慢性不可预知性应激与完全弗氏佐剂、福尔马林诱导的妊娠期疼痛-抑郁共病小鼠模型的构建与评价  

Establishment and Evaluation of Mouse Model of Pregnancy Pain-depression Comorbidity Induced by Chronic Unpredictable Stress,Complete Freund's Adjuvant and Formalin

作　　者：张一粟 刘欣茹 武若杰 刘睿 欧阳红 李晓红 ZHANG Yisu;LIU Xinru;WU Ruojie;LIU Rui;OUYANG Hong;LI Xiaohong(Department of Anesthesiology,The First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China;Graduate School of Bengbu Medical University,Bengbu 233000,China)

机构地区：[1]蚌埠医科大学第一附属医院麻醉科,蚌埠233000 [2]蚌埠医科大学研究生院,蚌埠233000

出　　处：《实验动物与比较医学》2024年第3期259-269,共11页Laboratory Animal and Comparative Medicine

基　　金：蚌埠医科大学研究生科研创新计划项目“基于海马区BDNF表达和GSK-3磷酸化交互作用探讨艾司氯胺酮对小鼠围生期疼痛-抑郁共病的作用和机制”(Byycx22098)

摘　　要：目的通过慢性不可预知性应激(chronic unpredictable stress,CUS)联合完全弗氏佐剂(complete Freund's adjuvant,CFA)和福尔马林诱导建立妊娠期疼痛-抑郁共病小鼠动物模型,对其相关表型进行系统评价,并初步探究其共病的病理基础。方法选择8周龄的C57BL/6J雌性小鼠,依据糖水偏好实验数据,采用随机分层方法将其分为对照组(妊娠前不干预)和CUS组(妊娠前予以CUS干预)。CUS造模完成后,将雌雄小鼠合笼配对,妊娠期小鼠分别于右后肢足底皮下注射50%CFA和5%福尔马林溶液,诱导建立妊娠期疼痛-抑郁共病小鼠模型。实验共分8组,即为对照-空白组、CUS-空白组、对照-CFA组、CUS-CFA组、对照-福尔马林组、CUS-福尔马林组、对照-CFA+福尔马林组、CUS-CFA+福尔马林组,每组10只。各组小鼠均在CUS干预前后、妊娠后和分娩后,通过糖水偏好实验、强迫游泳实验、悬尾实验和旷场实验评价行为学变化,并通过机械刺痛实验和热辐射痛实验测定各组小鼠对痛觉的敏感性变化;最后处死小鼠,用酶联免疫吸附法检测各组小鼠海马体中白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α),以及血清皮质醇(cortisol)和促肾上腺皮质激素(adrenocorticotropic hormone,ACTH)的含量。结果与对照-空白组相比,CUS-空白组模型小鼠产生了明显的抑郁样行为,并伴随痛阈明显降低(P<0.001)。与对照-空白组相比,对照-CFA+福尔马林组在CFA注射与福尔马林注射后均出现痛阈下降(P<0.01)。与对照-空白组和对照-福尔马林组相比,CUS-福尔马林组的痛阈明显降低(P<0.01),且三者依次下降。与对照-空白组和对照-CFA组相比,CUS-CFA组的痛阈明显降低(P<0.001),且三者依次下降。与对照-空白组、对照-CFA+福尔马林组相比,CUS-CFA+福尔马林组小鼠的机械痛阈显著降低(P<0.001)且三者依次下降,热辐射耐受时间缩短(P<0.01)且三者依次缩短。与Objective To establish a mouse model of pregnancy pain-depression comorbidity induced by chronic unpredictable stress(CUS),complete Freund's adjuvant(CFA),and formalin,and to systematically evaluate the associated phenotypes and preliminarily explore the pathological basis of the comorbidity.Methods Eight-week-old C57BL/6J female mice were randomly strarified divided into a control group(no intervention before pregnancy)and a CUS model group(CUS intervention before pregnancy)based on sucrose preference test(SPT)data.After completing the CUS treatment,female and male mice were paired and mated.Pain was induced by injecting 50%CFA and 5%formalin in the right hind foot during pregnancy to create a model of pregnancy pain-depression comorbidity.The experiment was divided into 8 subgroups:control-blank group,CUS-blank group,control-CFA group,CUS-CFA group,control-formalin group,CUS-formalin group,control-CFA+formalin group,and CUS-CFA+formalin group,with 10 mice in each group.The mice in each group were subject to behavioral tests,including the SPT,forced swimming test,tail suspension test,and open field test before and after CUS intervention,during pregnancy,and after delivery.Pain sensitivity changes were measured using mechanical allodynia and thermal hyperalgesia tests.Mice were then euthanized.Levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in hippocampus,as well as cortisol and adrenocorticotropic hormone(ACTH)in serum,were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the control-blank group,the CUS-blank group showed a significant depression-like behavior with reduced pain threshold(P<0.001).The control-CFA+formalin group showed a decrease in pain threshold after both CFA injection and formalin injection(P<0.01).Compared with the control-blank and control-formalin groups,the pain threshold was significantly lower in the CUS-formalin group(P<0.01),with a sequential decrease among the three.Compared with the control-blank and control-CFA groups,the pain threshold was

关 键 词：妊娠期抑郁 疼痛 共病 炎症因子 下丘脑-垂体-肾上腺轴 C57BL/6J小鼠 

分 类 号：R-332[医药卫生] Q95-33[生物学—动物学]