黄体酮撤退加情志刺激诱导型经前期综合征大鼠模型的建立与评价

Establishment and evaluation of a premenstrual syndrome rat model induced by progesterone-withdrawal and emotional-stimulation

经前期综合征(PMS)缺乏与中西医临床诊疗标准吻合度高、可行性好且表现稳定的动物模型,导致其发病机制、药效药理的相关研究缺乏可靠的实验载体。该研究旨在从“病-证-症”关联视角,系统解析PMS的生物内涵,在此基础上建立黄体酮撤退与情志刺激结合诱导型PMS动物模型,并对其进行客观评价。首先通过GeneCards、DisGeNET、MalaCards和中医证候本体及多维定量关联计算平台(SoFDA)数据库与已发表文献,收集PMS现代医学与中医诊疗标准中肝郁气滞与气滞血瘀证型相关的临床症状基因集,并依据基因间相互作用信息,建立PMS的“病-证-症”关联网络。基于数据挖掘结果,在经典黄体酮撤退模型基础上,结合慢性束缚刺激,模拟临床发病过程中由外部环境刺激引起的情绪抑郁,从生理与情志2个维度引起病理损伤,制备PMS大鼠模型,评价2种造模条件下的模型在旷场实验得分,脏器指数,卵巢病理变化,血清雌二醇(E_2)、促卵泡激素/促黄体生成素(FSH/LH)、5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)水平及凝血四项、血流变指标。通过计算“病-证-症”关联网络节点在网络中的度值、介度和接近度,筛选出163个具有网络拓扑重要性的核心基因。进一步的生物功能挖掘结果表明,PMS核心基因主要参与“神经-内分泌-免疫”系统与血液循环障碍相关通路的调控;临床表型症状基因集映射分析提示,PMS核心基因与抑郁症状及血瘀所致肿痛症状显著相关。与单纯的黄体酮撤退模型相比,注射与束缚双重刺激的大鼠在旷场实验水平得分与总分,卵巢组织病理变化,血清神经递质5-HT与DA水平,血清性激素E_2与FSH/LH水平的异常改变均更为显著,且注射加束缚条件加剧了模型大鼠血液流变性、凝血功能和红细胞形态的病理改变,证实了该大鼠模型能够表征PMS发生、进展中的“神经-内分泌-免疫”系统紊乱与血液循环系统障碍,与该疾病的中西医临床诊疗标准相符。黄体酮撤退与情志刺激结合诱导型PMS大鼠模型的建立能够为PMS发病机制的阐释、治疗药物的发现与药效评价提供可靠的实验载体,也为客观化反映PMS中西医临床表征与精准治疗提供参考。

Premenstrual syndrome(PMS)lacks a highly consistent and feasible animal model that aligns with diagnostic and therapeutic standards in both traditional Chinese medicine(TCM)and western medicine,resulting in a lack of reliable experimental carriers for studying its pathogenesis and pharmacological effects.This study aims to systematically analyze the biological implications of PMS from the perspective of the"disease-syndrome-symptom"correlation and establish preparation and evaluation methods for an improved animal model of this disease.Firstly,clinical symptom gene sets related to the Qi stagnation syndromes due to liver depression and blood stasis in PMS in both modern medicine and TCM diagnostic standards were collected through GeneCards,DisGeNET,Mala-Cards,and the System of Foundational Diagnostic Association(SoFDA)database,as well as published literature.Based on the interaction information between genes,a"disease-syndrome-symptom"correlation network of PMS was established.Based on data mining results,an improved rat model of PMS was prepared by combining chronic restraint stress with the classical progesterone-withdrawal mo-del to simulate emotional depression caused by external environmental stimuli during the clinical onset process,inducing pathological damage from both physiological and emotional dimensions.The evaluation of the improved model before and after modification included open field experiment scores,organ indices,ovarian pathological changes,serum levels of estradiol(E_2),follicle-stimulating hormone/luteinizing hormone(FSH/LH),5-hydroxytryptamine(5-HT),dopamine(DA),norepinephrine(NE),as well as coagulation parameters and hemorheology indexes.By calculating the degree,betweenness,and closeness centrality of nodes in the"disease-syndrome-symptom"correlation network,163 core genes with topological importance were identified.Further biological function mining results indicated that core genes in PMS mainly participated in the regulation of the"nervous-endocrine-immune"system and pathways related to circulatory disorders.Mapping analysis of clinical phenotype symptom gene sets suggested significant correlations between core genes in PMS and depressive symptoms and pain symptoms caused by blood stasis.Compared with the simple progesterone withdrawal model,rats subjected to combined injections and restraint stress showed more significant abnormalities in open field experiment scores,ovarian tissue pathology,serum neurotransmitter levels of 5-HT and DA,as well as serum hormone levels of E_2 and FSH/LH.The modified modeling conditions exacerbated the pathological changes in blood rheology,coagulation function,and red blood cell morphology in model rats,confirming that the improved rat model could characterize the"nervous-endocrine-immune"system disorder and circulatory system disorders in the occurrence and progression of PMS,consistent with the clinical diagnostic and therapeutic standards of both TCM and western medicine.The establishment of the improved rat model of PMS can provide a reliable experimental carrier for elucidating the pathogenesis of PMS and discovering and evaluating therapeutic drugs.It also provides references for objectively reflecting the clinical characteristics of PMS in TCM and western medicine and precision treatment.