玄胡索散成分分析及其治療骨關(guān)節(jié)炎作用機制的網(wǎng)絡(luò)藥理學(xué)研究
發(fā)布時間：2019-05-29 21:36
【摘要】：骨關(guān)節(jié)炎是以進行性關(guān)節(jié)軟骨損傷、軟骨基質(zhì)合成與降解失衡造成關(guān)節(jié)面完全被破壞為特征的慢性關(guān)節(jié)病。目前主要采用保守治療、藥物治療和手術(shù)治療方式,通過延緩疾病發(fā)展、預(yù)防關(guān)節(jié)損傷從而達到緩解病癥的目的�；颊叨喾梅晴摅w抗炎藥、軟骨保護劑等化藥,但長期使用效果不佳,多有胃腸道反應(yīng)等毒副作用。中醫(yī)理論認(rèn)為骨關(guān)節(jié)炎屬"痹癥",病因為氣血痹阻、筋骨失養(yǎng),治療方法以補腎活血為主,輔以祛濕止痛。《普濟方》中收載的玄胡索散,由延胡索、牛膝、當(dāng)歸、補骨脂組成,用于治療肝腎不足,血凝氣滯,腰腿疼痛等。雖然只有四味藥,但組方精煉,體現(xiàn)了中醫(yī)治療骨關(guān)節(jié)炎的整體觀念及遣方用藥的配伍精髓。玄胡索散作為基礎(chǔ)方在臨床上應(yīng)用廣泛,并收載于以骨科聞名的中國中醫(yī)科學(xué)院望京醫(yī)院《骨科方劑藥方管理系統(tǒng)》中,但目前尚無針對該復(fù)方的相關(guān)研究報道,其治療骨關(guān)節(jié)炎的作用機制仍是空白,有必要進行深入研究,因此本試驗對玄胡索散進行了以下研究:1.玄胡索散及其拆方對補骨脂素與異補骨脂素溶出量的影響采用高效液相色譜法分析玄胡索散及其拆方對補骨脂素與異補骨脂素溶出量的影響,結(jié)果發(fā)現(xiàn)各拆方組較全方組有顯著差異(P0.05),其中當(dāng)歸、牛膝有助于二者的溶出,而延胡索抑制二者的溶出,為進一步揭示玄胡索散的配伍規(guī)律提供了依據(jù)。2.運用超高效液相色譜串聯(lián)四極桿飛行時間質(zhì)譜技術(shù)分析玄胡索散中所含化學(xué)成分基于超高效液相色譜串聯(lián)四極桿飛行時間質(zhì)譜技術(shù),首次對玄胡索散中所含主要化學(xué)成分進行了快速分離與定性,利用優(yōu)化后的分離與檢測條件在13 min內(nèi)對主要成分實現(xiàn)了有效分離,并通過與標(biāo)準(zhǔn)品、文獻及數(shù)據(jù)庫比較保留時間、碎片離子、裂解規(guī)律等信息,最終鑒定出57個化合物,為該藥后期的藥代動力學(xué)研究提供了依據(jù)。3.網(wǎng)絡(luò)藥理學(xué)預(yù)測玄胡索散治療骨關(guān)節(jié)炎的作用機制基于網(wǎng)絡(luò)藥理學(xué)方法,在相關(guān)數(shù)據(jù)庫及文獻檢索玄胡索散中四味中藥所含化學(xué)成分、化學(xué)成分靶點、骨關(guān)節(jié)炎治療靶點、蛋白相互作用關(guān)系等信息,并結(jié)合UPLC-Q-TOF-MS技術(shù)發(fā)現(xiàn)的配伍新成分,構(gòu)建相互作用網(wǎng)絡(luò),進行網(wǎng)絡(luò)藥理學(xué)預(yù)測。最終經(jīng)通路富集分析,發(fā)現(xiàn)了多條玄胡索散治療骨關(guān)節(jié)炎的信號通路,如調(diào)節(jié)NO、IL-1β的生物合成,調(diào)節(jié)炎癥反應(yīng)、響應(yīng)細胞因子或雌激素刺激,以及抗軟骨細胞凋亡等。本研究在系統(tǒng)層面首次揭示了該藥治療骨關(guān)節(jié)炎的作用機制,并得到了相關(guān)化學(xué)成分靶點、治療通路等之間的密切關(guān)系,為后期的試驗研究提供了依據(jù)。4.玄胡索散對骨關(guān)節(jié)炎大鼠軟骨基質(zhì)的影響雄性SD大鼠隨機分組,采用前交叉韌帶橫斷聯(lián)合部分半月板切除造骨關(guān)節(jié)炎模型,分別于給藥3周、6周后處死大鼠取手術(shù)側(cè)膝關(guān)節(jié),進行HE染色切片評估軟骨損傷程度與Masson染色切片評估膠原的降解。結(jié)果發(fā)現(xiàn)玄胡索散能明顯抑制膝關(guān)節(jié)軟骨損傷與膠原降解(P0.05),初步證明了玄胡索散治療骨關(guān)節(jié)炎的潛在作用,為后期的體外細胞試驗與網(wǎng)絡(luò)藥理學(xué)預(yù)測結(jié)果驗證試驗提供了依據(jù)。
[Abstract]:Osteoarthritis is a chronic arthropathy characterized by progressive joint cartilage injury, cartilage matrix synthesis and degradation imbalance resulting in the complete destruction of the articular surface. The invention mainly adopts the conservative treatment, the medicine treatment and the operation treatment mode, and the purpose of relieving the condition is achieved by delaying the development of the disease and preventing the joint injury. The patient has the advantages of no anti-inflammatory agent, chondroprotective agent and the like, but the long-term use effect is not good, and has the toxic and side effects such as gastrointestinal reaction and the like. The traditional Chinese medicine theory considers that the osteoarthritis belongs to the "a hand", and the disease is due to the obstruction of qi and blood, the loss of bones and bones, and the treatment method, which is mainly used for tonifying the kidney and promoting blood circulation, and is supplemented with the functions of eliminating And is prepared from Rhizoma Corydalis, Achyranthis Radix, Radix Angelicae Sinensis, and Fructus Psoraleae, and can be used for the treatment of deficiency of liver and kidney, blood coagulation and qi stagnation, and lumbago. Although only four medicines are used, the prescription is refined, and the integral concept of the traditional Chinese medicine for treating the osteoarthritis and the compatibility essence of the prescription drugs are reflected. It is widely used as a base in the clinical application, and is listed in the Wangjing Hospital of the Chinese Academy of Traditional Chinese Medicine, which is known as the Orthopaedics, and in the Prescription Management System of Orthopaedics. However, there is no relevant research report to the compound, and the mechanism for the treatment of osteoarthritis is still blank. It is necessary to carry out the in-depth study, so this test has carried out the following research on Xhosoxuan:1. The effect of Xuanhuoxue and its removal on the amount of psoralen and isopsoralen was analyzed by high performance liquid chromatography (HPLC). The results showed that there was a significant difference in the amount of psoralen and isopsoralen (P0.05). Achyranthis bidentata is helpful for the dissolution of the two, while the dissolution of the two can provide a basis for further revealing the compatibility of the Xuanhuoxue powder. The high performance liquid chromatography tandem quadrupole time-of-flight mass-spectrum technique is used to analyze the chemical components contained in the Xuanhuizu powder based on the ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry technology, and the first time the main chemical components contained in the Xhososol powder are rapidly separated and characterized, By using the optimized separation and detection conditions, the main components are effectively separated within 13 minutes, and the information such as retention time, fragment ion, cracking rule and the like is compared with the standard, the literature and the database, and the 57 compounds are finally identified, It provides a basis for the study of the pharmacokinetics of the drug in the later stage. The invention relates to a network pharmacology prediction function mechanism for treating osteoarthritis, which is based on the network pharmacology method, and the information such as the chemical components, the chemical components target point, the treatment target point of the osteoarthritis treatment, the protein interaction relation and the like contained in the four traditional Chinese medicines in the relevant database and the literature search, Combined with the new components of UPLC-Q-TOF-MS, the interaction network was constructed and the network pharmacological prediction was carried out. And finally, through the channel enrichment analysis, a plurality of signal paths for treating the osteoarthritis, such as regulating the biological synthesis of NO and IL-1, regulating the inflammatory reaction, responding to the cytokine or the estrogen stimulation, and the anti-chondrocyte apoptosis, and the like are found. In this study, the mechanism of the treatment of osteoarthritis was first revealed at the system level, and the close relationship between the relevant chemical components and the treatment path was obtained. The male SD rats were randomly divided into two groups: the anterior cruciate ligament transection and the joint part of the meniscectomy for osteoarthritis, and the knee joint of the operation side was sacrificed after 6 weeks of administration. The degree of cartilage damage was assessed by HE staining and the degradation of the collagen was assessed by the Masson staining section. The results showed that the treatment of the articular cartilage injury and the degradation of the collagen (P0.05). The potential role of the Xuanhuoxue powder in the treatment of osteoarthritis was preliminarily proved, and the basis was provided for the later in vitro cell test and the results of the validation of the network pharmacological prediction.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285
[Abstract]:Osteoarthritis is a chronic arthropathy characterized by progressive joint cartilage injury, cartilage matrix synthesis and degradation imbalance resulting in the complete destruction of the articular surface. The invention mainly adopts the conservative treatment, the medicine treatment and the operation treatment mode, and the purpose of relieving the condition is achieved by delaying the development of the disease and preventing the joint injury. The patient has the advantages of no anti-inflammatory agent, chondroprotective agent and the like, but the long-term use effect is not good, and has the toxic and side effects such as gastrointestinal reaction and the like. The traditional Chinese medicine theory considers that the osteoarthritis belongs to the "a hand", and the disease is due to the obstruction of qi and blood, the loss of bones and bones, and the treatment method, which is mainly used for tonifying the kidney and promoting blood circulation, and is supplemented with the functions of eliminating And is prepared from Rhizoma Corydalis, Achyranthis Radix, Radix Angelicae Sinensis, and Fructus Psoraleae, and can be used for the treatment of deficiency of liver and kidney, blood coagulation and qi stagnation, and lumbago. Although only four medicines are used, the prescription is refined, and the integral concept of the traditional Chinese medicine for treating the osteoarthritis and the compatibility essence of the prescription drugs are reflected. It is widely used as a base in the clinical application, and is listed in the Wangjing Hospital of the Chinese Academy of Traditional Chinese Medicine, which is known as the Orthopaedics, and in the Prescription Management System of Orthopaedics. However, there is no relevant research report to the compound, and the mechanism for the treatment of osteoarthritis is still blank. It is necessary to carry out the in-depth study, so this test has carried out the following research on Xhosoxuan:1. The effect of Xuanhuoxue and its removal on the amount of psoralen and isopsoralen was analyzed by high performance liquid chromatography (HPLC). The results showed that there was a significant difference in the amount of psoralen and isopsoralen (P0.05). Achyranthis bidentata is helpful for the dissolution of the two, while the dissolution of the two can provide a basis for further revealing the compatibility of the Xuanhuoxue powder. The high performance liquid chromatography tandem quadrupole time-of-flight mass-spectrum technique is used to analyze the chemical components contained in the Xuanhuizu powder based on the ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry technology, and the first time the main chemical components contained in the Xhososol powder are rapidly separated and characterized, By using the optimized separation and detection conditions, the main components are effectively separated within 13 minutes, and the information such as retention time, fragment ion, cracking rule and the like is compared with the standard, the literature and the database, and the 57 compounds are finally identified, It provides a basis for the study of the pharmacokinetics of the drug in the later stage. The invention relates to a network pharmacology prediction function mechanism for treating osteoarthritis, which is based on the network pharmacology method, and the information such as the chemical components, the chemical components target point, the treatment target point of the osteoarthritis treatment, the protein interaction relation and the like contained in the four traditional Chinese medicines in the relevant database and the literature search, Combined with the new components of UPLC-Q-TOF-MS, the interaction network was constructed and the network pharmacological prediction was carried out. And finally, through the channel enrichment analysis, a plurality of signal paths for treating the osteoarthritis, such as regulating the biological synthesis of NO and IL-1, regulating the inflammatory reaction, responding to the cytokine or the estrogen stimulation, and the anti-chondrocyte apoptosis, and the like are found. In this study, the mechanism of the treatment of osteoarthritis was first revealed at the system level, and the close relationship between the relevant chemical components and the treatment path was obtained. The male SD rats were randomly divided into two groups: the anterior cruciate ligament transection and the joint part of the meniscectomy for osteoarthritis, and the knee joint of the operation side was sacrificed after 6 weeks of administration. The degree of cartilage damage was assessed by HE staining and the degradation of the collagen was assessed by the Masson staining section. The results showed that the treatment of the articular cartilage injury and the degradation of the collagen (P0.05). The potential role of the Xuanhuoxue powder in the treatment of osteoarthritis was preliminarily proved, and the basis was provided for the later in vitro cell test and the results of the validation of the network pharmacological prediction.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285
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