【摘要】 目的：慢性输卵管炎是导致女性继发性不孕、输卵管妊娠及慢性盆腔疼痛的重要诱因，其病理特征主要表现为输卵管黏膜粘连、纤毛运动功能障碍及管腔闭锁。由于临床研究受限于人体活检组织获取的困难，建立一种稳定、标准且符合人体病理演变过程的动物模型对探讨慢性输卵管炎发病机制及筛选治疗药物具有重要的学术价值。目前尚缺乏统一明确、成功率高且经济实用的慢性输卵管炎动物模型。本研究旨在评估鼠衣原体（Chlamydia muridarum,CM）与混合菌（mixed bacteria,MB）诱导SD大鼠慢性输卵管炎的效果，并筛选优化MB造模的最佳参数。方法：本研究分为病原体模型比较和MB剂量效应评估2个阶段。病原体模型比较：选取30只SPF级雌性SD大鼠，随机分为对照（control,CON）组、CM组[接种浓度2×10⁷包涵体形成单位（inclusion forming units,IFU）/mL]和MB组[接种浓度3×10⁹菌落形成单位（colony forming units,CFU）/mL]。采用生殖道接种法造模，采用苏木精-伊红（hematoxylin and eosin,HE）染色及透射电子显微镜（transmission electron microscopy,TEM）分别于造模后第1、7、14、21、28天观察输卵管组织病理形态及上皮纤毛、线粒体等超微结构变化。MB剂量效应评估：选取30只无特定病原体级雌性SD大鼠分为正常组、低浓度MB组(3×10¹⁰ CFU/mL)及高浓度MB组(3×10¹¹ CFU/mL)。采用酶联免疫吸附测定（enzyme-linked immunosorbent assay,ELISA）技术检测造模后第7、14天血清肿瘤坏死因子-α（tumor necrosis factor-alpha,TNF-α）水平，结合显微镜下的组织形态学评分对炎症程度进行客观的量化评估。结果：CM组与MB组均成功诱发了典型的慢性输卵管炎表现，光镜下可见输卵管间质水肿、管壁增厚及明显的淋巴细胞和浆细胞浸润。在TEM超微结构下，造模组大鼠表现出极具特征性的细胞器损伤，包括纤毛排列紊乱、大面积倒伏脱落，轴丝结构不清晰，以及细胞质内线粒体显著肿胀、嵴断裂消失甚至出现空泡样变。时效性对比显示，在造模后第7天，MB组表现出比CM组更显著的炎症反应，其特征为中性粒细胞浸润明显增多。剂量效应分析显示，在造模后第14天，高浓度MB组大鼠血清TNF-α水平显著升高（P<0.01），且输卵管组织损伤程度加剧，呈现出典型的慢性炎症改变。结论：CM模型和MB模型均能有效模拟人类慢性输卵管炎从急性炎症反应向慢性纤维化演变的完整病理过程，且2种模型均具有较高的成功率。然而CM模型的病原体培养条件苛刻，购买及维护成本较高，实验周期较长。相比之下，MB模型所采用的混合菌菌株获取便捷、制备成本低廉，且在3×10¹¹ CFU/mL这一特定浓度下构建的MB方案，不仅能够稳定诱导大鼠产生符合慢性炎症标准的形态学改变，而且其炎症因子的表达具有良好的可量化性与可重复性，为慢性输卵管炎的基础研究、临床药物筛选及转化医学研究提供了具有重要学术价值的标准实验载体。更多还原

【Abstract】 Objective: Chronic salpingitis is an important cause of secondary infertility, tubal pregnancy, and chronic pelvic pain in women. Its pathological characteristics mainly include adhesion of the tubal mucosa, impaired ciliary motility, and luminal occlusion. Due to the difficulty in obtaining human biopsy tissues, establishing a stable, standardized animal model consistent with the pathological progression in humans is of great academic significance for investigating the pathogenesis of chronic salpingitis and screening therapeutic drugs. Currently, there is still a lack of unified, reliable, and cost-effective animal model for chronic salpingitis. This study aims to evaluate the effects of Chlamydia muridarum（CM） and mixed bacteria（MB） in inducing chronic salpingitis in SD rats and to optimize the parameters for MB modeling.Methods: The study consisted of 2 stages: pathogen model comparison and MB dose-effect evaluation. For pathogen model comparison, 30 SPF female SD rats were randomly divided into a control（CON） group, a CM group [inoculation concentration 2×107 inclusion forming units（IFU）/mL], and an MB group [inoculation concentration 3×10⁹ colony forming units（CFU）/mL]. A reproductive tract inoculation method was used to establish the model. Hematoxylin-eosin（HE） staining and transmission electron microscopy（TEM） were performed on days 1, 7, 14, 21, and 28 after modeling to observe pathological morphology of the fallopian tube and ultrastructural changes in epithelial cilia and mitochondria. For MB dose-effect evaluation, 30 specific pathogen free female SD rats were divided into a normal group, a low-dose MB group(3×10¹⁰ CFU/mL), and a high-dose MB group(3×10¹¹ CFU/m L). Serum tumor necrosis factor-α（TNF-α） levels were measured on days 7 and 14 after modeling using enzyme-linked immunosorbent assay（ELISA）. Combined with histomorphological scoring under microscopy, inflammation severity was quantitatively evaluated.Results: Both the CM group and MB group successfully induced typical manifestations of chronic salpingitis. Light microscopy showed interstitial edema of the fallopian tube, thickening of the tubal wall, and marked infiltration of lymphocytes and plasma cells. Under TEM, rats in the modeling groups exhibited characteristic organelle damage, including disordered ciliary arrangement, extensive ciliary collapse and shedding, indistinct axoneme structure, and significant mitochondrial swelling with cristae disruption or disappearance and vacuolar degeneration in the cytoplasm. Time-course comparison showed that on day 7 after modeling, the MB group exhibited a more pronounced inflammatory response than the CM group, characterized by increased neutrophil infiltration. Dose-effect analysis demonstrated that on day 14 after modeling, serum TNF-α levels were significantly elevated in the high-dose MB group（P<0.01）, accompanied by aggravated fallopian tube tissue damage with typical chronic inflammatory changes.Conclusion: Both CM and MB models can effectively simulate the complete pathological process of chronic salpingitis in humans, from acute inflammatory response to chronic fibrosis, with relatively high success rates. However, the CM model requires stringent pathogen culture conditions, higher purchase and maintenance costs, and a longer experimental cycle. In contrast, the MB model uses mixed bacterial strains that are easier to obtain and less costly to prepare. This study confirmed that the MB protocol constructed at a concentration of 3×10¹¹ CFU/mL can stably induce morphological changes consistent with chronic inflammation in rats, and the expression of inflammatory factors shows good quantifiability and reproducibility. This model provides a standardized experimental platform with important academic value for basic research, clinical drug screening, and translational studies of chronic salpingitis.更多还原