特发性肉芽肿性乳腺炎大鼠模型建立及IL-6、IL-10免疫失衡探讨

Establishment of idiopathic granulomatous mastitis rat model and discussion on IL-6,IL-10 immune imbalance

目的 建立特发性肉芽肿性乳腺炎(idiopathic granulomatous mastitis, IGM)动物模型,并与急性乳腺炎动物模型区分IL-6、IL-10免疫失衡状态。方法 取刚哺乳完的健康成年雌性Wistar大鼠、裸鼠,胃复安注射建立高泌乳素血症状态,随机分为空白对照组A、IGM病变组织匀浆+弗氏完全佐剂混悬液组B、弗氏完全佐剂组C、急性乳腺炎脓液组D,以注射的方式诱导建立IGM动物模型,HE染色评价成模情况,免疫组化法检测IGM与急性乳腺炎动物模型IL-6、IL-10的表达差异。结果 (1)裸鼠组第1~14天均未出现乳房部位红、肿等炎症反应;(2)A、C组大鼠均未出现乳房部位炎症反应,B、D组大鼠第3天开始出现炎症包块反应,且情绪烦躁、饮食减少,至第14天B、D组所有大鼠出现乳腺炎症包块反应;(3)病理组织切片HE染色显示:A、C组大鼠未见明显炎症细胞等浸润;B组大鼠乳腺小叶和乳导管周围可见大量淋巴细胞、中性粒细胞及少量浆细胞等炎性细胞浸润,与人IGM患者病变类似;D组可见脓肿坏死和大量中性粒细胞浸润;(4)免疫组化结果显示B组IL-6异常升高,IL-10降低,D组IL-6、IL-10均升高。结论 经产哺乳后大鼠是建立IGM模型的理想动物,IGM病变组织匀浆+弗氏完全佐剂混悬液向乳房注射可有效建立IGM动物模型;裸鼠因其自身缺乏免疫力,无法成模,说明IGM与自身免疫反应有关;与急性乳腺炎模型比较,IGM造模成功组存在IL-6、IL-10免疫失衡状态。

Objective To set up an animal model for idiopathic granulomatous mastitis(IGM),and to distinguish the immune imbalance status of IL-6 and IL-10 from acute mastitis animal model.Methods Healthy adult female Wistar rats and nude mice just completing lactation were selected and injected with metoclopramide to set up the hyperprolactinemia state, and the rats were randomly divided into the A blank control group.B IGM lesion tissue homogenate+Freund′s complete adjuvant suspension group;C Freund complete adjuvant group;D acute mastitis & pus group, IGM animal model was induced by injection.Pathological HE staining was used to evaluate the model formation state, and the expression differences of IL-6 and IL-10 between IGM and acute mastitis animal models were detected by immunohistochemistry.Results(1)No inflammatory reactions involving the redness and swelling in the breast area were found in nude mice during days thoroughly from the 1 st Day to 14 th Day.(2)No inflammatory reaction occurred in breast sites of mice in group A and C,and inflammatory mass reaction began to occur in group B and D on the 3 rd Day.Inflammatory rats were agitated, and the appetite was decreased.All the rats in group B and D showed mammary inflammatory mass reactions on the 14 th Day.(3)HE stainings of rat pathological tissues showed no significant infiltrations involving inflammatory cells in group A and C.Abundant lymphocytes and neutrophils, few plasma cells and other cells were noted to have inflammatory cell infiltrations around the breast lobules and milk duct in group B,which were similar to the lesions of patients with IGM.Abundant abscess necrosis and neutrophil infiltration were observed in group D.(4)Immunohistochemical results showed that IL-6 was abnormally elevated and IL-10 was lowered in group B,while both IL-6 and IL-10 were elevated in group D.Conclusion The postpartum & post-lactation rats are ideal animals to set up IGM model, which is supported to be effectively by IGM pathological tissue homogenate+breast injection with the suspension of Freund′s complete adjuvant.The nude mice can′t be modeled due to their lack of autoimmunity, indicating that IGM is related to autoimmune response.Compared with acute suppurative mastitis model, IL-6 and IL-10 immune imbalance is found in the successful modeling IGM group.