非手术子宫注射法建立大鼠盆腔炎模型的研究  

Establishment of a rat model of pelvic inflammatory disease via non-operative uterine injection

作　　者：杨美艳 颜峰[3] 王雪楠[4] 胡凯歆 高鸿霞 潘晓燕[2] YANG Meiyan;YAN Feng;WANG Xuenan;HU Kaixin;GAO Hongxia;PAN Xiaoyan(School of Medical Technology,Beihua University,Jilin 132013,China;Center for Reproductive Medicine,Jilin Medical University,Jilin 132013;Qufu City People’s Hospital,Qufu 273100;Department of Reproduction,Affiliated Hospital of Jining Medical College,Jining 272007)

机构地区：[1]北华大学医学技术学院,吉林132013 [2]吉林医药学院生殖医学研究中心,吉林132013 [3]曲阜市人民医院,山东曲阜273100 [4]济宁医学院附属医院生殖科,山东济宁272007

出　　处：《中国比较医学杂志》2025年第1期13-29,共17页Chinese Journal of Comparative Medicine

基　　金：吉林省科技厅重点攻关项目(20240305074YY);吉林省发展和改革委员会产业技术研究与开发项目(2023C027-8);山东省中医药科技项目(Z-2023065);吉林医药学院研究生创新计划项目(2023yyc02)。

摘　　要：目的非手术法建立高效、稳定的大鼠盆腔炎模型,并评估验证其在药效试验中的应用。方法雌性SD大鼠随机分为对照组、苯酚7 d模型组、苯酚10 d模型组、苯酚造模治疗组、低浓度菌液模型组、高浓度菌液模型组和菌液造模治疗组。模型组和治疗组SD大鼠以非手术法子宫腔注射25%苯酚凝胶、2×10^(7)个或2×10^(8)个大肠埃希菌和金黄色葡萄链球菌的混合菌,以构建盆腔炎模型。治疗组大鼠灌胃中成药金刚藤胶囊,对照组大鼠在相同时间给予等体积的溶剂溶液。观察大鼠的健康状况、体重变化和子宫外观;计算子宫系数;苏木素-伊红(HE)染色检测子宫和输卵管的病理变化、子宫内膜厚度和腺体数量;酶联免疫吸附试验(ELISA)检测大鼠血清中白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor,TNF-α)水平;免疫荧光染色检测巨噬细胞标志物CD68蛋白表达;Western blot检测大鼠子宫中TLR4/NF-κB通路相关蛋白表达。结果模型组大鼠的死亡率只有5%;与对照组相比,造模后大鼠体重下降、子宫系数增加、子宫和输卵管出现不同程度的病理变化、内膜变薄、腺体数量减少、血清IL-1β、IL-6、TNF-α的水平和子宫组织中巨噬细胞的数量显著增加、Toll样受体4/核因子-κB(Toll-like receptor 4/nuclear factor kappa-B,TLR4/NF-κB)信号通路被激活;苯酚7 d模型和低浓度菌液模型被判定为轻度盆腔炎模型,苯酚10 d模型和高浓度菌液模型被判定为重度盆腔炎模型;中成药金刚藤胶囊治疗后,子宫和输卵管的病理症状明显缓解,符合临床盆腔炎的药效评估。结论通过非手术法利用苯酚和混合菌液建立的大鼠盆腔炎模型可以更好地模拟临床由于不同原因导致的不同程度病理状态的盆腔炎,适用于药物药效作用评价和盆腔炎病理机制的阐明。Objective To establish an efficient and stable model of pelvic inflammatory disease in rats via a non-surgical method,and to evaluate its application in pharmacodynamic testing.Methods Female Sprague-Dawley rats were divided randomly into the following groups:control group;model group with phenol for 7 d;model group with phenol for 10 d;treatment group modeled with phenol;model group with low concentration of bacteria;model group with high concentration of bacteria;and treatment group modeled with bacteria.Rats in the model and treatment groups were injected with 25%phenol gel and 2×10^(7) or 2×10^(8) Escherichia coli and Staphylococcus aureus mixture via a non-surgical method,to construct a rat model of pelvic inflammatory disease.Rats in the treatment groups received the Chinese patent medicine Jingangteng capsules by gavage,and rats in the control group received the same volume of solvent solution.The health status,weight changes,and uterine appearance were monitored and the uterine coefficient was calculated.Pathological changes in the uterus and fallopian tubes,endometrial thickness,and number of glands were detected by hematoxylin and eosin staining.Serum levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay.Protein expression of the macrophage marker CD68 was detected by immunofluorescence.Expression of Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB pathway-related proteins in the uterus was detected by Western blot.Results The mortality rate in the model group was only 5%.Compared with the control group,model rats showed decreased body weight,increased uterine coefficient,pathological changes in the uterus and Fallopian tubes,thinner endometrium,fewer glands,significantly higher serum levels of IL-1β,IL-6,and TNF-αand more macrophages in the uterine tissue,and activation of the TLR4/NF-κB signaling pathway.The 7 d phenol and low-concentration bacterial solution models were judged to be mild pelvic inflammatory disease models,and the

关 键 词：非手术法 盆腔炎 动物模型 子宫病理 

分 类 号：R-33[医药卫生]