【作者】 张寒；

【导师】 李得加；

【作者基本信息】 武汉大学 ， 劳动卫生与环境卫生学， 2021， 硕士

【摘要】 目的:绝经后骨质疏松(postmenopausal osteoporosis,PMOP)属于原发性骨质疏松,其较高发病率及其骨质疏松性骨折的并发症,已严重威胁中老年妇女人群的健康,成为当今社会主要的公共卫生问题。本研究旨在通过网络药理学和转录组测序技术探究补肾化痰方治疗PMOP的研究机制。方法:通过中药系统药理学(TCMSP)、中医大百科全书(ETCM)和中药分子机理的生物信息学分析工具(BATMAN-TCM)等网络药理学工具筛选出补肾化痰方中六味中药的有效生物活性成分,利用生物信息学分析治疗PMOP的潜在靶点的主要信号通路。建立假手术组(Sham)、绝经后骨质疏松模型组(OVX)和补肾化痰方中药组(OVX-MED)。基于转录组测序方法,对三组大鼠血液样本进行分析,进一步明确补肾化痰方对差异表达基因的调控作用。结果:通过数据库筛选获得补肾化痰方的生物活性成分54种,治疗PMOP作用靶点1107个,骨质疏松症相关疾病基因靶点944个;绘制维恩图并计算交集共得出80个共同靶点,通过与转录组测序获得的靶点交集,得到补肾化痰方干预PMOP的核心基因,包括肿瘤坏死因子(TNF)、核转录因子(JUN)、白细胞介素-6(IL-6)等;对核心基因进行分析,GO功能注释结果表明其生物学过程主要包括对药物反应、RNA聚合酶II启动子转录的正调控、凋亡过程的负调控和衰老等;分子功能主要包括酶结合、类固醇激素受体、类固醇结合、细胞因子活性等;细胞成分主要包括细胞外间隙、胞外区、小窝、细胞表面和细胞外基质等;而KEGG通路富集分析结果表明,补肾化痰方参与骨质疏松症相关的信号通路如Toll样受体信号通路、T细胞受体信号通路、TNF信号通路、破骨细胞分化和HIF-1信号通路等;补肾化痰方可以抑制炎症反应,调节机体骨免疫的微环境,进一步参与成骨细胞以及破骨细胞的分化、增殖、凋亡等生理过程,减少氧化应激造成的损伤,同时调控细胞因子。结论:补肾化痰方通过多靶点、多途径干预PMOP,发挥出中药复方治疗PMOP的优势。更多还原

【Abstract】 Objective Postmenopausal osteoporosis(PMOP)belongs to primary osteoporosis.Its high incidence and the complications of osteoporotic fractures have seriously threatened the health of middle-aged and elderly women,and become a major public health problem in today’s society.This study aims to explore the research mechanism of Bushen Huatan Recipe in the treatment of PMOP through network pharmacology and transcriptome sequencing technology.Methods Through network pharmacology tools such as TCM System Pharmacology(TCMSP),Encyclopedia of Traditional Chinese Medicine(ETCM)and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMANTCM)and other network pharmacological tools,the effective biologically active ingredients of six traditional Chinese medicines in Bushen Huatan Recipe were screened out.Using bioinformatics to analyze the main signal pathways of potential targets for the treatment of PMOP.The sham operation group(Sham),postmenopausal osteoporosis model group(OVX)and Chinese medicine group of Bushen Huatan Recipe(OVX-MED)were established.Based on the transcriptome sequencing method,three groups of rat blood samples were analyzed to further clarify the regulating effect of Bushen Huatan Recipe on differentially expressed genes.Results Through database screening,54 biologically active ingredients of Bushen Huatan Recipe,1107 targets for the treatment of PMOP,and 944 gene targets for osteoporosis-related diseases were obtained.Draw a Venn diagram and calculate the intersection to get a total of 80 common targets.Through the intersection with the targets obtained by transcriptome sequencing,the core genes of Bushen Huatan Recipe intervention in PMOP were obtained,including tumor necrosis factor(TNF),nuclear transcription factor(JUN),interleukin-6(IL-6)and so on.Analyzing the core genes,GO functional annotation results show that its biological process mainly includes drug response,positive regulation of RNA polymerase II promoter transcription,negative regulation of apoptosis process,and aging.The molecular functions mainly include enzyme binding,steroid hormone receptor,steroid binding,cytokine activity and so on.Cell components mainly include extracellular space,extracellular area,pits,cell surface and extracellular matrix.The results of enrichment analysis of KEGG pathway show that Bushen Huatan Recipe is involved in osteoporosis-related signaling pathways such as Toll-like receptor signaling pathway,T cell receptor signaling pathway,TNF signaling pathway,osteoclast differentiation and HIF-1 signaling.Bushen Huatan Recipe can inhibit inflammation,regulate the microenvironment of the body’s bone immunity,and further participate in the physiological processes of osteoblast and osteoclast differentiation,proliferation,and apoptosis,reduce damage caused by oxidative stress,and regulate cytokines.Conclusion Bushen Huatan Recipe intervenes PMOP through multiple targets and multiple channels,and exerts the advantages of traditional Chinese medicine compound prescriptions for PMOP treatment.更多还原