Investigating the Anti-Inflammatory and Anti-Fibrotic Mechanism of Yangjing Tongluo Formula (养精通络方) in Uterine Adhesions via the NF-κB Signaling Pathway

Author：WANG Yonglian1, ZENG Jing2, ZHOU Fang1, CHEN Jiaying1, LIU Muya1

Unit：1.The First Clinical College of Chinese Medicine, Hunan University of Chinese Medicine, Changsha Hunan 410208, China; 2.The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha Hunan 410007, China

Quote：引用：王永连，曾晶，周芳，陈嘉盈，刘牧涯.基于NF-κB信号通路探讨养精通络方抗宫腔粘连炎症及纤维化的作用机制[J].中医药导报,2026,32(2):7-13.

DOI：10.13862/j.cn43-1446/r.2026.02.002

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Abstract：

Objective: To investigate the mechanism of Yangjing Tongluo formula (YJTL) against endometrial inflammation and fibrosis in rats with intrauterine adhesions (IUA). Methods: Totally 40 SPF-grade SD rats were randomly divided into a normal group (n=8) and a modeling group (n=32). An IUA rat model was established using a dual-injury method combining mechanical endometrial curettage with bacterial infection. Successfully modeled rats were randomly allocated into model group, Progynova group [0.206 mg/(kg·d)], YJTL low-dose group [6.43 mg/(kg·d)], and YJTL high-dose group [12.86 mg/(kg·d)], with 8 rats in each group. Both the normal and model groups received an equal volume of normal saline via gavage. After two weeks of administration, samples were collected. Uterine morphology was observed and the uterine index was calculated. Histopathological changes were assessed by hematoxylin-eosin (HE) staining. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of nuclear factor-κB (NF-κB), p65, and phosphorylated p65 (p-p65) in uterine tissues were analyzed by Western blotting. The expression levels of p65 and phosphorylated p65 (p-p65) in uterine tissues were analyzed by immunofluorescence. Results: (1) Gross observation and uterine index: Compared with the normal group, the model group exhibited marked uterine congestion and edema, along with a significant increase in the uterine index (P<0.01). Compared with the model group, the YJTL groups showed obvious alleviation of uterine edema and a significant reduction in the uterine index (P<0.05). (2) HE staining: Compared with the normal group, the model group displayed loose endometrial stroma, sparse glandular arrangement, and extensive infiltration of granulocytes and lymphocytes. Compared with the model group, the high-dose YJTL group showed significantly thickened endometrium, increased number of glands, and markedly reduced granulocytes and lymphocytes. (3) ELISA results: Serum levels of TNF-α and IL-1β were significantly elevated in the model group compared with the normal group (P<0.01). Serum levels of TNF-α and IL-1β were significantly reduced in the YJTL-treated groups compared with the model group (P<0.01). (4) Western blotting and immunofluorescence results: YJTL administration at various doses inhibited the expression of NF-κB, p65, and p-p65 proteins to different extents (P<0.05). Conclusion: Yangjing Tongluo formula can effectively inhibit the activity of NF-κB, p65, and p-p65 proteins, thereby exerting anti-inflammatory and anti-fibrotic effects. Its mechanism of action is likely closely associated with the effective regulation of the NF-κB signaling pathway.

Key words：intrauterine adhesions; Yangjing Tongluo formula; NF-κB signaling pathway; inflammatory factors; fibrosis; rats

摘要：

目的：探讨养精通络方（YJTL）抗宫腔粘连大鼠子宫内膜炎症及纤维化的作用机制。方法：40只SPF级SD大鼠随机分为正常组（n=8）和造模组（n=32）。通过机械性子宫内膜刮除联合细菌感染的双重损伤法构建宫腔粘连大鼠动物模型。将造模成功大鼠随机分成模型组、补佳乐组[0.206 mg/（kg·d）]、YJTL低量组[6.43 mg/（kg·d）]及YJTL高量组[12.86 mg/（kg·d）]，每组8只。正常组与模型组均予以等体积生理盐水灌胃。各组给药2周后取材待用。对子宫组织进行形态学观察并计算子宫指数；采用苏木精-伊红（HE）染色评估子宫组织病理学改变；通过酶联免疫吸附试验（ELISA）检测大鼠血清肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的水平；运用蛋白质印迹（Western blotting）法分析子宫组织中核转录因子-κB（NF-κB）、p65及其磷酸化形式（p-p65）的表达水平；免疫荧光法检测p65及p-p65蛋白表达。结果：（1）大鼠子宫组织肉眼观及子宫指数：与正常组比较，模型组大鼠子宫充血水肿，子宫指数明显上升（P<0.01）；与模型组比较，YJTL组子宫组织水肿明显消退，子宫指数显著降低（P<0.05）。（2）HE染色子宫组织病理学图片示：与正常组比较，模型组大鼠内膜间质结构疏松，腺体排列稀疏，大量粒细胞与淋巴细胞浸润；与模型组比较，YJTL高量组大鼠内膜明显增厚，腺体数量增多，粒细胞与淋巴细胞显著减少。（3）ELISA检测结果显示：与正常组比较，模型组大鼠TNF-α、IL-1β水平显著升高（P<0.01）；与模型组比较，YJTL给药组大鼠TNF-α、IL-1β水平显著降低（P<0.01）。（4）Western blotting法及免疫荧光结果显示：YJTL各剂量组可不同程度地抑制NF-κB、p65及p-p65蛋白表达（P<0.05）。结论：养精通络方能够有效抑制NF-κB、p65及p-p65蛋白活性，从而发挥抗炎、抗纤维化作用，其作用机制可能与有效调控NF-κB信号通路密切相关。

关键词：宫腔粘连；养精通络方；NF-κB信号通路；炎症因子；纤维化；大鼠

Release time：2026-03-05

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