【摘要】 目的:研究活血通降方对Barrett食管(Barrett esophagus, BE)模型大鼠PGRN及TNF-α/TNFR1/NF-κB/CDX2通路的影响，探讨其防治食管BE化生的作用机制。方法:将SD大鼠随机分为对照组、模型组、西药组及中药高、中、低剂量组，每组10只。对照组行腹腔开关术，其余各组均采用“食管-十二指肠端侧吻合+保留全胃术联合铁剂注射”方法建立BE大鼠模型。造模4周后，各组分别给予相应剂量药物或相同体积生理盐水灌胃，2次/d,连续给药18周。苏木精-伊红染色评价食管黏膜病理表现；采用酶联免疫吸附法检测血清TNF-α、IL-8含量；采用蛋白免疫印记法检测食管组织中TNF-α、TNFR1、p-P65、CDX2、PGRN、GRN、MMP-12、SLPI和IL-8蛋白表达水平。结果:对照组大鼠未发生BE化生及肠化(intestinal metaplasia, IM),模型组大鼠BE化生率为83.3%,明显高于中药治疗各组的50.0%、37.5%、28.6%以及西药组的57.1%,差异有统计学意义(P=0.023)。模型组出现IM的比例为66.7%,明显高于中药治疗各组的33.3%、12.5%、14.3%以及西药组的42.9%,差异有统计学意义(P=0.044)。其中，中药中、高剂量组BE化生率及IM发生率均最低。与对照组相比，模型组大鼠血清中TNF-α、IL-8的含量，食管组织中TNF-α、TNFR1、p-P65、CDX2、GRN、MMP-12和IL-8的表达水平均明显升高(P<0.05),PGRN和SLPI的表达均显著下降(P<0.05)。给予药物干预后，中药各剂量组大鼠TNF-α、IL-8含量，食管组织中TNF-α、TNFR1、p-P65、CDX2、GRN、MMP-12和IL-8的表达水平均显著降低(P<0.05);PGRN和SLPI的表达均上调(P<0.05)。其中，中药中、高剂量组血清中TNF-α、IL-8的含量及食管组织中GRN表达的降低均优于西药组(P<0.05)。结论:活血通降方可有效预防和治疗BE的化生及IM的发生，其作用机制可能与上调PGRN的表达及抑制炎症因子TNF-α、IL-8等的释放，进而抑制TNF-α/TNFR1/NF-κB/CDX2信号通路的活化有关。更多还原

【Abstract】 Objective: To explore the mechanism of the Huoxue Tongjiang formula in preventing and treating Barrett esophagus(BE) by investigating its effects on PGRN and the TNF-α/TNFR1/NF-κB/CDX2 pathway in rats with BE. Methods: SD rats were randomly assigned to six groups: a control group, a model group, a Western medicine group, a high-dose traditional Chinese medicine group, a medium-dose traditional Chinese medicine group, and a low-dose traditional Chinese medicine group, with 10 rats in each group. The control group underwent laparotomy, whereas the other groups established BE rat models using the "esophago-duodenal end-to-side anastomosis combined with total gastrectomy and iron injection" method. Four weeks post-modeling, each group received the corresponding dose of medication or an equivalent volume of physiological saline via gavage, administered twice daily for a continuous duration of 18 weeks. HE staining was performed to evaluate the pathological changes in the esophageal mucosa. An enzyme-linked immunosorbent assay(ELISA) was utilized to detect serum levels of TNF-α and IL-8. Western blotting was employed to assess the expression levels of TNF-α, TNFR1, p-P65, CDX2, PGRN, GRN, MMP-12, SLPI, and IL-8 proteins in the esophageal tissues. Results: No BE metaplasia or intestinal metaplasia(IM) was observed in the control group. In contrast, the BE metaplasia rate in the model group was 83.3%, significantly higher than the rates of 50.0%, 37.5%, and 28.6% in the traditional Chinese medicine treatment groups, as well as 57.1% in the Western medicine group(P=0.023). The incidence of IM in the model group was 66.7%, significantly higher than the incidences of 33.3%, 12.5%, and 14.3% in the traditional Chinese medicine treatment groups and 42.9% in the Western medicine group(P=0.044). Among these groups, the BE metaplasia rate and IM incidence were lowest in the medium and high-dose traditional Chinese medicine groups. Compared to the control group, serum levels of TNF-α and IL-8 in the model group, along with expression levels of TNF-α, TNFR1, p-P65, CDX2, GRN, MMP-12, and IL-8 in esophageal tissues, were significantly elevated(P<0.05). In contrast, the expressions of PGRN and SLPI were significantly decreased(P<0.05). Following drug intervention, levels of TNF-α and IL-8 in the serum of rats in each traditional Chinese medicine dosage group, along with expression levels of TNF-α, TNFR1, p-P65, CDX2, GRN, MMP-12, and IL-8 in esophageal tissues, were significantly reduced(P<0.05). Conversely, the expressions of PGRN and SLPI were upregulated(P<0.05). Among these groups, the medium and high-dose traditional Chinese medicine groups demonstrated superior effects on reducing serum TNF-α and IL-8 levels, as well as on the expression of GRN in esophageal tissues compared to the Western medicine group(P<0.05). Conclusion: The Huoxue Tongjiang formula effectively prevents and treats BE metaplasia and IM. Its mechanism may involve up-regulating the expression of PGRN and inhibiting the release of inflammatory factors TNF-α and IL-8, thereby inhibiting the activation of the TNF-α/TNFR1/NF-κB/CDX2 signaling pathway.更多还原