摘要目的:探讨皂公二藤汤（Zaogong Erteng decoction，ZGETD）治疗子宫内膜炎的可能作用机制。方法:利用子宫角注射2.5 mg/mL脂多糖诱导建立子宫内膜炎模型，给予低剂量ZGETD、高剂量ZGETD和阿莫西林治疗7 d。7 d后观察小鼠子宫的外观和子宫组织的病理变化，计算小鼠的子宫指数；酶联免疫吸附试验法检测小鼠子宫组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素（interleukin，IL）1β和IL-6的表达；氧化还原反应测定小鼠子宫组织中髓过氧化物酶（myeloperoxidase，MPO）活性；网络药理学分析ZGETD的活性成分和治疗子宫内膜炎的作用靶点和信号通路；Western blotting和qRT-PCR分别检测小鼠子宫中Toll样受体4（Toll-like receptor 4，TLR4）、P65、p-P65、干扰素调节因子3（interferon regulatory factor 3，IRF3）和p-IRF3蛋白以及趋化因子 CXCL5和 CXCL8的表达；脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测子宫内膜细胞凋亡，并测定子宫内膜厚度；将治疗后的雌鼠与雄鼠交配，统计交配率、妊娠率和妊娠第8天注射侧子宫角的着床点数。 结果:ZGETD和阿莫西林对子宫内膜炎均有治疗效果，但高剂量ZGETD较低剂量ZGETD和阿莫西林，更明显地缓解了子宫组织的水肿和充血，显著降低了子宫指数（均 P=0.001），使得治疗后的小鼠子宫腔上皮平滑、完整，子宫腺体结构正常，未见出血区域和炎性细胞聚集现象；高剂量ZGETD较阿莫西林显著降低了炎性因子（TNF-α、IL-1β和IL-6）的表达和MPO活性（均 P<0.001），且治疗后趋化因子（ CXCL5和 CXCL8）的表达也显著降低（均 P<0.05）；网络药理学筛选出ZGETD治疗子宫内膜炎的相关信号通路TLR4、核因子κB（nuclear factor kappa-B，NF-κB）和TNF-α，并对其作用靶点（TLR4、NF-κB和IRF3）进行了验证，发现槲皮素、漆黄素和木犀草素是作用这些靶点最多的活性成分；高剂量ZGETD显著抑制了TLR4/NF-κB和TLR4/IRF3信号通路的激活（ P<0.05），降低了子宫内膜细胞凋亡（ P<0.05），提高了治疗后子宫内膜厚度（ P<0.001）、交配率（ P<0.001）、妊娠率（ P<0.001）和LPS注射侧子宫角的着床点数（ P=0.001）。 结论:高剂量ZGETD对子宫内膜炎具有显著的治疗效果，这与下调TLR4信号通路密切相关。

abstractsObjective:To investigate the possible mechanism of Zaogong Erteng decoction (ZGETD) in the treatment of endometritis.Methods:Femal mice were injected 2.5 mg/mL lipopolysaccharide into uterine horn to induce endometritis model. After modelling, low-dose ZGETD, high-dose ZGETD or amoxicillin was given once a day for 7 d. The appearance of the uterus and pathological changes of uterine tissue were observed 7 d later, and the uterine index was calculated. The expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in mouse uterine tissue was detected by enzyme-linked immunosorbent assay. The activity of myeloperoxidase (MPO) in mouse uterine tissue was measured by redox reaction. The active ingredients of ZGETD and the target and signal pathway of treatment of endometritis were analyzed by network pharmacology. Western blotting and qRT-PCR were used to detect the expressions of Toll-like receptor 4 (TLR4), P65, p-P65, interferon regulatory factor 3 (IRF3) and p-IRF3 proteins and chemokines CXCL5 and CXCL8 in the mouse uterus, respectively. Terminal dUTP nick end labeling detected endometrial cell apoptosis and endometrial thickness was measured. After treatment, the female rats were mated with the male rats, and the mating rate, the pregnancy rate and the number of implantation sits in the injected uterine horn on day 8 of gestation were counted. Results:Both ZGETD and amoxicillin have atherapeutic effect on endometritis, but compared with low-dose ZGETD and amoxicillin, high-dose ZGETD can significantly alleviate the edema and congestion of uterine tissue and reduce the uterine index (all P=0.001). After treatment, the uterine cavity epithelium of mice was smooth and complete, the uterine gland structure was normal, and no bleeding area and inflammatory cell aggregation were observed. Compared with amoxicillin, high-dose ZGETD significantly decreased the expression of inflammatory factors (TNF-α, IL-1β and IL-6) and MPO activity (all P<0.001). The expression of chemokines ( CXCL5 and CXCL8) was significantly reduced (all P<0.05). The signaling pathways TLR4, nuclear factor kappa-B (NF-κB) and TNF related to the treatment of endometritis by ZGETD were screened by network pharmacology, and their action targets (TLR4, NF-κB and IRF3) were verified. Quercetin, fisetin and luteolin were found to be the most active ingredients acting on these targets. High-dose ZGETD significantly inhibited the activation of TLR4/NF-κB and TLR4/IRF3 pathways ( P<0.05), decreased endometrial cell apoptosis ( P<0.05), and increased endometrial thickness ( P<0.001), mating rate ( P<0.001), pregnancy rate ( P<0.001) and implantation site number of uterine horn on the injection side of LPS after treatment ( P=0.001). Conclusion:High-dose ZGETD has a significant therapeutic effect on endometritis, which may be closely related to the down-regulation of TLR4 signaling pathway.