浙派裘氏妇科治疗子宫内膜异位症的共性规律及常用药对分子机制的研究

Research on Common Law of Endometriosis Treatment of QIU’s Gynecology in Zhejiang Province and Molecular Mechanism of Common Drug Pair

[目的]通过对浙江省裘氏妇科流派治疗子宫内膜异位症(endometriosis,EMs)的中医思辨特征、共性规律的研究,总结裘氏妇科诊疗EMs的群体共性规律,并且基于网络药理学研究共性规律中的常用药对的分子机制,以期传承浙派裘氏妇科临证用药经验,提高临床疗效。[方法]临床随诊在省内具有一定影响力的4位裘氏妇科传承人张萍青、吴燕平、王幸儿、张婷,规范采集和录入临床诊疗信息,建立名中医医案数据库,采用聚类分析、关联规则分析等技术,对各名中医的用药特点、辨证规律等进行数据分析挖掘,总结其共性规律。通过中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)挖掘常用药对的活性成分及作用靶标,通过GeneCards数据库挖掘EMs的相关靶点,筛选出常用药对治疗EMs的靶标,利用Cytoscape软件构建活性成分-EMs靶标网络。利用生物学信息注释数据库进行基因本体(gene ontology,GO)生物过程与京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)代谢通路富集分析,预测常用药对治疗EMs的分子机制。[结果]EMs临床常见证型包括瘀热互结证、肾虚血瘀证、气滞血瘀证。针对不同证型,4位医家辨证论治,同中存异,治疗瘀热互结证均以苦寒类的清热药及活血化瘀药为主,治疗肾虚血瘀证使用最多的药物均为补虚药及活血化瘀药,针对气滞血瘀证使用最多的药物均为理气药及活血化瘀药。其中丹参-赤芍药对是裘氏妇科治疗EMs的常用药对,通过TCMSP数据库共筛选出该药对的67种活性成分及78个治疗EMs的靶标,丹参-赤芍药对通过影响信号转导和转录活化因子3(signal transducer and activator of transcription 3,STAT3)、蛋白激酶B1(protein kinase B1,PKB1)、转录因子激活蛋白-1(activator protein-1,AP-1/JUN)、丝裂原活化蛋白激酶1(mitogen activated protein kinase 1,MAPK1)、核转录因子-κB p65(nuclear factor-κB p65,NF-κB p65/RELA)等关键靶点,调控磷脂酰肌醇3-激酶-蛋白激酶B(phosphatidylinositol 3-kinase-protein kinases B,PI3K-PKB)信号通路、白细胞介素-17(interleukin-17,IL-17)信号通路、缺氧诱导因子-1(hypoxia inducible factor-1,HIF-1)信号通路等治疗EMs。[结论]4位裘氏妇科传承人治疗EMs,均以“肾虚”为本,“瘀”“热”互结为标,以苦寒、甘温之品补肾虚、化瘀血、清瘀热、行气滞,且善用药对。丹参-赤芍药对为裘氏妇科的常用药对,与裘笑梅先生清化逐瘀治疗EMs的学术思想相吻合。网络药理学揭示了该药对治疗EMs的分子机制,为进一步药效物质基础分析和分子机制研究提供了依据。

[Objective]By studying the speculative characteristics and common law of traditional Chinese medicine(TCM) in the treatment of endometriosis(EMs) of QIU’s Gynecology in Zhejiang Province, the common law of QIU’s Gynecology in diagnosis and treatment of EMs was summarized. In addition, based on network pharmacology, the molecular mechanism of common drugs pair in the treatment of EMs was studied in order to inherit the experience in clinical application of drugs of QIU’s Gynecology and improve the clinical efficacy.[Methods]Through clinicaly following up the four inheritors of QIU’s Gynecology who have a certain influence in the province, ZHANG Pingqing, WU Yanping, WANG Xing’er and ZHANG Ting, the collection and input of clinical diagnosis and treatment information was standardized, the database of famous TCM medical cases was established, and the cluster analysis, association rule analysis and other technologies were adopted to analyze and explore the data of medication characteristics and syndrome differentiation rules, and their common laws were summarized. The active ingredients and effect of targets of common drug pairs were explored by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) database;the relevant targets of EMs were explored by GeneCards database;the targets of common drug pair for treatment of EMs were screened out, and the active ingredient-EMs target network was constructed by Cytoscape software. By using the annotation database of biological information(DAVID), gene ontology(GO) biological process and Kyoto Encyclopedia of Genes and Genomes(KEGG) metabolic pathway enrichment analysis were conducted to predict the molecular mechanism of common drug pair for treatment of EMs. [Results]Mutual knot syndrame of blood stasis and heat, kidney deficiency and blood stasis, and Qi stagnation and blood stasis are common clinical syndromes of EMs. Four doctors treat different syndrome types according to syndrome differentiation, as well as seek common ground with reserving differences. In the treatment of the mutual knot syndrome of blood stasis and heat, the four doctors mainly used cold and bitter heat-clearing drugs and blood-activating drugs to remove blood stasis. In the treatment of kidney deficiency and blood stasis syndrome, the most common drugs used by the 4 doctors were deficiency tonifying and blood-activating drugs. While for the Qi stagnation and blood stasis syndrome, the most common drugs used by the 4 doctors were Qi regulating drugs and drugs for promoting blood circulation and removing blood stasis. Radix Salviae Miltiorrhizae-Radix Paeoniae Rubra is the common drug pair of QIU’s Gynecology for the treatment of EMs. A total of 67 active ingredients and 78 targets of EMs were screened out by TCMSP database. Radix Salviae Miltiorrhizae-Radix Paeoniae Rubra treat EMs by influencing key targets such as signal transducer and activator of transcription 3(STAT3), protein kinase B1(PKB1), activator protein-1(AP-1/JUN), mitogen activated protein kinase 1(MAPK1) and nuclear factor-κB p65(NF-κB p65/RELA) and regulating phosphatidylinositol 3-kinase-protein kinases B(PI3K-PKB) signaling pathway,interleukin-17(IL-17) signaling pathway and hypoxia inducible factor-1(HIF-1) signaling pathway.[Conclusion]The four QIU’s Gynecology inheritors all treat EMs with the “kidney deficiency” as the basis, and the “blood stasis” and “heat” as the manifestation. In the treatment of EMs, they use drugs of bitter and cold as well as sweet and warm to reinforce the kidney, remove blood stasis, clear away heat stasis and activate Qi stagnation, and good at using drug pair. As the common medicine pair of QIU’s Gynecology, Radix Salviae Miltiorrhizae-Radix Paeoniae Rubra is consistent with QIU Xiaomei's academic thought of EMs. The network pharmacology reveals the molecular mechanism of the drug pair in the treatment of EMS, and provides a basis for further analysis of the material basis and molecular mechanism of its efficacy.