照藥,并通過檢測(cè)各組大鼠體質(zhì)量,行為學(xué)實(shí)驗(yàn)(糖水消耗實(shí)驗(yàn)、敞箱實(shí)驗(yàn)、高架十字迷宮)以及外周血中WBC、RBC、HGB、脾臟指數(shù)、胸腺指數(shù)、腎上腺指數(shù)和血清中IL-3、IL-6、GM-CSF、TNF-α的含量和血漿中ACTH、CORT和下丘腦中CRH的水平、大腦皮質(zhì)中PKA、cAMP的活性、海馬組織中NE、DA、5-HT、E、5-HIAA的含量及cAMP/PKA信號(hào)通路相關(guān)神經(jīng)蛋白PKA、p-CREB、BDNF的蛋白表達(dá)水平的變化。結(jié)果1文獻(xiàn)研究1.1血虛肝郁證的辯證論治及其與神經(jīng)-內(nèi)分泌-免疫網(wǎng)絡(luò)關(guān)系的探討通過閱讀大量文獻(xiàn),較系統(tǒng)的梳理了中醫(yī)學(xué)對(duì)血虛肝郁證的辯證論治以及神經(jīng)-內(nèi)分泌-免疫網(wǎng)絡(luò)學(xué)說與中醫(yī)學(xué)相關(guān)理論的關(guān)系。不僅掌握了中醫(yī)學(xué)和現(xiàn)代醫(yī)學(xué)對(duì)血虛肝郁證的認(rèn)識(shí),而且明確了 NEI網(wǎng)絡(luò)與血虛肝郁證的相互關(guān)系,為血虛肝郁證的證候?qū)嵸|(zhì)研究提供了前期工作準(zhǔn)備。1.2芍藥苷、芍藥內(nèi)酯苷對(duì)神經(jīng)內(nèi)分泌免疫網(wǎng)絡(luò)藥理作用研究進(jìn)展通過閱讀大量文獻(xiàn),分別概括了白芍主要有效成分芍藥苷、芍藥內(nèi)酯苷對(duì)神經(jīng)系統(tǒng)、內(nèi)分泌系統(tǒng)、免疫系統(tǒng)藥理作用,揭示了白芍多環(huán)節(jié)、多靶點(diǎn)的藥理作用機(jī)制與神經(jīng)-內(nèi)分泌-免疫網(wǎng)絡(luò)密切相關(guān),即可為整體上系統(tǒng)研究白芍的傳統(tǒng)功效提供研究思路,也可為血虛肝郁證和白芍養(yǎng)血柔肝功效的"證-效"相關(guān)性研究提供借鑒意義。2實(shí)驗(yàn)研究2.1芍藥苷、芍藥內(nèi)酯苷對(duì)血虛肝郁證大鼠外周血細(xì)胞、臟器指數(shù)及造血細(xì)胞因子的影響結(jié)果:與模型組比較,PF30mg·kg-1組和AF30mg·kg-1組體質(zhì)量、脾臟指數(shù)、白細(xì)胞數(shù)量增加(P0.05、P0.01)。RIA結(jié)果顯示,PF30mg·kg-1和AF30mg·kg-1組均可增加該組 IL-3 含量(P0.05、P0.05)和減少該組 TNF-α 含量(P0.05、P0.05)。2.2芍藥苷、芍藥內(nèi)酯苷對(duì)血虛肝郁證大鼠行為學(xué)、HPA軸和單胺類神經(jīng)遞質(zhì)的影響結(jié)果:與模型組比較,PF30mg·kg-1組和AF30mg·kg-1組體質(zhì)量、糖水消量、曠場(chǎng)實(shí)驗(yàn)穿行次數(shù)增加(P0.05、P0.01)而滯留時(shí)間減少(P0.05、P0.05),高架十字迷宮實(shí)驗(yàn)開放臂進(jìn)入次數(shù)和停留時(shí)間明顯增加(P0.05、P0.05),而封閉臂停留時(shí)間減少(P0.05);RIA 結(jié)果顯示,PF30mg·kg-1 組和 AF30mg·kg-1 組均可降低該組 CORT、ACTH含量(P0.05、P0.05);HPLC-ECD 結(jié)果顯示,AF30mg·kg-1 組、PF30mg·kg-1 組均可不同程度的升高NE、DA、5-HA的含量(P0.05、P0.01)。2.3芍藥苷、芍藥內(nèi)酯苷對(duì)血虛肝郁證大鼠大腦皮層和海馬中cAMP、PKA含量及PKA、p-CREB、BDNF的蛋白水平的影響結(jié)果:與模型組比較,RIA結(jié)果顯示,PF30mg·kg-1組和AF30mg·kg-1組可使PKA含量升高,且AF30mg·kg-1組還可升高cAMP含量(P0.05、P0.05)。Western Blot結(jié)果顯示,與模型組比較,PF30mg·kg-1組和AF30mg·kg-1組均可增加該組PKA蛋白水平含量(P0.01、P0.05)。結(jié)論結(jié)論1:實(shí)驗(yàn)一結(jié)果表明,芍藥苷和芍藥內(nèi)酯苷作為白芍的主要有效成分,可通過對(duì)造血細(xì)胞因子的調(diào)控作用,調(diào)節(jié)免疫系統(tǒng)的紊亂,維持骨髓造血內(nèi)環(huán)境的穩(wěn)態(tài)。結(jié)論2:實(shí)驗(yàn)二結(jié)果表明,芍藥苷、芍藥內(nèi)酯苷作為白芍主要有效成分均可在一定程度上改善血虛肝郁大鼠行為學(xué)改變,其改善機(jī)制可能與二者調(diào)節(jié)神經(jīng)遞質(zhì)、減少應(yīng)激激素的分泌、抑制HPA軸的過度激活,調(diào)控神經(jīng)-內(nèi)分泌系統(tǒng)的功能有關(guān)。結(jié)論3:實(shí)驗(yàn)三結(jié)果表明,芍藥苷、芍藥內(nèi)酯苷可調(diào)控血虛肝郁證大鼠細(xì)胞內(nèi)cAMP/PKA信號(hào)通路中相關(guān)信號(hào)蛋白,發(fā)揮神經(jīng)保護(hù)作用。提示二者均為白芍養(yǎng)血柔肝功效的物質(zhì)基礎(chǔ)。綜上所述:本研究揭示白芍主要有效成分芍藥苷、芍藥內(nèi)酯苷可通過調(diào)節(jié)神經(jīng)-內(nèi)分泌-免疫(NEI)網(wǎng)絡(luò)對(duì)放射線輻照結(jié)合慢性束縛致血虛肝郁證動(dòng)物模型發(fā)揮養(yǎng)血柔肝功效,且提示中藥白芍與中醫(yī)血虛肝郁證的"證-效"相關(guān)性及血虛肝郁證的證候本質(zhì)的探討,可以神經(jīng)、內(nèi)分泌、免疫系統(tǒng)中相關(guān)細(xì)胞因子、激素、遞質(zhì)為突破口進(jìn)行NEI網(wǎng)絡(luò)的研究。并在此基礎(chǔ)上,本研究通過觀察芍藥苷、芍藥內(nèi)酯苷對(duì)血虛肝郁證動(dòng)物模型胞內(nèi)cAMP/PKA信號(hào)通路的影響,進(jìn)一步揭示白芍養(yǎng)血柔肝功效的作用機(jī)制還與對(duì)該信號(hào)通路中相關(guān)受體及靶點(diǎn)的調(diào)節(jié)作用有關(guān)。本研究不僅揭示了白芍"養(yǎng)血柔肝"功效的科學(xué)內(nèi)涵,探討了白芍主要有效成分芍藥苷、芍藥內(nèi)酯苷的藥理作用及機(jī)制,為白芍臨床應(yīng)用提供了科學(xué)依據(jù),而且通過血虛肝郁證和白芍養(yǎng)血柔肝功效"證-效"相關(guān)性的研究,既豐富了中醫(yī)肝臟證候的基礎(chǔ)研究,又可為中醫(yī)藥相關(guān)理論的研究提供借鑒意義。
[Abstract]:The syndrome of stagnation of the liver in the background of blood deficiency is the transfer of liver loss and relief caused by deficiency of liver blood deficiency, which not only has the deficiency of the liver and blood, but also the disorder of the liver Qi machine. It is a compound TCM syndrome of the coexistence of liver blood deficiency and liver loss and drainage. The function of liver and liver is closely related to the role of the liver in the metabolism of substance and energy in modern medicine, and can be involved in the liver. The synthesis and storage of peptide hormones and immune cell factors in the neural endocrine immune (NEI) network are synthesized and stored. The physiological function of liver catharsis is related to the regulation of the hypothalamus pituitary adrenal (HPA) axis and other body reaction axis and neurotransmitters, and also participates in the regulation mechanism of the NEI network. The essence of the syndrome of Qi syndrome should be highly similar to the overall regulation mechanism of NEI. Therefore, the physiological and pathological changes of NEI network in the animal model of blood deficiency liver depression are studied, and the essence of the syndrome can be clarified preliminarily. The main characteristic of the clinical application of Paeonia lactiflora is Paeonia lactiflora, which can not only nourish liver yin to nourish liver blood, but also converge liver yin to soft liver. It is the most conforming to the physiological characteristics of the liver. It is the representative traditional Chinese medicine for raising blood and liver. The mechanism is related to the endocrine, immune, circulation, nerve, digestion and reproductive systems in modern medicine. The spleen is the day after the spleen, the main transport of water and water, its physiological function with the modern medicine digestion, urinary system related; the main blood, the blood, suggesting that it is related to endocrine, circulation system. It is considered that the efficacy of Paeonia lactiflora is closely related to the NEI network. Paeoniflorin, paeoniflorin is the main effective component of Paeonia lactiflora, and paeoniflorin is a characteristic component of Paeonia lactiflora. Modern pharmacological studies have shown that paeoniflorin is the main effective component in the monoterpenoids of Paeonia lactiflora. A large number of experiments have proved that paeoniflorin mainly acts on blood. System, nervous system, endocrine system, inhibiting the generation of prothrombin, anti platelet aggregation, dilated veins and peripheral blood vessels, improving blood microcirculation, antioxidation, anticonvulsant and other biological effects, suggesting that it has the effect of blood supplement, antidepressant, analgesic and antispasmodic and improvement of learning and memory as well as the protection of heart and brain blood vessels. The chemical structure of glucoside and paeoniflorin is similar to the isomer of paeoniflorin. Because its content is lower than paeoniflorin and it is difficult to be separated and purified, the pharmacological action of paeoniflorin is less. Based on the patent technology, the research group has studied the pharmacological effects of paeoniflorin on the spleen, thymus and blood in the immune system. The hematopoietic cell factor in the system has a clear blood supplement and can also act on the HPA axis of the nervous system and the monoamine neurotransmitters in the brain and has an obvious antidepressant effect. On the basis of a systematic review of the characteristics of the syndrome of stagnation of the liver stagnation and the related pharmacological effects of paeoniflorin and paeoniflorin, the intervention effect of paeoniflorin and paeoniflorin on the rat model of liver depression with blood deficiency syndrome was observed, and the level of the whole, tissue, cell and protein level was discussed by detecting the related physiological and biochemical markers. Radix Paeoniae Paeoniae "nourishing blood and suppressing liver" effect material basis and function mechanism and syndrome essence of blood deficiency liver depression syndrome. Methods the study was carried out by replicating blood deficiency liver depression animal model and subjects group own Paeoniae paeoniflorin and paeoniflorin isolation and purification patent technology supported by the Chinese Pharmacopoeia (including Paeonia lactiflora and Chinese Pharmacopoeia). Taking the content of paeoniflorin as the quality control standard) and the traditional Jieyu prescription, Xiaoyao Pill (including Paeonia lactiflora and the content of paeoniflorin in Chinese Pharmacopoeia as the quality control standard) as the positive control drug, and by testing the body mass of rats in each group, behavior test (sugar water consumption experiment, open box experiment, elevated cross maze) and WBC, RBC, HGB in peripheral blood The contents of spleen index, thymus index, adrenal index, adrenal index and serum IL-3, IL-6, GM-CSF, TNF- alpha, the level of ACTH, CORT and CRH in the hypothalamus, the activity of PKA, cAMP in the cerebral cortex, the content of NE, DA, 5-HT, and related neuroproteins in the hippocampus, and the changes in the protein expression level. Results 1 literature study on the dialectical treatment of 1.1 blood deficiency syndrome of liver depression and its relationship with the neural endocrine immune network, through reading a large number of documents, systematically combed the dialectical treatment of blood deficiency syndrome in TCM and the relationship between the theory of neural endocrine immune network and the related theories of traditional Chinese Medicine. The mutual relationship between the NEI network and the syndrome of blood deficiency liver depression was clarified, and the relationship between the syndrome of blood deficiency and stagnation of liver depression was clarified, and the preliminary work for the study of syndrome essence of the syndrome of blood deficiency liver depression was prepared by.1.2 paeoniflorin, and the research progress on the pharmacological effects of paeoniflorin on the neuroendocrine immune network had passed through a large number of literature, and the main contents of Paeonia lactiflora were summarized. The pharmacological effects of paeoniflorin and paeoniflorin on the nervous system, the endocrine system and the immune system reveal the multiple links of Paeonia lactiflora. The pharmacological mechanism of the multiple targets is closely related to the neural endocrine immune network. It can provide a research idea for the study of the traditional work effect of Paeonia lactiflora on the whole and the syndrome of blood deficiency and the stagnation of the liver and the Paeonia lactiflora. The study of "syndrome effect" related research on the efficacy of nourishing blood and suppressing liver to provide reference significance.2 experimental study 2.1 paeoniflorin, paeoniflorin, the effects of paeoniflorin on peripheral blood cells, organ index and hematopoietic cytokines in rats with blood deficiency liver depression: compared with the model group, the body mass of PF30mg kg-1 group and AF30mg kg-1 group, the spleen index, and the number of white blood cells increased (P0.05, P0). .01).RIA results showed that the group of PF30mg / kg-1 and AF30mg / kg-1 could increase the IL-3 content of the group (P0.05, P0.05) and reduce the TNF- alpha content of the group (P0.05, P0.05) paeoniflorin, the effect of paeoniflorin on the behavior, the axis and the monoamine neurotransmitters of the blood deficiency liver depression rats: compared with the model group, the group and the group of minorities were compared with the model group. Body mass, sugar water dissipation, open field experiment transit times increased (P0.05, P0.01) and the retention time decreased (P0.05, P0.05). The entry times and residence time of the open arm labyrinth experiment increased significantly (P0.05, P0.05), while the retention time of the closed arm decreased (P0.05). RIA results showed that the PF30mg kg-1 group and AF30mg kg-1 group could reduce the group. RT, ACTH content (P0.05, P0.05); HPLC-ECD results showed that AF30mg kg-1 group, PF30mg. Kg-1 group can increase NE, DA, 5-HA content, paeoniflorin, the effect of paeoniflorin on the cerebral cortex and hippocampus of rats with blood deficiency liver depression The results of RIA showed that the content of PKA in the group of PF30mg / kg-1 and AF30mg. Kg-1 could increase the content of PKA, and the AF30mg / kg-1 group could also increase the cAMP content (P0.05, P0.05).Western. As the main effective component of paeoniflorin, glucoside and paeoniflorin can regulate the disturbance of the immune system and maintain the homeostasis of the hematopoietic environment in the bone marrow by regulating the hematopoietic cell factors. Conclusion the results of 2: experiment two show that paeoniflorin and paeoniflorin can improve blood deficiency liver depression to a certain extent as the main effective component of Paeonia lactiflora. Behavioral changes, its improvement mechanism may regulate neurotransmitters, reduce the secretion of stress hormones, inhibit the overactivation of HPA axis and regulate the function of the nervous endocrine system. Conclusion 3: experiment three showed that paeoniflorin and paeoniflorin could regulate the related signal eggs in the intracellular cAMP/PKA signaling pathway of blood deficiency liver depression rats. White Paeonia lactiflora is the material basis for the efficacy of Paeonia lactiflora to nourish blood and soften the liver. To sum up, this study reveals the main effective component of paeoniflorin, paeoniflorin, paeoniflorin, and paeoniflorin by regulating the neural endocrine immunization (NEI) network to nourish blood and soften the liver with radiation irradiation combined with chronic binding of blood deficiency syndrome. On the basis of the study of paeoniflorin and paeoniflorin on blood deficiency, the study on the correlation of "syndrome effect" and the essence of syndrome of stagnation of liver stagnation syndrome of Chinese traditional Chinese medicine and the syndrome essence of stagnation of liver Qi syndrome of blood deficiency can be suggested, and the related cytokines, hormones and transmitters in the immune system can be studied by NEI network. On this basis, the study was conducted by observing paeoniflorin and paeoniflorin in blood deficiency. The effect of the intracellular cAMP/PKA signaling pathway in the animal model of liver qi stagnation, further reveals that the mechanism of the effect of Paeonia lactiflora nourishing the blood and the liver is related to the regulation of the related receptors and targets in the signal pathway. This study not only revealed the scientific connotation of the efficacy of "nourishing the blood and liver" of Paeonia lactiflora, and discussed the main effective components of Paeonia Paeoniae paeoniflorin and Paeonia lactiflora. The pharmacological action and mechanism of ester glycosides provide scientific basis for the clinical application of Paeonia lactiflora, and the research on the correlation of "syndrome effect" through the syndrome of blood deficiency and stagnation of liver and the efficacy of Paeonia lactiflora, not only enriches the basic research of liver syndrome in Chinese medicine, but also provides reference for the research of TCM related theories.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285.5

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