摘要
目的:通过整合药理学方法研究补肾安胎法代表方寿胎丸、健脾安胎法代表方举元煎调控自然流产(SA)蜕膜病理环节靶点逆转妊娠结局的安胎机制差异,初步阐释“胞络系肾”“胎茎系脾”生殖理论科学内涵。方法:运用GeneCards数据库筛选SA的调控靶点进行基因本体-生物学过程(GO-BP)富集分析,结合Cytoscape网络分析、文献引证筛选主要病理环节及核心靶点;利用高效液相色谱-质谱联用(HPLC-MS)筛选并获取寿胎丸、举元煎的潜在活性成分,构建其调控差异网络预测;复制SA大鼠模型进行同单位寿胎丸、举元煎干预,采用苏木素-伊红(HE)染色、透射电镜(TEM)、酶联免疫吸附测定法(ELISA)、免疫组化(IHC)、免疫荧光(IF)等方法进行安胎病理调控机制验证。结果:细胞黏附、炎症反应、细胞死亡、血管生成的调控失常是SA核心病理环节;分别获取寿胎丸潜在特异活性成分13个及举元煎活性成分14个,预测调控网络提示寿胎丸与举元煎的潜在活性成分均能调控血管内皮生长因子(VEGF)、白细胞介素(IL)-2、雌激素受体(ESR)-1、基质金属蛋白酶(MMP)-9等靶点从而调控SA病理环节;实验研究提示寿胎丸和举元煎可显著提高SA大鼠妊娠率、蜕膜细胞完整率及血供、有效控制细胞凋亡形态及雌激素(E2)、孕激素(P)及其受体表达,下调SA大鼠蜕膜组织MMP-2、MMP-9、IL-2、IL-6表达,同时上调抗凋亡蛋白B细胞淋巴瘤-2(Bcl-2)及IL-4表达。寿胎丸可显著上调VEGF表达,举元煎可显著下调E-钙黏蛋白(E-Cad)表达。结论:寿胎丸、举元煎均可通过调控SA核心病理过程以安胎,同条件干预下寿胎丸总体疗效优于举元煎,寿胎丸在调控血管生成途径优于举元煎,举元煎在细胞黏附途径优于寿胎丸,该结论可部分阐释“同病异治”生物学基础,为安胎中药质量标志物(Q-marker)识别、进一步方-证代谢组研究提供客观数据参考。
Objective:To explore difference in the mechanism of Shoutaiwan,a representative kidneytonifying and abortion-preventing formula,and Juyuanjian,a typical spleen-invigorating and abortionpreventing formula in reversing the pathology of decidua of spontaneous abortion(SA)patients and to expound the connotation of"uterine collaterals connecting kidney"and"fetal collaterals connecting spleen"theory.Method:The targets of SA were retrieved from GeneCards,followed by gene ontology-biological process(GO-BP)annotation.Based on Cytoscape and previous research,the main processes and core targets were screened out.High-performance liquid chromatography-mass spectrometry(HPLC-MS)was used to identify the potential active components of Shoutaiwan and Juyuanjian and the regulatory networks were constructed.SA was induced in rats and the model rats were treated with Shoutaiwan and Juyuanjian at the same unit.Hematoxylin and eosin(HE)staining,transmission electron microscopy(TEM),enzyme-linked immunosorbent assay(ELISA),immunohistochemistry(IHC),immunofluorescence(IF),and other methods were employed to verify the mechanisms against miscarriage.Result:The dysregulation of cell adhesion,inflammatory response,cell death,and angiogenesis was the core pathological process of SA.A total of 13 potential specific active components of Shoutaiwan and 14 active components of Juyuanjian were screened out.The regulatory networks showed that the potential active components of the two prescriptions modulated vascular endothelial growth factor(VEGF),interleukin(IL)-2,estrogen receptor(ESR)-1,matrix metalloproteinase-9(MMP-9),and other targets to regulate the pathological process of SA.The two can significantly improve the pregnancy rate and the integrity rate and blood supply of decidua cells,control the apoptosis morphology and the expression of estrogen(E2),progesterone(P),and its receptor,and down-regulate the expression of MMP-2,MMP-9,IL-2,and IL-6 in decidua tissue of SA rats.At the same time,they up-regulated the expression of anti-apoptotic protein B-cell lymphoma-2(Bcl-2)and IL-4.Shoutaiwan significantly up-regulated the expression of VEGF,and Juyuanjian significantly down-regulated the expression of E-cadherin(E-Cad).Conclusion:Both Shoutaiwan and Juyuanjian regulate the core pathological process of SA to prevent miscarriage.At the same unit,Shoutaiwan is overall superior to Juyuanjian.Shoutaiwan is better than Juyuanjian in regulating angiogenesis and Juyuanjian is superior to Shoutaiwan in regulating cell adhesion.This conclusion can partly explain the biological basis of"treating the same disease with different methods",and provide objective data reference for the identification of quality marker(Q-marker)of anti-miscarriage Chinese medicine and further study of formulasyndrome metabolome.
作者
周航
郑小艳
王欢
曾倩
邓琳雯
丁维俊
ZHOU Hang;ZHENG Xiaoyan;WANG Huan;ZENG Qian;DENG Linwen;DING Weijun(School of Basic Medicine,Chengdu University of Traditional Chinese Medicine(TCM),Chengdu 611137,China;Acupuncture and Tuina School,Chengdu University of TCM,Chengdu 611137,China;Sichuan College of TCM,Mianyang 621000,China;Hospital of Chengdu University of TCM,Chengdu 610075,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第20期186-200,共15页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81973901)
中国博士后科学基金面上项目(2021MD703798)
四川省科技厅计划项目(2021YJ0257)
成都中医药大学“杏林学者”学科人才科研提升计划项目(博士后专项)(0300058074)。
关键词
寿胎丸
举元煎
生物信息学分析
蜕膜
自然流产
差异机制
Shoutaiwan
Juyuanjian
bioinformatics analysis
decidua
spontaneous abortion
different mechanisms
