文章摘要
张双艺,祝琦,许雷来,等.槐耳清膏治疗大鼠肉芽肿性小叶性乳腺炎模型的作用研究[J].浙江中医药大学学报,2024,48(11):1345-1354.
槐耳清膏治疗大鼠肉芽肿性小叶性乳腺炎模型的作用研究
Study on the Effect of Huaier Water Extracts in Treating the Rat Model of Granulomatous Lobular Mastitis
DOI:10.16466/j.issn1005-5509.2024.11.003
中文关键词: 免疫失衡  肉芽肿性小叶性乳腺炎  槐耳  槐耳清膏  肉芽肿性小叶性乳腺炎动物模型  肠道菌群  炎症
英文关键词: immune imbalance  granulomatous lobular mastitis  Huaier(Trametes robiniophila Murr.)  Huaier water extracts  animal model of granulomatous lobular mastitis  intestinal flora  inflammation
基金项目:浙江省中医药科技计划项目(2023ZL416)
作者单位
张双艺 浙江中医药大学第一临床医学院 杭州 310053 
祝琦 浙江中医药大学附属第一医院浙江省中医院 
许雷来 浙江中医药大学附属第一医院浙江省中医院 
谢小红 浙江中医药大学附属第一医院浙江省中医院 
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中文摘要:
      [目的] 探讨槐耳清膏通过降低炎症水平治疗肉芽肿性小叶性乳腺炎(granulomatous lobular mastitis,GLM)大鼠的作用。[方法] 将雌性SD大鼠随机分为正常组、模型组、槐耳组[槐耳清膏4.5 ?g/(kg·d)]、激素组[醋酸泼尼松龙0.0018 ?g/(kg·d)],利用GLM病变组织与弗氏完全佐剂混合后的组织匀浆诱导GLM大鼠模型。给药14 d后,对各组大鼠进行乳腺大体观察,以大鼠乳腺炎症指数评分表进行炎症指数评分,苏木精-伊红染色后进行乳腺组织病理学观察,并通过酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测大鼠血清炎症细胞因子的表达水平,采用16S rRNA扩增子测序技术测定各组大鼠肠道菌群的多样性、丰度以及肠道菌群相对丰度。[结果] 与正常组比较,模型组大鼠乳房皮色红,出现明显肿块,且乳腺炎症指数显著上升(P<0.05);病理学改变包括形成以乳腺小叶为中心的肉芽肿,伴大量淋巴细胞、浆细胞等炎症细胞浸润,证实成功诱导GLM大鼠模型。与模型组比较,槐耳组及激素组大鼠乳房外观红肿改善,肿块变软,炎症指数均下降(P<0.05,P<0.05),乳腺炎症程度明显改善,炎症细胞减少,血清炎症因子白细胞介素-6(interleukin-6,IL-6)、白细胞介素-17A(interleukin-17A,IL-17A)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平显著降低(P<0.05,P<0.05,P<0.05),乳腺组织淋巴细胞、浆细胞等炎症细胞浸润减少。模型大鼠肠道菌群发生明显变化,差异具有统计学意义(P<0.05)。模型组大鼠肠道菌群中乳杆菌目、放线菌门、双歧杆菌目、红蝽菌纲丰度下调,柄杆菌目和脱硫菌门丰度上调。槐耳清膏干预后,模型大鼠肠道菌群中梭菌纲丰度下调,拟杆菌门、红螺菌目丰度上调。[结论] 槐耳清膏能够缓解GLM模型大鼠炎症症状,有效减轻GLM模型大鼠乳腺组织炎症细胞浸润,抑制上皮细胞增生及炎性肉芽肿的形成,降低GLM模型大鼠血清炎症细胞因子水平,同时调节大鼠肠道菌群,其作用机制可能跟增强大鼠自身免疫力、调节肠道菌群,进而有效缓解炎症反应有关。
英文摘要:
      [Objective] To explore the therapeutic effect of Huaier(Trametes robiniophila Murr.) water extracts on granulomatous lobular mastitis(GLM) rat by reducing inflammation levels. [Methods] Female SD rats were randomly divided into normal group, model group, Huaier group[Huaier water extracts 4.5 ?g/(kg·d)] and hormone group[prednisone acetate 0.0018 ?g/(kg·d)]. The GLM model was induced with tissue homogenate mixture of GLM tissue and Freund’s complete adjuvant. After 14 days of administration, the mammary glands of each group were observed; the inflammation index score of the rat mammary gland was evaluated by breast inflammation index score; pathological observation of breast tissue was performed after hematoxylin-eosin(HE) staining and the level of inflammatory factors in the rat serum was detected by enzyme-linked immunosorbent assay(ELISA) method; 16S rRNA amplicon sequencing technique was used to determine the diversity and richness of intestinal flora, and the relative abundance of intestinal flora in each group. [Results] Compared with normal group, the rats in model group showed symptoms such as the red breast skin, obvious lumps and significantly increased breast inflammation index score(P<0.05). Pathological changes included the formation of granulomas centered on the breast lobules, with a large number of lymphocytes, plasma cells and other inflammatory cells infiltration, which confirmed the successful induction of GLM rat model. Compared with model group, the breast swelling was improved, and the breast inflammation index score of Huaier group and hormone group both decreased(P<0.05, P<0.05). And the level of inflammatory factors interleukin-6(IL-6), interleukin-17A(IL-17A) and tumor necrosis factor-α(TNF-α) in serum significantly decreased(P<0.05, P<0.05, P<0.05). It could alleviate the infiltration of inflammatory cells in the mammary gland tissue of model group. Significant alterations in the gut microbiota of GLM rat models were observed, with statistically significant differences(P<0.05). After modeling, the abundances of Lactobacillales, Actinobacteria, Bifidobacteriales and Coriobacteriia were downregulated, while those of Caulobacterales and Desulfobacterota were upregulated within the gut microbiota. After the intervention with Huaier water extracts, the abundances of Clostridia were downregulated, while those of Bacteroidetes and Rhodospirillales were upregulated in the gut microbiota of GLM model rats. [Conclusion] Huaier water extracts could alleviate the infiltration of inflammatory cells in the mammary gland tissue of GLM model rats; reduce the proliferation of epithelial cells and the formation of inflammatory granuloma; decrease the level of inflammatory cytokines in the serum of GLM rat model and adjust the intestinal flora of GLM rat models, which may be related to its ability to improve the immunity and relieve the inflammatory response.
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